supplement i to the japanese pharmacopoeia fourteenth edition

The Ministry of Health, Labour andWelfare Ministerial Notification No. 395

In accordance with the provision of Paragraph 1 of Article 41 of the Pharmaceuti-cal Affairs Law (Low No. 145, 1960), we hereby revise a part of the Japanese Phar-macopoeia (Ministerial Notification No. 111, 2001) as follows, and the revisedJapanese Pharmacopoeia shall come into effect on January 1, 2003. The drugs in-cluded in the Japanese Pharmacopoeia (hereinafter referred to as ``the old JapanesePharmacopoeia'') [limited to those included in the Japanese Pharmacopoeia whosestandards are changed in accordance with this notification (hereinafter referred to as``the new Japanese Pharmacopoeia'')] and the approval for which is given on Janu-ary 1, 2003 as stipulated in Paragraph 1 of Article 14 (including cases where it shallapply mutatis mutandis under Article 23 of the Law; the same hereinafter) [includingthe drugs designated as those exempted from manufacturing or import approval(Ministerial Notification on the designation of the drugs exempted from manufactur-ing or import approval in accordance with the provision of Paragraph 1 of Article 14of the Law) (hereinafter referred to as ``the drugs exempted from approval'')] can betreated, up to June 30, 2004, in such a way that the standards established by the oldJapanese Pharmacopoeia (limited to the contents concerned with the relevant drugs)may be accepted as those established by the new Japanese Pharmacopoeia. The drugsincluded in the new Japanese Pharmacopoeia (excluding those in the old JapanesePharmacopoeia) the approval for which is given on January 1, 2003, as stipulated inParagraph 1 of Article 14 of the Law (including the drugs exempted from approval),can be treated, up to June 30, 2004, as those not included in the new Japanese Phar-macopoeia.

Chikara SakaguchiThe Minister of Health, Labour and Welfare

December 27, 2002

(The texts referred to by the term ``as follows'' are omitted here. All of them aremade available for public exhibition at the Evaluation and Licensing Division, Phar-maceutical and Medical Safety Bureau, Ministry of Health, Labour and Welfare, ateach Regional Bureau of Health and Welfare and at each Prefectural Office inJapan.)

The term ``as follows'' here indicates the contents from Part I to Ultraviolet-visible ReferenceSpectra in the Supplement I to the Japanese Pharmacopoeia Fourteenth Edition(pp.1359 – 1610).

CONTENTS

Preface ...................................................... iSupplement I to The Japanese Pharmacopoeia,Fourteenth Edition, Part I................. 1359–1540

General Notices ................................... 1359General Rules for Preparations ............... 1361General Tests, Processes and Apparatus ... 136323. Infrared Spectrophotometry.............. 136352. Residue on Ignition Test .................. 136366. Vitamin A Assay ............................ 136470. Reference Standards; Reagents, Test Solutions;

Standard Solutions for Volumetric Analysis;Standard Solutions; Matching Fluids for Color;Optical Filters for Wavelength and Transmis-sion Rate Calibration; and Measuring Instru-ments, Appliances ............................ 1365(1) Reference Standards................. 1365(2) Reagents, Test Solutions ........... 1365(3) Standard Solutions for Volumetric

Analysis................................. 137372. Conductivity Measurement................ 137373. Determination of Bulk and Tapped

Densities ....................................... 1375Official Monographs............................. 1377

Supplement I to The Japanese Pharmacopoeia,Fourteenth Edition, Part II................ 1541–1566

General Rules for Crude Drugs ............... 1541Official Monographs............................. 1543

Infrared Reference Spectra ................ 1567–1586Part I ................................................ 1567

Ultraviolet-visible Reference Spectra .... 1587–1610Part I ................................................ 1587

General Information1. Aristolochic Acid............................ 16115. International Harmonization Implemented

in the Japanese Pharmacopoeia FourteenthEdition ......................................... 1611

12. Preservatives-Effectiveness Tests ........ 161617. Basic Requirements for Viral Safety of

Biotechnological/Biological Productslisted in Japanese Pharmacopoeia ...... 1618

18. Qualification of Animals as Origin ofAnimal-derived Medicinal Productsprovided in the General Notices 39 ofJapanese Pharmacopoeia and OtherStandards...................................... 1631

19. SDS-Polyacrylamide GelElectrophoresis............................... 1634

Appendix .............................................. 1641

Index.................................................... 1643Index in Japanese.................................... 1659

ii

Preface

The Fourteenth Edition of the Japanese Phar-macopoeia was promulgated on March 30, 2001 byMinisterial Notiˆcation No. 111 of the Ministry ofHealth, Labour and Welfare. To keep pace withprogress in medical and pharmaceutical sciences, inNovember 2001, the Council, at a meeting of theCommittee on Japanese Pharmacopoeia (JP), estab-lished the basic principles for the preparation of theJP Fifteenth Edition, setting out the characteristicsand roles of the JP, the deˆnite measures for the revi-sion, the date of the revision, and the organization ofthe Subcommittee on JP.

At the above meeting, the following ``ˆve pillars''were established as the basic principles of the JP:Making it more substantial by including all drugswhich are important from the viewpoint of health careand medical treatment; Making prompt partial revi-sion as necessary and facilitating smooth administra-tive operation; Promoting international harmoniza-tion; Ensuring transparency regarding the revision anddissemination to the public of the JP; and Positivelyintroducing contemporary analytical tests and de-veloping reference standards. It was decided at themeeting that each panel set up under the Subcommit-tee on JP should make eŠorts, on the basis of theseprinciples, to ensure that the JP is used more eŠective-ly in the ˆelds of health care and medical treatment bytaking appropriate measures, including getting the un-derstanding and cooperation of other parties con-cerned.

The JP should comprise an o‹cial standard beingrequired to assure the quality of drugs in this countryin response to the progress in science and technologyand clinical demands at the time, it should deˆne thestandards for speciˆcations as well as the methods oftests to assure the overall quality of all drugs in princi-ple, and it should have a role in clarifying the criteriafor quality of drugs which are recognized to be im-portant from the viewpoint of medical treatment.

At the same time, it was agreed that the JP shouldbe prepared with the aid of the knowledge and ex-perience of many persons involved in the pharmaceuti-cals, that it should have the characteristics of an o‹-cial standard, which might be widely used by all par-ties concerned, that it should provide information andunderstanding about the quality of drugs to the pub-lic, and that it should be conducive to smooth andeŠective government control of the quality of drugs,

and to securing and maintaining international con-sistency.

It was also agreed that JP articles should coverdrugs which are important from the viewpoint ofhealth care and medical treatment, clinical results andfrequency of use, as soon as possible after they reachthe market.

It was also decided to make a deˆnite rule for selec-tion of articles, by clarifying the signiˆcance of, andstandards for selection. The JP Fifteenth Edition wasdecided to be slated for completion in April 2006.

Under the Subcommittee on JP, the followingtwelve panels and two provisional panels were estab-lished at ˆrst: Panel on Planning and Revisions; Panelon the Selection of Articles; Panel on Nomenclature;Panel on Excipients; First Panel on Medicinal Chemi-cals; Second Panel on Medicinal Chemicals; Panel onBiologically Derived Drugs; Panel on Biological Tests;Panel on Physico-chemical Tests; Panel on MaterialSciences; Panel on Preparations; Panel on CrudeDrugs; Provisional First Panel on Antibiotics;Provisional First Panel on Crude Drugs. Some of thenames of the above panels were changed as follows,due to the reorganization of the PharmaceuticalAŠairs and Food Sanitation Council (PAFSC) inNovember 2001: Panel on Nomenclature to Panel onNomenclature for Pharmaceutical Chemicals; FirstPanel on Medicinal Chemicals and Second Panel onMedicinal Chemicals were integrated into Panel onMedicinal Chemicals; Panel on Physico-chemicalTests, Panel on Material Sciences and Panel on Prepa-rations were integrated into Panel on Physico-chemi-cal Tests; Panel on Crude Drugs and Provisional FirstPanel on Crude Drugs were integrated into Panel onCrude Drugs; Provisional First Panel on Antibioticsto Panel on Antibiotics. Under the Panel on Planningand Revisions, the following two panels were newly es-tablished: Panel on General Revisions and Panel onPharmacopoeial Harmonization (PDG).

In the Committee on Japanese Pharmacopoeia,Tadao Terao took the role of chairman from Novem-ber 1997 to December 2000 and Mitsuru Uchiyamafrom January 2001 to December 2002.

With the reform of central government ministriesand agencies in January 2001, the Ministry of Healthand Welfare became the Ministry of Health, Labourand Welfare, and the Committee on Japanese Phar-macopoeia (CJP) came under the authority of the

iiii Supplement I, JP XIVPreface

Minister of Health, Labour and Welfare. At the sametime, the Central Pharmaceutical AŠairs Councilbecame the Pharmaceutical AŠairs and Food Sanita-tion Council (PAFSC) and Mitsuru Uchiyama wasnominated as chairman of the CJP.

It was decided that the JP will be revised not onlyevery ˆve years, in line with the basic principles for thepreparation of the JP Fifteenth Edition, but also asnecessary to take account of recent progress of scienceand in the interests of international harmonization. Inaccordance with the revision principles, the panelscontinued discussions on selection of articles, and re-visions for General Notices, General Rules for Prepa-rations, General Tests, and monographs on drugs.

Among the items discussed, the following two revi-sion drafts were separately examined by the Commit-tee on JP in November 2001, followed by PAFSC inDecember 2001, and then submitted to the Minister ofHealth, Labour and Welfare, and Ministerial Notiˆca-tion No.151 promulgated these revisions on March 29,2002: Addition of a paragraph ``In principle, animalsused as a source of materials for preparing phar-maceutical drugs must be healthy.'' to General No-tices; and Deletion of a monograph ``Phenacetin''.

Draft revisions covering subjects in General No-tices, General Rules for Preparations, General Rulesfor Crude Drugs, General Tests, and monographs ondrugs, for which discussions were ˆnished betweenJune 2000 and February 2001, were prepared for asupplement to the book. They were examined by theCommittee on JP in September 2002, followed byPAFSC in December 2002, and then submitted to theMinister of Health, Labour and Welfare.

Numbers of discussions in the panels to preparesupplement drafts were as follows: Panel on the Prin-ciples of Revisions, 6 times; Panel on the Selection ofArticles, 2 times; Panel on Nomenclature, 11 times;Panel on Excipients, 10 times; First Panel on Medici-nal Chemicals, 10 times; Second Panel on MedicinalChemicals, 16 times; Panel on Biologically DerivedDrugs, 8 times; Panel on Biological Tests, 9 times;Panel on Physico-chemical Tests, 8 times; Panel onMaterial Sciences, 8 times; Panel on Preparations, 5times; Panel on Crude Drugs, 6 times; ProvisionalFirst Panel on Antibiotics, 27 times; Provisional FirstPanel on Crude Drugs, 7 times. Numbers of discus-sion in the panels, which were renamed due to the re-organization of PAFSC, were as follows: Panel onNomenclature for Pharmaceutical Chemicals, 2 times;Panel on Physico-chemical Tests, 1 time; Panel onCrude Drags, 3 times; Panel on General Revisions, 1time.

It should be noted that in the preparation of the

drafts for the Supplement I, generous cooperation wasgiven by the Technical Committee of the Pharmaceuti-cal Manufacturer's Association of Tokyo and ofOsaka, the Crude Drugs Association of Tokyo, theJapan Pharmaceutical Excipients Council, the Federa-tion of Crude Drugs Associations of Japan, the JapanAntibiotics Research Association, the Japan Flavorand Fragrance Manufacturer's Association, the JapanMedical Plants Federation, the Japan PharmaceuticalManufacturer's Association, the Japanese Society ofHospital Pharmacists, the Japan Pharmaceutical As-sociation, and the Japan Oilseed Processors Associa-tion.

In consequence of this revision, the JP FourteenthEdition carries 881 articles in Part I owing to the addi-tion of 31 articles and the deletion of 8 articles; and481 articles in Part II owing to the addition of 15 arti-cles and the deletion of 3 articles.

The principles of description and the salient pointsof the revision in this volume are as follows:

1. The Supplement I to JP Fourteenth Editioncomprises the following items, in order: Notiˆcationof the Ministry of Health, Labour and Welfare; Con-tents; Preface; followed by General Notices; GeneralRules for Preparations; General Tests, Processes andApparatus; Monographs on Drugs in Part I, andGeneral Notices; General Rules for Crude Drugs;General Rules for Preparations; General Tests,Processes and Apparatus; Monographs on Drugs inPart II, then followed by Infrared Reference Spectrain Part I; Ultraviolet-visible Reference Spectra in PartI; General Information, and as appendix, the Ministryof Health, Labour and Welfare Ministerial Notiˆca-tion No. 151 (March 2002); and Cumulative Indexcontaining references to the main volume and the sup-plement I.

2. The articles in General Rules for Preparations,in General Tests, Processes and Apparatus, Mono-graphs on Drugs, Infrared Reference Spectra andUltraviolet-visible Reference Spectra are respectivelyplaced in alphabetical order.

3. The following items in each monograph are putin the order shown below, except that unnecessary i-tems are omitted depending on the nature of the drug:(1) English title(2) Commonly used name(s)(3) Latin title (only for Crude Drugs)(4) Title in Japanese(5) Structural formula or empirical formula(6) Molecular formula and molecular mass(7) Chemical name

iiiiiiSupplement I, JP XIV Preface

(8) Origin(9) Limits of the content of the ingredient(s)

and/or the unit of potency(10) Labeling requirements(11) Method of preparation(12) Description(13) Identiˆcation tests(14) Speciˆc physical and/or chemical values(15) Purity tests(16) Loss on drying, loss on ignition, and/or water(17) Residue on ignition, total ash, and/or acid-in-

soluble ash(18) Special tests(19) Isomer ratio(20) Assay or the content of the ingredient(s)(21) Containers and storage(22) Expiration date(23) Others

4. In each monograph on a drug, the followingphysical and chemical values representing the proper-ties and quality of the drug are given in the order indi-cated below, except that unnecessary items are omitteddepending on the nature of the drug:(1) Alcohol number(2) Absorbance(3) Congealing point(4) Refractive index(5) Osmolarity(6) Optical rotation(7) Viscosity(8) pH(9) Speciˆc gravity

(10) Boiling point(11) Melting point(12) Acid value(13) Saponiˆcation value(14) Ester value(15) Hydroxyl value(16) Iodine value

5. Identiˆcation tests comprise the followingitems, which are generally put in the order given be-low:(1) Coloration reactions(2) Precipitation reactions(3) Decomposition reactions(4) Derivatives(5) Visible, ultraviolet or infrared spectra(6) Special reactions(7) Cations(8) Anions

6. Purity tests comprise the following items, whichare generally put in the order given below, except that

unnecessary items are omitted depending on the na-ture of the drug:(1) Color(2) Odor(3) Clarity and/or color of solution(4) Acidity or alkalinity(5) Acid(6) Alkali(7) Chloride(8) Sulfate(9) Sulˆte

(10) Nitrate(11) Nitrite(12) Carbonate(13) Bromide(14) Iodide(15) Soluble halide(16) Thiocyanide(17) Selenium(18) Cationic salts(19) Ammonium(20) Heavy metals(21) Iron(22) Manganese(23) Chromium(24) Bismuth(25) Tin(26) Aluminum(27) Zinc(28) Cadmium(29) Mercury(30) Copper(31) Lead(32) Silver(33) Alkaline earth metals(34) Arsenic(35) Foreign matter(36) Related substances(37) Other mixtures(38) Readily carbonizable substances

7. Revisions in the General Notices are as follows:(1) In paragraph 6, an abbreviation ``lx'' for lux

was added as the main unit.(2) In paragraph 8, the deˆnition for ``a cold

place'' was revised to as ``a place having a tem-perature between 1 and 159C''.

8. Revision in the General Rules for Preparationsis as follows:(1) Injections: The description with respect to the

injection having 50 mL or more was deletedfrom section (9).

9. The following items of the General Tests,

iviv Supplement I, JP XIVPreface

Processes and Apparatus were partially revised:(1) Infrared Spectrophotometry(2) Residue on Ignition Test(3) Vitamin A Assay

10. The following tests were added to the GeneralTests, Processes and Apparatus:(1) Conductivity Measurement(2) Determination of Bulk and Tapped Densities

11. The following Reference Standard was delet-ed:

Drostanolone Propionate

12. The following Reference Standard was re-named:

Kitasamycin to Leucomycine A5

13. The following Reference Standards were ad-ded:

AceglutamideAcetylspiramycin IIAclarubicinActinomycin DArbekacin SulfateAstromicin SulfateBacitracinBekanamycin SulfateBenzylpenicillin PotassiumBenzylpenicillin SodiumBleomycin A2 HydrochlorideCarumonam SodiumCefaclorCefaloridinCefalotin SodiumCefamandole LithiumCefbuperazoneCefmenoxime HydrochlorideCefodizime SodiumCefotaximeCefotetanCefotiam Hexetil HydrochlorideCefoxitinCefpiramideCefpodoxime ProxetilCefroxadineCefteram Pivoxil MesitylenesulfonateCefuroxime AxetilChloramphenicolChloramphenicol PalmitateChloramphenicol SuccinateCiclacillinClindamycin HydrochlorideClindamycin PhosphateColistin Sulfate

Daunorubicin HydrochlorideDemethylchlortetracycline HydrochlorideDibekacin SulfateDiethanolamine FusidateDoxorubicin HydrochlorideDoxycycline HydrochlorideEnviomycin SulfateEpirubicin HydrochlorideFlomoxef TriethylammoniumFradiomycin SulfateGentamicin SulfateGramicidinGriseofulvinImipenemJosamycin PropionateKanamycin MonosulfateLatamoxef AmmoniumLenampicillin HydrochlorideLincomycin HydrochlorideLysozymeMicronomicin SulfateMitomycin COxytetracycline HydrochloridePeplomycin SulfateL-Phenethicillin PotassiumPimaricinPirarubicinPivmecillinam HydrochloridePolymixin B SulfatePuerarinPyrrolnitrinRanitidine HydrochlorideRetinol AcetateRetinol PalmitateRibostamycin SulfateRifampicinSennoside ASennoside BSiccaninSpectinomycin HydrochlorideStreptomycin SulfateSulbenicillin SodiumTalampicillin HydrochlorideTobramycinTrichomycinVancomycin Hydrochloride

14. English and Latin titles of drugs are derived,in principle, from International NonproprietaryNames (INN) for Pharmaceutical Substances recom-mended by the World Health Organization. Japanesetitles are derived from the Japanese version of thisbook. The chemical names are based on the rules ofthe International Union of Pure and Applied Chemis-

vvSupplement I, JP XIV Preface

try (IUPAC).

15. Molecular formulas of organic compounds be-gin with C and then H, followed by other involved ele-ments in the alphabetical order of the symbols of theelements.

16. Structural formulas of drugs represent, as faras possible, steric conˆgurations.

17. Test procedures in monographs in Part I are,in principle, written in full even in correspondingmonographs in Part II, and vice versa. The test proce-dures in monographs for preparations are also writtenin full even within the same part, except in the mono-graphs for preparations having a correspondingmonograph of their principal material substances.

18. In O‹cial Monographs, names of some of thereagents and the test solutions were changed to thelatest names based on the JIS.

19. The following articles were deleted from O‹-cial MonographsPart I

Drostanolone PropionateDrostanolone Propionate InjectionFloctafenineIopanoic AcidIopanoic Acid TabletsSimˆbrateSodium IopodateSodium Iopodate Capsules

Part IIPhenovalin and Magnesium Oxide PowderCompound Scopolia Extract and Tannic Acid Oint-

mentCompound Scopolia Extract and Tannic Acid Sup-

positories

20. The following articles were newly added toO‹cial Monographs:Part I

Aceglutamide AluminumBacitracinBenzylpenicillin BenzathineCefodizime SodiumCefpodoxime ProxetilChloramphenicol PalmitateChloramphenicol Sodium SuccinateClindamycin HydrochlorideColistin SulfateDaunorubicin HydrochlorideDemethylchlortetracycline HydrochlorideEnviomycin SulfateEpirubicin Hydrochloride

Erythromycin LactobionateEtizolamGramicidinKanamycin MonosulfateKetotifen FumarateKitasamycin TartrateLenampicillin HydrochlorideLysozyme HydrochlorideO‰oxacinPhenethicillin PotassiumPimaricinPirarubicinPyrrolnitrinRanitidine HydrochlorideSiccaninSodium FusidateSpectinomycin HydrochlorideTrimebutine Maleate

Part IIAlpinia O‹cinarum RhizomeAsparagus TuberBurdock FruitChrysanthemum FlowerClematis RootEucommia BarkFritillaria BulbGastrodia TuberHemp FruitJujube SeedLoquat LeafMagnolia FlowerNotopterygium RhizomePolygonum RootUncaria Thorn

21. The following monographs were revised by anaddition or a change in the Description or other items:Part I

AcetylkitasamycinAcetylspiramycinAclarubicin HydrochlorideActinomycin DAmoxicillinAmpicillinAnhydrous AmpicillinAmpicillin SodiumArbekacin SulfateAstromicin SulfateBacampicillin HydrochlorideBekanamycin SulfateBenzbromaroneBenzylpenicillin PotassiumBetamethasone Sodium Phosphate

vivi Supplement I, JP XIVPreface

Betamethasone ValerateBisacodyl SuppositoriesBleomycin HydrochlorideBleomycin SulfateCalcium Chloride InjectionCalcium Polystyrene Sulfonated-Camphordl-CamphorCarbazochrome Sodium SulfonateCarbidopaCarumonam SodiumCefaclorCefaloridinCefalotin SodiumCefamandole SodiumCefbuperazone SodiumCefmenoxime HydrochlorideCefotaxime SodiumCefotetanCefotiam Hexetil HydrochlorideCefoxitin SodiumCefpiramide SodiumCefroxadineCefteram PivoxilCeftibutenCeftriaxone SodiumCefuroxime AxetilCetraxate HydrochlorideChloramphenicolChlordiazepoxide PowderChlordiazepoxide TabletsChlormadinone AcetateChlorphenesin CarbamateCiclacillinCitric AcidAnhydrous Citric AcidClindamycin PhosphateClomifene CitrateCloxacillin Sodium1z Codeine Phosphate Powder10z Codeine Phosphate PowderCodeine Phosphate TabletsDibekacin SulfateDiclofenamideDiethylcarbamazine Citrate Tablets1z Dihydrocodeine Phosphate Powder10z Dihydrocodeine Phosphate PowderDihydroergotoxine MesilateDiltiazem HydrochlorideDipyridamoleDobutamine HydrochlorideDopamine Hydrochloride InjectionDoxorubicin Hydrochloride

Doxycycline HydrochlorideDroperidolEn‰uraneErgocalciferolErythromycinEstradiol BenzoateEstriolFamotidine for InjectionFlomoxef SodiumFlurbiprofenFolic AcidFradiomycin SulfateGentamicin SulfateGlycerinConcentrated GlycerinGriseofulvinHaloxazolamHomochlorcyclizine HydrochlorideHydrochlorothiazideHydrocortisone AcetateHydrocortisone Sodium PhosphateIdarubicin HydrochlorideIdoxuridine Ophthalmic SolutionImipenemIndometacin SuppositoriesIsoniazid InjectionIsoniazid TabletsJosamycinJosamycin PropionateKanamycin SulfateKitasamycinLatamoxef SodiumLincomycin HydrochlorideLiothyronine Sodium TabletsMagnesium Sulfate InjectionD-Mannitol InjectionMenatetrenoneMepitiostaneMethotrexateMeticraneMicronomicin SulfateMigreninMinocycline HydrochlorideMitomycin CMorphine HydrochlorideMorphine Hydrochloride InjectionMorphine Hydrochloride TabletsNorepinephrineNorepinephrine InjectionNorgestrel and Ethinylestradiol TabletsOxytetracycline HydrochloridePeplomycin SulfatePethidine Hydrochloride

viiviiSupplement I, JP XIV Preface

PhytonadionePipemidic Acid TrihydratePiperacillin SodiumPivmecillinam HydrochloridePolymixin B SulfatePotassium ClavulanatePovidone-IodinePrednisolone AcetatePrednisolone Sodium Succinate for InjectionPrimidoneProcaine Hydrochloride InjectionProcarbazine HydrochloridePyrazinamideReserpineRetinol AcetateRetinol PalmitateRibostamycin SulfateRifampicinRoxithromycinScopolamine ButylbromideSisomicin SulfateSodium Bicarbonate InjectionSodium Chloride10z Sodium Chloride InjectionStreptomycin SulfateSulbenicillin SodiumTalampicillin HydrochlorideTegafurTeicoplaninTheophyllineThiamine HydrochlorideThiamine NitrateThiopental SodiumTipepidine HibenzateTobramycinTocopherolTocopherol AcetateTriamcinoloneTrichomycinTrimetoquinol HydrochlorideVancomycin HydrochlorideXylitol Injection

Part IIAcaciaPowdered AcaciaAloePowdered AloeArecaAsiasarum RootCapsicumPowdered CapsicumCinnamon BarkPowdered Cinnamon Bark

Cod Liver OilCoptis RhizomePowdered Coptis RhizomeGardenia FruitGlycyrrhizaPowdered GlycyrrhizaImperata RhizomePowdered Japanese GentianMagnolia BarkPowdered Magnolia BarkMorphine and Atropine InjectionMulberry BarkOpium Alkaloids HydrochloridesPowdered Panax RhizomePeony RootPowdered Peony RootPhellodendron BarkPowdered Phellodendron BarkCompound Phellodendron Powder for CataplasmPhellodendron, Albumin Tannate and Bismuth

Subnitrate PowderPueraria RootRhubarbPowdered RhubarbScopolia Extract and Ethyl Aminobenzoate PowderScutellaria RootPowdered Scutellaria RootSenna LeafPowdered Senna LeafSinomenium StemSwertia HerbPowdered Swertia HerbSwertia and Sodium Bicarbonate PowderToad VenomVitamin A OilVitamin A Oil Capsules

22. The monographs, which use in Identiˆcationthe newly introduced Infrared Reference Spectra, areas follows:Part I

AcetylspiramycinAclarubicin HydrochlorideAmpicillinAnhydrous AmpicillinAmpicillin SodiumBenzylpenicillin BenzathineBenzylpenicillin PotassiumBleomycin HydrochlorideBleomycin SulfateCarumonam SodiumCefaclorCefaloridinCefalotin Sodium

viiiviii Supplement I, JP XIVPreface

Cefamandole SodiumCefmenoxime HydrochlorideCefodizime SodiumCefotaxime SodiumCefotetanCefpodoxime ProxetilCefuroxime AxetilChloramphenicolChloramphenicol Sodium SuccinateCiclacillinCitric AcidAnhydrous Citric AcidClindamycin PhosphateDemethylchlortetracycline HydrochlorideDoxorubicin HydrochlorideDoxycycline HydrochlorideErythromycinEtizolamFlomoxef SodiumGlycerinConcentrated GlycerinGriseofulvinHomochlorcyclizine HydrochlorideImipenemKetotifen FumarateKitasamycin TartrateLatamoxef SodiumLenampicillin HydrochlorideLincomycin HydrochlorideMitomycin CNorepinephrineO‰oxacinPhenethicillin PotassiumPhytonadionePivmecillinam HydrochloridePyrazinamidePyrrolnitrinRanitidine HydrochlorideRifampicinSiccaninSodium FusidateSulbenicillin SodiumTalampicillin HydrochlorideTheophyllineTrimebutine MaleateVancomycin Hydrochloride

23. The monographs, which use in Identiˆcationthe newly introduced Ultraviolet-visible ReferenceSpectra, are as follows:Part I

AcetylspiramycinAclarubicin HydrochlorideActinomycin D

BenzbromaroneBenzylpenicillin BenzathineBenzylpenicillin PotassiumBleomycin HydrochlorideBleomycin SulfateCarumonam SodiumCefaclorCefaloridinCefalotin SodiumCefamandole SodiumCefbuperazone SodiumCefmenoxime HydrochlorideCefodizime SodiumCefotaxime SodiumCefotetanCefotiam Hexetil HydrochlorideCefoxitin SodiumCefpiramide SodiumCefpodoxime ProxetilCefroxadineCefteram PivoxilCeftriaxone SodiumCefuroxime AxetilChloramphenicolChloramphenicol PalmitateChloramphenicol Sodium SuccinateCloxacillin SodiumDaunorubicin HydrochlorideDemethylchlortetracycline HydrochlorideDoxorubicin HydrochlorideEnviomycin SulfateEpirubicin HydrochlorideEtizolamFlomoxef SodiumGramicidinGriseofulvinHomochlorcyclizine HydrochlorideImipenemJosamycinJosamycin PropionateKetotifen FumarateKitasamycin TartrateLatamoxef SodiumLysozyme HydrochlorideMinocycline HydrochlorideMitomycin CNorepinephrineO‰oxacinOxytetracycline HydrochloridePeplomycin SulfatePhenethicillin PotassiumPimaricinPirarubicin

ixixSupplement I, JP XIV Preface

Potassium ClavulanatePyrrolnitrinRanitidine HydrochlorideRifampicinSiccaninTheophyllineTrimebutine MaleateVancomycin Hydrochloride

24. The following monographs were revised in ori-gin:Part I

AcetylspiramycinAclarubicin HydrochlorideActinomycin DAmoxicillinAmpicillinAnhydrous AmpicillinAmpicillin SodiumArbekacin SulfateAstromicin SulfateBacampicillin HydrochlorideBekanamycin SulfateBenzbromaroneBenzylpenicillin PotassiumBleomycin HydrochlorideBleomycin SulfateCalcium Chloride InjectionCarumonam SodiumCefaclorCefaloridinCefalotin SodiumCefamandole SodiumCefbuperazone SodiumCefmenoxime HydrochlorideCefotaxime SodiumCefotetanCefotiam Hexetil HydrochlorideCefoxitin SodiumCefpiramide SodiumCefroxadineCefteram PivoxilCefuroxime AxetilChloramphenicolCiclacillinCitric AcidAnhydrous Citric AcidClindamycin PhosphateDibekacin SulfateDoxorubicin HydrochlorideDoxycycline HydrochlorideErythromycinFlomoxef SodiumFradiomycin Sulfate

Gentamicin SulfateGlycerinConcentrated GlycerinGriseofulvinImipenemJosamycinJosamycin PropionateKanamycin SulfateLatamoxef SodiumLincomycin HydrochlorideMicronomicin SulfateMitomycin COxytetracycline HydrochloridePeplomycin SulfatePipemidic Acid TrihydratePivmecillinam HydrochloridePolymixin B SulfatePotassium ClavulanateProcarbazine HydrochloridePyrazinamideRetinol AcetateRetinol PalmitateRibostamycin SulfateRifampicinSodium ChlorideStreptomycin SulfateSulbenicillin SodiumTalampicillin HydrochlorideTeicoplaninTobramycinTrichomycinTrimetoquinol HydrochlorideVancomycin Hydrochloride

Part IIAloeCod Liver OilEvodia FruitGardenia FruitPueraria RootSenna LeafPowdered Senna LeafVitamin A OilVitamin A Oil Capsules

xx Supplement I, JP XIVPreface

Those who were engaged in the preparation of theSupplement I to JP Fourteenth Edition are as follows:

Norio AimiMitsuo AokiKiichi AonukiNobuo AoyagiYoshichika ArakawaKeiko ArimotoKazuhide AshizawaShinichiro AsoKunihiro FujitaHiroshi FujiwaraGoro FunamotoYukihiro GodaMorio HamashimaRuri HanajiriKouji HasegawaRyuichi HasegawaTakao HayakawaMasahiro HayashiFusayoshi HirayamaYukio HiyamaKunimoto HottaTakanori IchikawaNobukazu IgoshiToshio ImanariKenichi InuiMumio IshibashiShigeru ItaiMitsuo ItoYuji ItoTakashi ItohShozo IwagamiAkemi KaiKazuaki KakehiShozo KamiyaTakemine KanaiNahoko KaniwaMotoko Kanke

Yoshiaki KatoMitsunori KatohNoriko KatoriNobuo KawaharaToru KawanishiToshiaki KawanishiNana KawasakiToshisuke KawasakiYoshiaki KawashimaKeiji KijimaTakao KiyoharaToyohiko KobayashiMasayoshi KohaseShigeo KojimaHiroyasu KokuboSeizo KondoHideki KumakuraTakao KunisadaMitsuo KurashigeTakeshi KurataMasaaki KuriharaHaruo KuriyamaFumiyo KusuMasako MaedaTamio MaitaniToshio MasaokaToshihiko MatsubaraYoshihisa MatsudaNorio MatsukiShigeru MatsukiHayashi MatsukuraKatsutoshi MiseHiroto MiyamotoNaoki MiyataMichinao MizugakiKaoru MorikawaOsamu Morita

Takashi MoritaToshimi MuraiShigeru MurakiEmi NakajimaHiroshi NakamuraTatsuya NakanoHiroyuki NakazawaMasaaki NaotsukaMasao NasuKoji NishijimaMotohiro NishijimaTatsumi NishiyamaTakashi NomotoHiroyasu OgataYoshiyuki OgawaMasaru OhnoYasuo OhnoYoshiro OhtaniMasakazu OotaniMinoru OkadaSatoshi OkadaTsuneo OkuboHaruhiro OkudaMasami OtsukaTadashi OuchiKazuhiko SagaraEiji SakaiHideki SasakiTsuguo SasakiMotoyoshi SatakeAkihiro SatoSetsuko SekitaYasuo ShimadaKesamitsu ShimizuKyoko ShimuraFumitoshi ShinchoOsamu ShirotaKouichi Shudo

Hisashi SonobeShoko SueyoshiHisakazu SunadaHideyo SuzukiSenji SuzukiYukio TabuchiYoshikazu TakahashiTadahiro TakedaYasushi Takeda**Hiroshi TokunagaToyoshige TanabeHaruo TanakaToshihiro TanakaKenichi TanamotoTsuyoshi TanimotoSusumu TerabayashiTadao Terao*Hiroshi TerashimaKunikazu TeshimaKiyoshi TomiokaMotowo TomitaTatsuru TomizawaNobuchika TsumagariMitsuru Uchiyama*Yoshimasa UeharaTakashi UnnoMorimasa YagisawaTakehiko YajimaTeruhide YamaguchiKeiichi YamamotoKeiji YamamotoKenichi YamazakiTakeshi YamazakiHikaru YodenChikako YomotaHitoo YoshidaKazumasa YoshikawaSumie Yoshioka

*Chairman, Committee on JP**Acting Chairman, Committee on JP

13591359

General NoticesChange the paragraph 6 to read:

6. The following abbreviations are used for themain units.

meter mcentimeter cmmillimeter mmmicrometer mmnanometer nmkilogram kggram gmilligram mgmicrogram mgnanogram ngpicogram pgCelsius degree 9Csquare centimeter cm2

liter Lmilliliter mLmicroliter mLmegahertz MHzper centimeter cm－1

newton Nkilopascal kPamole per liter mol/Lmillipascal second mPa・ssquare millimeter second mm2/slux lxmass per cent z

mass parts per million ppmmass parts per billion ppbvolume per cent volzvolume parts per million vol ppmmass per volume per cent w/vz

hydrogen ion concentration pHEndotoxin unit EU

Note: ``ppm'' used in the Nuclear MagneticResonance Spectroscopy (1H) indicates the chemicalshift, and ``w/vz'' is used in the formula or compo-sition of preparations.

Change the paragraph 8 to read:

8. Standard temperature, ordinary temperature,room temperature, and lukewarm are deˆned as 209C,15 – 259C, 1 – 309C, and 30 – 409C, respectively. Acold place, unless otherwise speciˆed, shall be a placehaving a temperature of 1 – 159C.

The temperatures of cold water, lukewarm water,warm water, and hot water are deˆned as not exceed-ing 109C, 30 – 409C, 60 – 709C, and about 1009C,respectively.

The term ``heated solvent'' or ``hot solvent'' meansa solvent heated almost to the boiling point of the sol-vent, and the term ``warmed solvent'' or ``warm sol-vent'' usually means a solvent heated to a temperaturebetween 609C and 709C. The term ``heat on or in awater bath'' indicates, unless otherwise speciˆed,heating with a boiling water bath or a steam bath atabout 1009C.

Cold extraction and warm extraction are usuallyperformed at temperatures of 15 – 259C and 35 – 459C, respectively.

13611361

General Rulesfor Preparations

11. Injections

Change the paragraph (9) to read:

(9) Unless otherwise speciˆed, Injections meet therequirements of the Sterility Test. In the case of drugsto be dissolved before use, carry out the test with thesolution obtained by dissolving the contents in the at-tached solvent.

13631363

General Tests, Processesand Apparatus

Change to read:

General Tests, Processes and Apparatus includes commonmethods for tests and other articles related to them. Unlessotherwise speciˆed, the procedures for absorbance determi-nation, absorbance ratio determination, acid-neutralizingcapacity determination of gastrointestinal medicines, alco-hol number determination, ammonium determination, ar-senic determination, atomic absorption spectrophotometry,test for bacterial endotoxins, boiling point determination,distilling range determination, chloride determination, con-ductivity measurement, congealing point determination, testfor content uniformity, determination of bulk and tappeddensities, digestion test, disintegration test, dissolution test,endpoint detection in titrimetry, ‰ame coloration, ‰uoro-metry, foreign insoluble matter test for injections, gas chro-matography, heavy metals determination, infrared spec-trophotometry, insoluble particulate matter test for injec-tions, insoluble particulate matter test for ophthalmic solu-tions, iron determination, liquid chromatography, loss ondrying determination, loss on ignition determination, massvariation test, melting point determination, methanol deter-mination, methoxyl assay, test for microbial limit, test formicrobial limit for crude drugs, microbiological potency de-termination for antibiotics, mineral oil determination, nitro-gen determination, nuclear magnetic resonance spec-troscopy, optical rotation determination, osmolarity deter-mination, oxygen ‰ask combustion method, paper chro-matography, particle size distribution test for preparations,pH determination, powder particle size determination, testfor pyrogen, qualitative test, test for readily carbonizablesubstances, refractive index determination, residual solventstest, residue on ignition determination, speciˆc gravity anddensity determination, speciˆc surface area determination,test for sterility, sulfate determination, test for glass contain-ers for injections, test for metal particles in ophthalmic oint-ments, test for plastic containers, test for rubber closure foraqueous infusions, test for total organic carbon, thermalanalysis, thin-layer chromatography, viscosity determina-tion, vitamin A assay, test for volatile contaminants inethanol, water determination, and X-ray powder diŠractionare performed as directed in the corresponding articles underthe General Tests, Processes and Apparatus. The tests formelting point of fats, congealing point of fatty acids, speciˆcgravity, acid value, saponiˆcation value, ester value,hydroxyl value, unsaponiˆable matter and iodine value offats and fatty oils are performed as directed in the corre-sponding items under the Fats and Fatty oils Test, and thetests for foreign matter and loss on drying, total ash, acid-in-

soluble ash, extract content, essential oil content of crudedrugs are performed as directed in the corresponding itemsunder the Crude Drugs Test.

23. Infrared SpectrophotometryChange the Instrument and adjustment to read:

Instrument and adjustmentSeveral models of dispersive infrared spectrophotometers

or Fourier-transform infrared spectrophotometers are avail-able.

The instruments, adjusted according to the instructionmanual of each individual instrument, should comply withthe following test for resolving power, transmittance repro-ducibility and wave number reproducibility. When the spec-trum of a polystyrene ˆlm about 0.04 mm thick is recorded,the depth of the trough from the maximum absorption atabout 2850 cm－1 to the minimum at about 2870 cm－1 shouldbe not less than 18z transmittance and that from the maxi-mum at about 1583 cm－1 to the minimum at about 1589cm－1 should be not less than 12z transmittance.

The wave number (cm－1) scale is usually calibrated by theuse of several characteristic absorption wave numbers(cm－1) of a polystyrene ˆlm shown below. The number inparentheses indicates the permissible range.

3060.0 (±1.5) 2849.5 (±1.5) 1942.9 (±1.5)1601.2 (±1.0) 1583.0 (±1.0) 1154.5 (±1.0)1028.3 (±1.0)

When the dispersive infrared spectrophotometer is used,the permissible range of the absorption wave unmbers at1601.2 cm－1 and at 1028.3 cm－1 should be both within±2.0cm－1.

As the repeatability of transmittance and wave number,the diŠerence of transmittance should be within 0.5z whenthe spectrum of a polystyrene ˆlm is measured twice at sever-al wave numbers from 3000 to 1000 cm－1, and the diŠerenceof wave number should be withun 5 cm－1 at about 3000cm－1 and within 1 cm－1 at about 1000 cm－1.

Change to read:

52. Residue on Ignition Test

The Residue on Ignition Test is a method to measure themass of the residual substance not volatilized when the sam-ple is ignited with sulfuric acid by the method described be-

13641364 Supplement I, JP XIVGeneral Tests, Processes and Apparatus

low. Generally, this test is intended for determining the con-tent of inorganic substances contained as impurities in an or-ganic substance.

The description, for example, ``not more than 0.10z (1g),'' in a monograph, indicates that the mass of the residue isnot more than 1.0 mg per 1 g of the substance in the test inwhich about 1 g of the substance is weighed accurately andignited by the procedure described below, and `àfterdrying'' indicates that the sample is tested after being driedunder the conditions speciêd in the test for Loss on drying.

ProcedurePreviously ignite a crucible of platinum, quartz or por-

celain at 600± 509C for 30 minutes, and weigh accuratelyafter cooling in a desiccator (silica gel).

Take the sample of the amount directed in the mono-graph, transfer into the ignited crucible, and weigh accurate-ly. When the quantity of the sample to be taken is indicatedin a volume, pipet exactly the amount directed in the mono-graph and transfer into the above crucible. When directed as`àfter evaporating,'' heat properly to evaporate the solu-tion.

Moisten the sample with a small amount of sulfuric acid,usually 1 mL, then heat slowly at a temperature as low aspracticable until the sample is completely carbonized, andcool. Moisten again with a small amount of sulfuric acid,heat gently until white fumes are no longer evolved, and ig-nite at 600± 509C until the residue is completely incinerat-ed. Proceed with care to not burn with a ‰ame. Cool thecrucible in a desiccator (silica gel), and reweigh accurately tocalculate the amount of the residue.

When the amount of the residue obtained above exceedsthe limit speciêd in the monograph, unless otherwise speci-êd, ignite repeatedly to constant mass.

Change to read:

66. Vitamin A Assay

The Vitamin A Assay is a method to determine vitamin Ain Retinol Acetate, Retinol Palmitate, Vitamin A Oil, CodLiver Oil and other preparations. Method 1 is for the assayof synthetic vitamin A esters, using the ultraviolet-visiblespectrophotometry (Method 1-1) or the liquid chro-matography (Method 1-2). Method 2 is for the assay of vita-min A of natural origin, containing many geometricalisomers, using the ultraviolet-visible spectrophotometry todetermine vitamin A as vitamin A alcohol obtained bysaponiˆcation in an alkaline solution and extraction.

One Vitamin A Unit (equal to 1 vitamin A I.U.) is equiva-lent to 0.300 mg of vitamin A (all-trans vitamin A alcohol).

ProcedureAll procedures should be carried out quickly and care

should be taken as far as possible to avoid exposure to light,air, oxidants, oxidizing catalysts (e.g. copper, iron), acidsand heat. If necessary, light-resistant vessels may be used.

Generally, for synthetic vitamin A esters apply Method1-1 or Method 1-2, but if the assay conditions required forMethod 1-1 are not suitable, apply Method 2.

Method 1-1Weigh accurately about 0.1 g of the sample, and dissolve

in 2-propanol for vitamin A assay to make exactly 50 mL.Dilute this solution with 2-propanol for vitamin A assay tomake a solution so that each mL contains 10 to 15 vitamin AUnits, and use this solution as the sample solution. Deter-mine the absorption spectrum of the sample solution be-tween 220 nm and 400 nm as directed under the Ultraviolet-visible Spectrophotometry to obtain the wavelength of themaximum absorption and the absorbances at 300 nm, 310nm, 320 nm, 326 nm, 330 nm, 340 nm and 350 nm. Whenthe maximum absorption lies between 325 nm and 328 nm,and the ratios, Ali/A326, of each absorbance, Ali, at 300 nm,310 nm, 320 nm, 330 nm, 340 nm and 350 nm to the absor-bance, A326, at 326 nm are within the range of±0.030 of thevalues in the Table, the potency of vitamin A in Units per gof the sample is calculated from the following equation.

Units of vitamin A in 1 g＝A326

W×

V100

× 1900

A326: Absorbance at 326 nmV: Total volume (mL) of the sample solutionW: Amount (g) of sample in V mL of the sample solution1900: Conversion factor from speciˆc absorbance of

retinol ester to IU (Unit/g)

This method is applied to drugs or preparations contain-ing vitamin A esters (retinol acetate or retinol palmitate) asthe main component. However, when the wavelength ofmaximum absorption does not lie between 325 nm and 328nm, or when the absorbance ratio Ali/A326 is not within therange of±0.030 of the values in the Table, apply Method 2.

Table Absorbance Ratio, Ali/A326, of retinol acetate andretinol palmitate

li (nm)Ali/A326

Retinol acetate Retinol palmitate

300310320330340350

0.5780.8150.9480.9720.7860.523

0.5900.8250.9500.9810.7950.527

Method 1-2Proceed with an appropriate amount of sample as directed

under the Liquid Chromatography.For the assay of retinol acetate and retinol palmitate use

Retinol Acetate Reference Standard and Retinol PalmitateReference Standard, respectively, and ˆx appropriately theoperating procedure, the operating conditions and the sys-tem suitability based on the characteristics of the substanceto be tested and the species and amount of coexisting sub-stances.

13651365Supplement I, JP XIV General Tests, Processes and Apparatus

Method 2Unless otherwise speciˆed, weigh accurately a sample con-

taining not less than 500 Units of vitamin A, and not morethan 1 g of fat, transfer to a ‰ask, and add 30 mL of alde-hyde-free ethanol and 1 mL of a solution of pyrogallol inethanol (95) (1 in 10). Then add 3 mL of a solution of potas-sium hydroxide (9 in 10), attach a re‰ux condenser, and heaton a water bath for 30 minutes to saponify. Cool quickly toordinary temperature, add 30 mL of water, transfer to aseparator A, wash the ‰ask with 10 mL of water and then 40mL of diethyl ether, transfer the washings to the separatorA, shake well, and allow to stand. Transfer the water layerso obtained to a separator B, wash the ‰ask with 30 mL ofdiethyl ether, add the washing to the separator B, and ex-tract by shaking. Transfer the water layer to a ‰ask, add thediethyl ether layer to the separator A, transfer the water lay-er in the ‰ask to the separator B, add 30 mL of diethyl ether,and extract by shaking. Transfer the diethyl ether layer soobtained to the separator A, add 10 mL of water, allow theseparator A to stand after gentle turning upside-down 2 or 3times, and remove the water layer. Wash the content of theseparator A with three 50-mL portions of water with increas-ingly vigorous shaking as the washing proceeds. Furtherwash with 50-mL portions of water until the washing no lon-ger shows a pink color with phenolphthalein TS, and allowto stand for 10 minutes. Remove remaining water as far aspossible, transfer the diethyl ether to an Erlenmeyer ‰ask,wash the separator with two 10-mL portions of diethyl ether,add the washings to the ‰ask, add 5 g of anhydrous sodiumsulfate to the ‰ask, mix by shaking, and transfer the diethylether to a round-bottomed ‰ask by decantation. Wash theremaining sodium sulfate in the ‰ask with two or more10-mL portions of diethyl ether, and transfer the washingsto the ‰ask. Evaporate the diethyl ether in a water bath at 459C while swirling the ‰ask, using an aspirator, to about 1mL, immediately add an exactly measured amount of 2-propanol for vitamin A assay to make a solution containing6 to 10 vitamin A Units per mL, and designate the solutionas the sample solution. Determine the absorbances, A310 at310 nm, A325 at 325 nm, and A334 at 334 nm, of the samplesolution as directed under the Ultraviolet-visible Spec-trophotometry.

Units of vitamin A in 1 g of the sample

＝A325

W×

V100

× f× 1830

f＝ 6.815－ 2.555×A310

A325－ 4.260×

A334

A325

A325: Absorbance at 325 nmV: Total volume (mL) of the sample solutionW: Amount (g) of sample in V mL of the sample solutionf: Correction factor1830: Conversion factor from speciˆc absorbance of

retinol alcohol to IU (Unit/g)

70. Reference Standards;Reagents, Test Solutions; StandardSolutions for Volumetric Analysis;

Standard Solutions; MatchingFluids for Color; Optical Filters forWavelength and Transmission Rate

Calibration; and MeasuringInstruments, Appliances

(1) Reference Standards

Delete the following Reference Standards:

Drostanolone Propionate, Kitasamycin

Add the following:

Aceglutamide, Acetylspiramycin II, Aclarubicin, Actino-mycin D, Arbekacin Sulfate, Astromicin Sulfate, Bacitracin,Bekanamycin Sulfate, Benzylpenicillin Potassium, Benzyl-penicillin Sodium, Bleomycin A2 Hydrochloride, Carumon-am Sodium, Cefaclor, Cefaloridin, Cefalotin Sodium, Cefa-mandole Lithium, Cefbuperazone, Cefmenoxime Hydro-chloride, Cefodizime Sodium, Cefotaxime, Cefotetan,Cefotiam Hexetil Hydrochloride, Cefoxitin, Cefpiramide,Cefpodoxime Proxetil, Cefroxadine, Cefteram PivoxilMesit-ylenesulfonate, Cefuroxime Axetil, Chloramphenicol, Chlo-ramphenicolPalmitate,ChloramphenicolSuccinate,Ciclacil-lin, Clindamycin Hydrochloride, Clindamycin Phosphate,Colistin Sulfate, Daunorubicin Hydrochloride, Demethyl-chlortetracyclineHydrochloride,DibekacinSulfate,Diethan-olamine Fusidate, Doxorubicin Hydrochloride, DoxycyclineHydrochloride, Enviomycin Sulfate, Epirubicin Hydrochlo-ride, Flomoxef Triethylammonium, Fradiomycin Sulfate,Gentamicin Sulfate, Gramicidin, Griseofulvin, Imipenem,Josamycin Propionate, Kanamycin Monosulfate, LatamoxefAmmonium, Lenampicillin Hydrochloride, Leucomycin A5,Lincomycin Hydrochloride, Lysozyme, Micronomicin Sul-fate, Mitomycin C, Oxytetracycline Hydrochloride, Pepl-omycin Sulfate, L-Phenethicillin Potassium, Pimaricin,Pirarubicin, Pivmecillinam Hydrochloride, Polymixin B Sul-fate, Puerarin, Pyrrolnitrin, Ranitidine Hydrochloride,Retinol Acetate, Retinol Palmitate, Ribostamycin Sulfate,Rifampicin, Sennoside A, Sennoside B, Siccanin, Spec-tinomycin Hydrochloride, Streptomycin Sulfate, Sulbenicil-lin Sodium, Talampicillin Hydrochloride, Tobramycin,Trichomycin, Vancomycin Hydrochloride

(2) Reagents, Test Solutions

Delete the following Solution:

0.1 mol/L Phosphate buŠer solution for ceftibuten, pH8.0


Change the following:

Arecoline hydrobromide for thin-layer chromatographyC8H13NO2.HBr White crystals. Freely soluble in water,soluble in methanol, and practically insoluble in diethylether.

Melting point: 169 – 1719CPurity Related substances—Dissolve 50 mg of arecoline

hydrobromide for thin-layer chromatography in exactly 10mL of methanol. Perform the test with 10 mL of this solutionas directed in the Identiˆcation under Areca: any spot otherthan the principal spot at the Rf value of about 0.4 does notappear.

N-Demethylroxithromycin C40H74N2O15 White pow-der.

Identiˆcation—Determine the infrared absorption spec-trum of a solution of the substance to be tested in chlo-roform (1 in 20) as directed in the solution method under theInfrared Spectrophotometry using a 0.1-mm cell made ofpotassium bromide: it exhibits absorption at the wave num-bers of about 3600 cm－1, 3520 cm－1, 3450 cm－1, 3340 cm－1,1730 cm－1 and 1627 cm－1.

Geniposide for thin-layer chromatography C17H24O10

White crystals or crystalline powder. Melting point: 159 –1639C.

Purity Related substances—Dissolve 1.0 mg of genipo-side for thin-layer chromatography in exactly 1 mL ofmethanol, and perform the test with 20 mL of this solution asdirected in the Identiˆcation (2) under Gardenia Fruit: anyspot other than the principal spot at the Rf value of about0.3 does not appear.

Imidazole TS Dissolve 8.25 g of imidazole in 65 mL ofwater, adjust the pH to 6.8 with 5 mol/L hydrochloric acidTS, and add water to make 100 mL.

Add the following:

2-Acetamidoglutarimide C7H10N2O3: 170.17Identiˆcation—Determine the infrared absorption spec-

trum of 2-acetamidoglutarimide as directed in the potassiumbromide disk method under the Infrared Spectrophotomet-ry: it exhibits absorption at the wave numbers of about 3350cm－1, 1707 cm－1, 1639 cm－1 and 1545 cm－1.

Purity Related substances—Dissolve 10 mg of 2-acetamidoglutarimide in 100 mL of the mobile phase, anduse this solution as the sample solution. Pipet 1 mL of thesample solution, add the mobile phase to make exactly 100mL, and use this solution as the standard solution. Proceedwith 20 mL each of the sample solution and the standard so-lution as directed in the Purity (3) under Aceglutamide Alu-minum: the total of the peak areas other than 2-acetamidoglutarimide from the sample solution is not morethan the peak area from the standard solution.

Content: not less than 98.0z. Assay—Weigh accuratelyabout 20 mg of 2-acetamidoglutarimide, and perform thetest as directed under the Nitrogen Determination.

Each mL of 0.01 mol/L sulfuric acid VS＝ 0.8509 mg of C7H10N2O3

Acetaminophen C8H9NO2 [Same as the namesakemonograph]

Acetate buŠer solution, pH 5.4 To 5.78 mL of aceticacid (100) add water to make 1000 mL (solution A). Dissolve8.2 g of anhydrous sodium acetate in water to make 1000mL (solution B). Mix 176 mL of the solution A and 824 mLof the solution B, and adjust, if necessary, the pH to 5.4with the solution A or the solution B.

0.02 mol/L Acetic acid-sodium acetate TS Dissolve 2.74g of sodium acetate trihydrate in a suitable amount of water,and add 2 mL of acetic acid (100) and water to make 1000mL.

Albi‰orin C23H28O11.xH2O Colorless powder havingno odor. Freely soluble in water and in methanol, and prac-tically insoluble in diethyl ether.

Purity—Dissolve 1 mg in diluted methanol (1 in 2) tomake 10 mL, and use this solution as the sample solution.Perform the test with 10 mL of the sample solution as direct-ed in the Assay under Peony Root; the total area of thepeaks other than albi‰orin and other than the solvent is notlarger than 1/10 of the total area of the peaks other than thesolvent peak.

0.05 mol/L Ammonium formate buŠer solution, pH 4.0Dissolve 3.5 g of ammonium formate in about 750 mL ofwater, adjust the pH to 4.0 with formic acid, and add waterto make 1000 mL.

Amoxicillin C16H19N3O5S.3H2O [Same as the name-sake monograph]

a-Apooxytetracycline C22H22N2O8 Yellow-brown togreen powder.

Melting point: 200 – 2059C

b-Apooxytetracycline C22H22N2O8 Yellow-brown tobrown powder.

Purity Related substances—Dissolve 8 mg of b-apoox-ytetracycline in 5 mL of 0.01 mol/L sodium hydroxide TS,add 0.01 mol/L hydrochloric acid TS to make 100 mL, anduse this solution as the sample solution. Proceed the testwith 20 mL of the sample solution as directed in the Purity(2) under Oxytetracycline Hydrochloride, determine eachpeak area by the automatic integration method, and calcu-late the amounts of them by the area percentage method: thetotal amount of the peaks other than β-apooxytetracyclineis not more than 10z.

L-Arabinose C5H10O5 [K 8054: 1991, L(＋)-Arabinose,Special class]

Aristolochic acid I for crude drugs purity testC17H11NO7 Yellow crystalline powder. Melting point:about 2809C (with decomposition).

Absorbance E 1z1cm (318 nm): 384 – 424 (1 mg, methanol,

100 mL).Purity Related substances—Dissolve 1.0 mg of aristo-


lochic acid I for crude drugs purity test in 100 mL of dilutedmethanol (3 in 4), and use this solution as the sample solu-tion. Pipet 1 mL of this solution, add diluted methanol (3 in4) to make exactly 100 mL, and use this solution as the stan-dard solution. Perform the test with exactly 10 mL each ofthe sample solution and the standard solution as directed un-der the Liquid Chromatography according to the followingconditions, and determine each peak area by the automaticintegration method: the total area of the peaks other thanaristolochic acid I obtained from the sample solution is notmore than the peak area of aristolochic acid I from the stan-dard solution.Operating conditions

Detector, column, column temperature, mobile phase,and ‰ow rate: Proceed as directed in the operating condi-tions in the Purity (3) under Asiasarum Root.

Time span of measurement: About 3 times as long as theretention time of aristolochic acid I after the solvent peak.System suitability

Proceed as directed in the system suitability in the Purity(3) under Asiasarum Root.

Barbaloin for component determination Use barbaloinfor thin-layer chromatography meeting the following addi-tional speciˆcations.

Absorbance E 1z1cm (360 nm): 260 – 290 [10 mg dried in a

desiccator (in vacuum, phosphorus (V) oxide) for not lessthan 24 hours, methanol, 500 mL].

Purity Related substances—Dissolve 10 mg of the sub-stance to be tested in 10 mL of methanol, and use this solu-tion as the sample solution. Pipet 1 mL of the sample solu-tion, add methanol to make exactly 100 mL, and use this so-lution as the standard solution (1). Perform the test withexactly 20 mL each of the sample solution and the standardsolution (1) as directed under the Liquid Chromatographyaccording to the following conditions, and measure eachpeak area of the both solutions by the automatic integrationmethod: the total area of the peaks other than barbaloinfrom the sample solution is not larger than the peak area ofbarbaloin from the standard solution (1).Operating conditions

Proceed the operating conditions in the Component deter-mination under Aloe except wavelength, detection sensitivityand time span of measurement.

Wavelength: 300 nmDetection sensitivity: Pipet 1 mL of the standard solution

(1), add methanol to make exactly 20 mL, and use this solu-tion as the standard solution (2). Adjust the detection sen-sitivity so that the peak area of barbaloin obtained from 20mL of the standard solution (2) can be measured by the auto-matic integration method and the peak height of barbaloinobtained from 20 mL of the standard solution (1) showsabout 20z of the full scale.

Time span of measurement: About 3 times as long as theretention time of barbaloin after the solvent peak.

Becanamycin sulfate [Same as the namesake mono-graph]

Benzalphthalide C15H10O2 Yellow crystalline powder.Melting point: 99 – 1029C.

4-Chlorobenzenediazonium TS Dissolve 0.5 g of 4-chlo-roaniline in 1.5 mL of hydrochloric acid, and add water tomake 100 mL. To 10 mL of this solution add 10 mL of sodi-um nitrite TS and 5 mL of acetone. Prepare before use.

Cinnamic acid C9H8O2 White crystalline powder, hav-ing a characteristic odor.

Melting point: 132 – 1359C

Citric acid-phosphate-acetonitrile TS Dissolve 2.1 g ofcitric acid monohydrate, 13.4 g of dipotassium hydrogenphosphate and 3.1 g of potassium dihydrogen phosphate in1000 mL of a mixture of water and acetonitrile (3:1).

N-Demethylerythromycin C36H65NO13 White to lightyellowish white powder.

Deuterated formic acid for nuclear magnetic resonancespectroscopy DCOOD Prepared for nuclear magneticresonance spectroscopy.

Deuterated pyridine for nuclear magnetic resonance spec-troscopy C5D5N Prepared for nuclear magneticresonance spectroscopy.

Dibekacin sulfate [Same as the namesake monograph]

1,3-Dihydroxynaphthalene C10H8O2 Purple-brown,crystals or powder. Freely soluble in water and in ethanol(95).

Melting point: about 1259C

N,N-Dimethylacetamide CH3CON(CH3)2 Clear andcolorless liquid.

Boiling point: 163 – 1659CSpeciˆc gravity: 0.938 – 0.945 (Method 3).Water: not more than 0.2z (0.1 g, Coulometric titra-

tion).Purity—Perform the test with 3 mL of N,N-

dimethylacetamide as directed under the Gas Chro-matography according to the following conditions, deter-mine each peak area by the automatic integration method,and calculate the amount of N,N-dimethylacetamide by thearea percentage method: not less than 98.0z.Operating conditions

Detector: A hydrogen ‰ame-ionization detector.Column: A fused silica column 0.25 mm in inside di-

ameter and 30 m in length, coated the inside surface 0.5 mmin thickness with polyethylene glycol 20 M for gas chro-matography.

Column temperature: The sample is injected at a constanttemperature of about 709C, keep this temperature for 1minute, then raise to 2009C in a rate of 109C per minute,and keep 2009C for 3 minutes.

Carrier gas: HeliumFlow rate (linear velocity): About 30 cm/sec.Time span of measurement: About 2 times as long as the

retention time of N,N-dimethylacetamide.System suitability


Test for required detection: To exactly 1.0 g of N,N-dimethylacetamide add acetone to make exactly 100 mL.Pipet 5 mL of this solution, and add acetone to make exactly50 mL. Conˆrm that the peak area of N,N-dimethylacetamide obtained from 3 mL of this solution is e-quivalent to 40 to 60z of the full-scale.

System repeatability: When the test is repeated with 3 mLof N,N-dimethylacetamide under the above operating condi-tions, the relative standard deviation of the peak area ofN,N-dimethylacetamide is not more than 2.0z.

4-Dimethylaminoantipyrine C13H17N3O Colorless orwhite crystals, or a white crystalline powder.

Purity—Proceed the test with 5 mL of a solution of 4-dimethylaminoantipyrine (1 in 2000) as directed in the Assayunder Cefpiramide Sodium, determine each peak area in arange of about 2 times as long as the retention time of 4-dimethylaminoantipyrine after the solvent peak by the auto-matic integration method, and calculate the total amount ofthe peaks other than 4-dimethylaminoantipyrine by the areapercentage method: not more than 1.0z.

9,10-Diphenylanthracene C26H18 Yellow crystallinepowder. Soluble in diethyl ether, and practically insoluble inwater.


1,4-Diphenylbenzene C18H14 White scaly crystals, hav-ing a slight aromatic odor. It is freely soluble in ethanol(99.5), and slightly soluble in water.

Identiˆcation—Determine the infrared absorption spec-trum of 1,4-diphenylbenzene as directed in the potassiumbromide disk method under the Infrared Spectrophotomet-ry: it exhibits absorption at the wave numbers of about 3050cm－1, 3020 cm－1, 1585 cm－1, 1565 cm－1, 1476 cm－1, 1450cm－1, 995 cm－1, 834 cm－1, 740 cm－1 and 680 cm－1.

Divinylbenzene-methacrylate co-polymer for liquid chro-matography Prepared for liquid chromatography.

6-Epidoxycycline hydrochloride C22H24N2O8.HClYellow to dark yellow, crystals or crystalline powder.

Purity Related substances—Dissolve 20 mg of 6-epiox-ycycline hydrochloride in 25 mL of 0.01 mol/L hydrochloricacid TS, and use this solution as the sample solution. Pro-ceed the test with 20 mL of the sample solution as directed inthe Purity (2) under Doxycycline Hydrochloride, determineeach peak area by the automatic integration method, andcalculate the amounts of them by the area percentagemethod: the total area of the peaks other than 6-epidoxycy-cline is not more than 10z.

4-Epioxytetracycline C22H24N2O9 Green-brown to bro-wn powder.

Purity Related substances—Dissolve 20 mg of 4-epiox-ytetracycline in 25 mL of 0.01 mol/L hydrochloric acid TS,and use this solution as the sample solution. Proceed the testwith 20 mL of the sample solution as directed in the Purity(2) under Oxytetracycline Hydrochloride, determine eachpeak area by the automatic integration method, and calcu-late the amounts of them by the area percentage method: the

total amount of the peaks other than 4-epioxytetracycline isnot more than 10z.

Erythromycin B C37H67NO12 White to light yellowishwhite powder.

Purity Related substances—Dissolve 10 mg of erythro-mycin B in 1 mL of methanol, add a mixture of phosphatebuŠer solution, pH 7.0 and methanol (15:1) to make 5 mL,and use this solution as the sample solution. Pipet 1 mL ofthe sample solution, add a mixture of phosphate buŠer solu-tion, pH 7.0 and methanol (15:1) to make exactly 20 mL,and use this solution as the standard solution. Proceed withexactly 100 mL each of the sample solution and the standardsolution as directed in the Purity (3) Related substances un-der Erythromycin, and determine each peak area from thesolutions by the automatic integration method: the total ofareas of the peaks other than erythromycin B from the sam-ple solution is not more than the peak area of erythromycinB from the standard solution.

Erythromycin C C36H65NO13 White to light yellowishwhite powder.

Purity Related substances—Dissolve 10 mg of erythro-mycin C in 1 mL of methanol, add a mixture of phosphatebuŠer solution, pH 7.0 and methanol (15:1) to make 5 mL,and use this solution as the sample solution. Pipet 1 mL ofthe sample solution, add a mixture of phosphate buŠer solu-tion, pH 7.0 and methanol (15:1) to make exactly 20 mL,and use this solution as the standard solution. Proceed withexactly 100 mL each of the sample solution and the standardsolution as directed in the Purity (3) Related substances un-der Erythromycin, and determine each peak area from thesolutions by the automatic integration method: the total ofareas of the peaks other than erythromycin C from the sam-ple solution is not more than the peak area of erythromycinC from the standard solution.

3-Ethoxy-4-hydroxybenzaldehyde C9H10O3 White topale yellowish white crystalline. Freely soluble in ethanol(95), and slightly soluble in water.

Melting point: 76 – 789CContent: not less than 98.0z. Assay—Weigh accurately

about 0.3 g of 3-ethoxy-4-hydroxybenzaldehyde, previouslydried in a desiccator (phosphorous (V) oxide) for 4 hours,dissolve in 50 mL of N,N-dimethylacetamide, and titratewith 0.1 mol/L sodium methoxide VS (indicator: thymolblue TS).

Each mL of 0.1 mol/L sodium methoxide VS＝ 16.62 mg of C9H10O3

2-Formylbenzoic acid CHOC6H4COOH White crys-tals. Melting point: 97 – 999C

Content: not less than 99.0z. Assay—Weigh accuratelyabout 0.3 g of 2-formylbenzoic acid, previously dried (invacuum, phosphorus (V) oxide, 3 hours), dissolve in 50 mLof freshly boiled and cooled water, and titrate with 0.1mol/L sodium hydroxide VS (indicator: 3 drops of phenolred TS).


Each mL of 0.1 mol/L sodium hydroxide VS＝ 15.01 mg of C8H6O3

Geniposide for component determination Use genipo-side for thin-layer chromatography meeting the followingadditional speciˆcations.

Absorbance E 1z1cm (240 nm): 249 – 269 [10 mg dried in a

desiccator (reduced pressure of not exceeding 0.67 kPa,phosphorus (V) oxide) for 24 hours, diluted methanol (1 in2), 500 mL].

Purity Related substances—Dissolve 5 mg of geniposidefor component determination in 50 mL of diluted methanol(1 in 2), and use this solution as the sample solution. Pipet 1mL of the sample solution, add diluted methanol (1 in 2) tomake exactly 100 mL, and use this solution as the standardsolution (1). Perform the test with exactly 10 mL each of thesample solution and the standard solution (1) as directed un-der the Liquid Chromatography according to the followingconditions, and measure each peak area of the both solu-tions by the automatic integration method: the total area ofthe peaks other than geniposide from the sample solution isnot larger than the peak area of geniposide from the stan-dard solution (1).Operating conditions

Proceed as directed in the Component determination un-der Gardenia Fruit except detection sensitivity and time spanof measurement.

Detection sensitivity: Pipet 1 mL of the standard solution(1), add diluted methanol (1 in 2) to make exactly 20 mL,and use this solution as the standard solution (2). Adjust thedetection sensitivity so that the peak area of geniposide ob-tained from 10 mL of the standard solution (2) can be meas-ured by the automatic integration method and the peakheight of geniposide obtained from 10 mL of the standardsolution (1) shows about 20z of the full scale.

Time span of measurement: About 3 times as long as theretention time of geniposide after the solvent peak.

Guaifenesin C10H14O4 [Same as the namesake mono-graph]

Hirsutine C22H28N2O3 White to light yellow, crystalsor crystalline powder. Melting point: about 1009C.

Optical rotation [a]20D : about ＋579(10 mg, methanol, 1

mL).Absorbance E 1z

1cm (245 nm): 354 – 379 (5 mg calculatedon the anhydrous basis, a mixture of methanol and diluteacetic acid (7:3), 500 mL).

Purity Related substances－Dissolve 5 mg of hirsutinein 100 mL of a mixture of methanol and dilute acetic acid(7:3), proceed with 20 mL of this solution as directed in theComponent determination under Uncaria Thorn, and per-form the Liquid Chromatography: the sum of the peak areasexcept the areas of hirsutine and the solvent is not more than1/10 of the sum of the peak areas except the solvent.

0.01 mol/L Hydrochloric acid-methanol TS To 20 mLof 0.5 mol/L hydrochloric acid TS add methanol to make1000 mL.

0.05 mol/L Hydrochloric acid-methanol TS To 100 mLof 0.5 mol/L hydrochloric acid add methanol to make 1000mL.

1-(2-Hydroxyethyl)-1H-tetrazol-5-thiol C3H6N4OSWhite, crystals or powder.

Melting point: 136 – 1419CPurity Related substances—Dissolve 0.10 g of 1-(2-

hydroxyethyl)-1H-tetrazol-5-thiol in 1 mL of water, and usethis solution as the sample solution. Pipet 0.5 mL of thesample solution, add water to make exactly 25 mL, and usethis solution as the standard solution. Perform the test withthese solutions as directed under the Thin-layer Chro-matography. Spot 1 mL each of the sample solution and thestandard solution on a plate of silica gel with ‰uorescent in-dicator for thin-layer chromatography, develop with a mix-ture of ethyl acetate, water, methanol and formic acid(60:10:7:6) to a distance of about 10 cm, and air-dry theplate. Examine under ultraviolet light (main wavelength: 254nm): the spot other than the principal spot obtained fromthe sample solution is not more intense than the spot fromthe standard solution.

Isoniazid C6H7N3O [Same as the namesake mono-graph]

Josamycin C42H69NO15 [Same as the namesake mono-graph]

Josamycin propionate C45H73NO16 [Same as thenamesake monograph]

Lactobionic acid C12H22O12 Colorless crystals or whitecrystalline powder, having no odor.

Melting point: 113 – 1189CPurity—Dissolve 0.10 g of lactobionic acid in 10 mL of a

mixture of methanol and water (3:2), and perform the testwith 10 mL of this solution as directed in the Identiˆcation(2) under Erythromycin Lactobionate: the spot other thanthe principal spot is not found.

Luteolin for thin-layer chromatography C15H10O6

Light yellow to yellow-brown crystalline powder. Slightlysoluble in methanol and in ethanol (99.5), and practically in-soluble in water. Melting point: about 3109C (with decom-position).

Purity Related substances—Dissolve 1.0 mg of luteolinfor thin-layer chromatography in 1 mL of methanol. Pro-ceed the test with 10 mL of this solution as directed in theIdentiˆcation under Chrysanthemum Flower: any spot otherthan the principal spot of Rf about 0.7 does not appear.

D-Mannose C6H12O6 White crystal or crystalline pow-der. It is very soluble in water.

Melting point: about 1329C (with decomposition).Optical rotation [a]20

D : ＋13.7 –＋14.79(4 g, diluted am-monia TS (1 in 200), 20 mL, 100 mm).

Metacycline hydrochloride C22H22N2O8.HCl Yellowto dark yellow, crystals or crystalline powder.

Purity Related substances—Dissolve 20 mg of metacy-cline hydrochloride in 25 mL of 0.01 mol/L hydrochloric


acid TS, and use this solution as the sample solution. Pro-ceed the test with 20 mL of the sample solution as directed inthe Purity (2) under Doxycycline Hydrochloride, determineeach peak area by the automatic integration method, andcalculate the amounts of them by the area percentagemethod: the total area of peaks other than metacycline is notmore than 10z.

1-Methyl-1H-tetrazole-5-thiol for liquid chromatographyC2H4N4S White, crystals or crystalline powder. Very solu-ble in methanol, and freely soluble in water.

Melting point: 123 – 1279CLoss on drying: not more than 1.0z (1 g, in vacuum,

phosphorous (V) oxide, 2 hours).Content: not less than 99.0z. Assay—Weigh accurately

about 0.2 g of 1-methyl-1H-tetrazole-5-thiol, previously d-ried, dissolve in 80 mL of N,N-dimethylformamide, and ti-trate with 0.1 mol/L sodium methoxide VS (indicator: 3drops of thymol blue-N,N-dimethylformamide TS). Per-form a blank determination, and make any necessary correc-tion.

Each mL of 0.1 mol/L sodium methoxide VS＝ 11.61 mg of C2H4N4S

Morphine Hydrochloride [Same as the namesake mono-graph]

2-Naphthalenesulfonic acid C10H8O3S.H2O White topale yellowish white powder. Very soluble in water, inmethanol and in ethanol (95), and sparingly soluble indiethyl ether and in chloroform.

Water: 7.0 – 11.5z (0.5 g, volumetric titration, directtitration).

Content: not less than 95.0z, calculated on the anhy-drous basis. Assay—Weigh accurately about 0.5 g of 2-naphthalenesulfonic acid, dissolve in 30 mL of water, and ti-trate with 0.1 mol/L sodium hydroxide VS (indicator: 3drops of bromothymol blue TS). Perform a blank determi-nation, and make any necessary correction.

Each mL of 0.1 mol/L soduim hydroxide VS＝ 20.82 mg of C10H8O3S

1-Naphthol-sulfuric acid TS Dissolve 1.5 g of 1-naphthol in 50 mL of ethanol (95), add 3 mL of water and 7mL of sulfuric acid, and mix well. Prepare before use.

Ninhydrin-acetic acid TS Dissolve 1.0 g of ninhydrin in50 mL of ethanol (95), and add 10 mL of acetic acid (100).

Octadecylsilanized silica gel for thin-layer chro-matography Octadecylsilanized silica gel prepared forthin-layer chromatography.

Octadecylsilanized silica gel with ‰uorescent indicator forthin-layer chromatography Octadecylsilanized silica gelfor thin-layer chromatography containing ‰uorescent indi-cator.

O‰oxacin demethyl substance (±)-9-Fluoro-2,3-dihydro-3-methyl-7-oxo-7H-10-(1-piperazinyl)-pirido[1, 2, 3-de] [1,4]benzoxazine-6-carboxylic acid C17H18FN3O4 White

to light green-yellowish white, crystals or crystalline powder.Identiˆcation—Determine the infrared absorption spec-

trum of o‰oxacin demethyl substance as directed in thepotassium bromide disk method under the Infrared Spec-trophotometry: it exhibits absorption at the wave numbersof about 3050 cm－1, 2840 cm－1, 1619 cm－1, 1581 cm－1,1466 cm－1, 1267 cm－1, 1090 cm－1, 1051 cm－1 and 816cm－1.

Phenol red TS, dilute To 235 mL of a solution of ammo-nium nitrate (1 in 9400) add 105 mL of 2 mol/L sodiumhydroxide TS and 135 mL of a solution prepared by dissolv-ing 24 g of acetic acid (100) in water to make 200 mL. To thissolution add 25 mL of a solution prepared by dissolving 33mg of phenol red in 1.5 mL of 2 mol/L sodium hydroxideTS and adding water to make 100 mL. If necessary, adjustthe pH to 4.7.

50z Phenyl-50z methylpolysiloxane for gas chro-matography Prepared for gas chromatography.

0.01 mol/L Phosphate buŠer solution, pH 6.8 Dissolve1.36 g of potassium dihydrogen phosphate in 900 mL ofwater, adjust the pH to 6.8 with 0.2 mol/L sodiumhydroxide TS, and add water to make 1000 mL.

0.03 mol/L Phosphate buŠer solution, pH 7.5 Dissolve4.083 g of potassium dihydrogen phosphate in 800 mL ofwater, adjust the pH to 7.5 with 0.2 mol/L sodiumhydroxide TS, and add water to make 1000 mL.

1/15 mol/L Phosphate buŠer solution, pH 5.6 Dissolve9.07 g of potassium dihydrogen phosphate in about 750 mLof water, adjust the pH to 5.6 with potassium hydroxide TS,and add water to make 1000 mL.

0.1 mol/L Phosphate buŠer solution, pH 5.3 Dissolve0.44 g of disodium hydrogen phosphate 12-water and 13.32g of potassium dihydrogen phosphate in 750 mL of water,adjust the pH to 5.3 with sodium hydroxide TS or phosphor-ic acid, and add water to make 1000 mL.

0.1 mol/L Phosphate buŠer solution, pH 6.8 Dissolve6.4 g of potassium dihydrogen phosphate and 18.9 g of diso-dium hydrogen phosphate 12-water in about 750 mL ofwater, adjust the pH to 6.8 with sodium hydroxide TS ifnecessary, and add water to make 1000 mL.

0.1 mol/L Phosphate buŠer solution for antibiotics, pH8.0 Dissolve 16.73 g of dipotassium hydrogen phosphateand 0.523 g of potassium dihydrogen phosphate in about750 mL of water, adjust the pH to 8.0 with phosphoric acid,and add water to make 1000 mL.

Phosphate buŠer solution, pH 3.1 Dissolve 136.1 g ofpotassium dihydrogen phosphate in 500 mL of water, andadd 6.3 mL of phosphoric acid and water to make 1000 mL.

Phosphate buŠer solution, pH 5.9 Dissolve 6.8 g ofpotassium dihydrogen phosphate in 800 mL of water, adjustthe pH to 5.9 with diluted potassium hydroxide TS (1 in 10),and add water to make 1000 mL.


Phosphate buŠer solution, pH 6.2 Dissolve 9.08 g ofpotassium dihydrogen phosphate in 1000 mL of water (solu-tion A). Dissolve 9.46 g of disodium hydrogen phosphate in1000 mL of water (solution B). Mix 800 mL of the solutionA and 200 mL of the solution B, and adjust the pH to 6.2with the solution A or the solution B if necessary.

Phosphate buŠer solution for antibiotics, pH 6.5Dissolve 10.5 g of disodium hydrogen phosphate 12-waterand 5.8 g of potassium dihydrogen phosphate in 750 mL ofwater, adjust the pH to 6.5 with sodium hydroxide TS, andadd water to make 1000 mL.

Phosphate TS Dissolve 2.0 g of dipotassium hydrogenphosphate and 8.0 g of potassium dihydrogen phosphate inwater to make 1000 mL.

Phthalein purple C32H32N2O12.xH2O Yellowish whiteto brown power. Soluble in ethanol (95), and practically in-soluble in water.

Sensitivity test—Dissolve 10 mg of phthalein purple in 1mL of ammonia solution (28), and add water to make 100mL. To 5 mL of this solution add 95 mL of water, 4 mL ofammonia solution (28), 50 mL of ethanol (95) and 0.1 mL ofdiluted barium chloride TS (1 in 5): the solution shows ablue-purple color which disappears on the addition of 0.15mL of 0.1 mol/L disodium dihydrogen ethylenediaminetetraacetate TS.

Phthalimide C8H5NO2 White to pale brown crystals orpowder.

Melting point: 232 – 2379CClarity—1.0 g of phthalimide dissolves in 20 mL of sodi-

um hydroxide TS as a slight turbid solution.Content: not less than 98.0z. Assay—Weigh accurately

about 0.3 g of the substance to be tested, dissolve in 40 mLof N,N-dimethylformamide, and titrate with 0.1 mol/L so-dium methoxide VS (potentiometric titration). Perform ablank determination, and make any necessary correction.

Each mL of 0.1 mol/L sodium methoxide VS＝ 14.71 mg of C8H5NO2

Polyethylene glycol 600 for gas chromatography Pre-pared for gas chromatography.

Polyethylene glycol 1500 for gas chromatography Pre-pared for gas chromatography.

Polyoxyethylene (23) lauryl etherC12H25(OCH2CH2)nOH White masses. Melting point:about 409C

Potassium chloride for conductivity measurement[K 8121, Potassium chloride for conductivity measurement]

0.1 mol/L Potassium dihydrogen phosphate TS, pH 2.0Dissolve 13.6 g of potassium dihydrogen phosphate in waterto make 1000 mL. Adjust the pH to 2.0 with phosphoricacid.

0.25 mol/L Potassium dihydrogen phosphate TS, pH 3.5Dissolve 34 g of potassium dihydrogen phosphate in 900 mL

of water, adjust the pH to 3.5 with phosphoric acid, and addwater to make 1000 mL.

0.33 mol/L Potassium dihydrogen phosphate TS Dis-solve 4.491 g of potassium dihydrogen phosphate in water tomake 100 mL.

Potassium iodide TS, saturated Saturate 20 g of potassi-um iodide in 10 mL of ‰eshly boiled and cooled water. Pre-pare before use.

2-Propanol for vitamin A assay (CH3)2CHOH [K8839, Special class] When the absorbances at 300 nm andbetween 320 nm and 350 nm are determined as directed un-der the Ultraviolet-visible Spectrophotometry, using wateras the control, they are not more than 0.05 and not morethan 0.01, respectively. If necessary, purify by distillation.

Ranitidinediamine (C10H18N2OS)2.C4H4O4 White topale yellow crystalline powder.

Identiˆcation—Determine the infrared absorption spec-trum of ranitidinediamine as directed in the paste methodunder the Infrared Spectrophotometry: it exhibits absorp-tion at the wave numbers of about 2780 cm－1, 1637 cm－1,1015 cm－1 and 788 cm－1.

Content: not less than 95z. Assay—Weigh accuratelyabout 0.1 g of ranitidinediamine, dissolve in 50 mL of aceticacid (100), and titrate with 0.1 mol/L perchloric acid VS un-til the color of the solution changes from purple to greenthrough blue (indicator: crystal violet TS). Perform theblank determination in the same manner, and make anynecessary correction.

Each mL of 0.1 mol/L perchloric acid VS＝ 13.62 mg of (C10H18N2OS)2.C4H4O4

Resibufogenin for thin-layer chromatographyC24H32O4.nH2O White crystalline powder having no odor.It is freely soluble in acetone and in methanol.

Purity Related substances—Dissolve 5.0 mg of the sub-stance to be tested in exactly 5 mL of acetone. Perform thetest with 5 mL of this solution as directed in the Identiˆcationunder Toad Venom: no other spots than the principal spotof around Rf 0.4 appear.

L-Rhamnose monohydrate C6H12O.H2O White crys-talline powder having sweet taste. Freely soluble in water,and sparingly soluble in ethanol (95).

Optical rotation [a]20D :＋7.8 –＋8.39 (1 g, 20 mL of

water, 2 drops of ammonia TS, 100 mm).Melting point: 87 – 919CPurity Related substances—Dissolve 1.0 mg of L-rham-

nose monohydrate in 1 mL of water, and add methanol tomake exactly 10 mL. Proceed with 20 mL of this solution asdirected in the Identiˆcation (2) under Acacia: any spotother than the principal spot at the Rf value of about 0.5does not appear.

Rhynchophylline for component determinationC22H28N2O4 White, crystals or crystalline powder. Spar-ingly soluble in ethanol (99.5) and in acetone, and practicallyinsoluble in water. Melting point: 205 – 2099C


Absorbance E 1z1cm (245 nm): 473 – 502 (5 mg dried in a

desiccator (silica gel) for 24 hours, a mixture of methanoland dilute acetic acid (7:3), 500 mL).

Purity Related substances—(1) Dissolve 1.0 mg of rhynchophylline for component

determination in 1 mL of acetone, and perform the test withthis solution as directed under the Thin-layer Chro-matography. Spot 10 mL of the solution on a plate of silicagel with ‰uorescent indicator for thin-layer chro-matography, develop with a mixture of 1-butanol, water andacetic acid (100) (7:2:1) to a distance of about 10 cm, andair-dry the plate. Examine under ultraviolet light (mainwavelength: 254 nm): any spot other than the principal spotof Rf about 0.5 does not appear.

(2) Dissolve 5 mg of rhynchophylline for component de-termination in 100 mL of a mixture of methanol and diluteacetic acid (7:3), and use this solution as the sample solution.Pipet 1 mL of the sample solution, add a mixture ofmethanol and dilute acetic acid (7:3) to make exactly 100mL, and use this solution as the standard solution. Performthe test with exactly 20 mL each of the sample solution andthe standard solution as directed under the Liquid Chro-matography according to the following conditions. Deter-mine each peak area obtained from these solutions by theautomatic integration method: the sum of the peak areas ex-cept the areas of rhynchophylline and the solvent obtainedfrom the sample solution is not more than the peak area ofrhynchophylline from the standard solution.Operating conditions

Detector, column, column temperature, mobile phase,and ‰ow rate: Proceed as directed in the operating condi-tions in the Component determination under UncariaThorn.

Time span of measurement: About 4 times as long as theretention time of rhynchophylline after the solvent peak.System suitability

Test for required detectability: Measure exactly 1 mL ofthe standard solution, add a mixture of methanol and diluteacetic acid (7:3) to make exactly 20 mL. Conˆrm that thepeak area of rhynchophylline obtained from 20 mL of thissolution is equivalent to 3.5 to 6.5z of that from 20 mL ofthe standard solution.

System performance, and system repeatability: Proceed asdirected in the operating conditions in the Component deter-mination under Uncaria Thorn.

Sodium dihydrogen phosphate TS, pH 2.5 Dissolve 2.7g of sodium dihydrogen phosphate dihydrate in 1000 mL ofwater, and adjust the pH to 2.5 with phosphoric acid.

6 mol/L Sodium hydroxide TS Dissolve 252 g of sodiumhydroxide in water to make 1000 mL. Preserve in a polyethy-lene bottle.

Sodium 1-methyl-1H-tetrazole-5-thiolateC2H3N4NaS.2H2O White, crystals or crystalline powder.

Melting point: 90 – 949CPurity Related substances—Dissolve 10 mg of sodium 1-

methyl-1H-tetrazole-5-thiolate in 10 mL of water, and use

this solution as the sample solution. Perform the test withthis solution as directed under the Thin-layer Chro-matography. Spot 5 mL of the sample solution on a plate ofsilica gel with ‰uorescent indicator for thin-layer chro-matography. Develop the plate with a mixture of ethylacetate, acetone, water and acetic acid (100) (10:2:1:1) to adistance of about 10 cm, and air-dry the plate. Examine un-der ultraviolet light (main wavelength: 254 nm): any spotother than the principal spot does not appear.

Sodium 2-naphthalenesulfonate C10H7NaO3S Palebrown, crystals or powder.

Content: not less than 98.0z.

Sodium periodate TS Dissolve 60.0 g of sodium perio-date in 120 mL of 0.05 mol/L sulfuric acid TS, and addwater to make 1000 mL. If the solution is not clear, ˆlter thisthrough a glass-ˆlter. Keep in a light-resistant vessel.

Strongly acidic ion-exchange silica gel for liquid chro-matography Prepared for liquid chromatography.

Substrate solution for lysozyme hydrochloride To a suit-able amount of dried cells of Micrococcus luteus add a suita-ble amount of phosphate buŠer solution, pH 6.2, gentlyshake to make a suspension, and add the substrate cells orthe same buŠer solution so that the absorbance of the sus-pension at 640 nm is about 0.65. Prepare before use.

Tetracycline Hydrochloride C22H24N2O8.HCl Yellow,crystals or crystalline powder.

Purity Related substances—Dissolve 20 mg of tetracy-cline hydrochloride in 25 mL of 0.01 mol/L hydrochloricacid TS, and use this solution as the sample solution. Pro-ceed the test with 20 mL of the sample solution as directed inthe Purity (2) under Oxytetracycline Hydrochloride, deter-mine each peak area by the automatic integration method,and calculate the amounts of them by the area percentagemethod: the total amount of the peaks other than tetracy-cline is not more than 10z.

Tetra‰uoroethylene polymer for gas chromatographyPrepared for gas chromatography.

Tetrahydrofuran for liquid chromatography C4H8OClear and colorless liquid.

Refractive index n20D : 1.406 – 1.409

Density (209C): 0.884 – 0.889 g/mLPurity Ultraviolet absorbing substances—Determine the

absorption spectrum of tetrahydrofuran for liquid chro-matography as directed under the Ultraviolet-visible Spec-trophotometry, using water as the blank: the absorbences at240 nm, 254 nm, 280 nm, 290 nm, and between 300 nm and400 nm are not more than 0.35, 0.20, 0.05, 0.02 and 0.01, re-spectively.

Peroxide—Perform the test according to the method de-scribed in JIS K 9705: not more than 0.01z.

Tetrakishydroxypropylethylenediamine for gas chro-matography Prepared for gas chromatography.


Tetra-n-propylammonium bromide[CH3CH2CH2]4NBr White, crystals or crystalline powder.

Purity Clarity and color of solution—Dissolve 1.0 g oftetra-n-propylammonium bromide in 20 mL of water: thesolution is clear and colorless.

Content: not less than 98.0z. Assay—Weigh accuratelyabout 0.4 g of tetra-n-propylammonium bromide, dissolvein 50 mL of water, add 5 mL of dilute nitric acid, and titratewith 0.1 mol/L silver nitrate VS while shaking strongly(potentiometric titration).

Each mL of 0.1 mol/L silver nitrate VS＝ 26.63 mg of C12H28NBr

Thymine C5H6N2O2: 126.11Identiˆcation—Determine the infrared absorption spec-

trum of thymine, previously dried at 1059C for 3 hours, asdirected in the potassium bromide disk method under the In-frared Spectrophotometry: it exhibits absorption at the wavenumbers of about 3030 cm－1, 1734 cm－1, 1676 cm－1, 1446cm－1 and 814 cm－1.

Purity Related substances—Dissolve 50 mg of thyminein 100 mL of methanol. To 10 mL of this solution add themobile phase to make 100 mL, and use this solution as thesample solution. Proceed with 10 mL of the sample solutionas directed in the Purity (3) under Aceglutamide Aluminum:any peak does not appear at the retention time ofaceglutamide.

Tin (II) chloride-hydrochloric acid TS To 20 g of tin add85 mL of hydrochloric acid, heat until hydrogen gas no lon-ger are evolved, and allow to cool. Mix 1 volume of this so-lution and 10 volume of dilute hydrochloric acid. Preparebefore use.

Triethylamine buŠer solution, pH 3.2 To 4 mL oftriethylamine add 2000 mL of water, and adjust the pH to3.2 with phosphoric acid.

Trimethylsilanized silica gel for liquid chromatographyPrepared for liquid chromatography.

Water, sterile puriêd [Same as the namesake mono-graph in Part II]

(3) Standard Solutionsfor Volumetric Analysis

Add the following:

Barium chloride, 0.1 mol/L1000 mL of this solution contains 24.426 g of barium

chloride dihydrate (BaCl2.2H2O: 244.26).Preparation—Dissolve 24.5 g of barium chloride dihy-

drate in water to make 1000 mL, and standardize the solu-tion as follows:

Standardization—Measure exactly 20 mL of the preparedsolution, add 3 mL of hydrochloric acid, and warm the mix-ture. Add 40 mL of diluted sulfuric acid (1 in 130), previous-ly warmed, heat the mixture on a water bath for 30 minutes,and allow it to stand overnight. Filter the mixture, wash the

precipitate on the ˆlter paper with water until the last wash-ing shows no turbidity with silver nitrate TS, transfer theprecipitate together with the ˆlter paper to a tared crucible,and then heat strongly to ashes. After cooling, add 2 dropsof sulfuric acid, and heat again at about 7009C for 2 hours.After cooling, weigh accurately the mass of the residue, andcalculate the molarity factor as barium sulfate (BaSO4).

Each mL of 0.1 mol/L barium chloride VS＝ 23.34 mg of BaSO4

Add the following:

72. Conductivity Measurement

Conductivity measurement is a method for the measuringthe ‰owability of electric current in an aqueous solution.The measurement is made with a conductivity meter or a re-sistivity meter, and is used, for example, in the purity tests inmonographs. The method is applied to evaluate the test item``Conductivity (Electrical Conductivity)'' speciêd in themonographs. Further it is also used for monitoring the qual-ity of water in the preparation of highly puriêd water.However, when applying this method for monitoring thequality of water, the details of measurement should be speci-êd by the user, based on the principles described here.

Conductivity of a solution k (S・m－1) is deˆned as thereciprocal of resistivity r (Q・m), which is an indicator of thestrength of ionic conductivity for a ‰uid conductor. Resistiv-ity is deˆned as the product of electrical resistance per unitlength and cross-sectional area of a conductor. When resis-tivity is r, cross-section area A (m2), and length l (m),resistance R (Q) can be expressed by the following equation.

R＝ r(l/A )

Thus, conductivity k is expressed as follows,

k＝ 1/r＝ (1/R )(l/A )

If l/A is known, the conductivity k can be obtained bymeasuring resistance R or conductance G (＝ R－1).

In the International System (SI), the unit of conductivityis the Siemens per meter (S・m－1). In practice, conductivityof a solution is generally expressed by mS・cm－1, and resistiv-ity by Q・cm.

Unless otherwise speciêd, the reference temperature forthe expression of conductivity or resistivity is 209C.

ApparatusA conductivity meter or a resistivity meter is composed of

an indicator part (operating panel, display, recording unit)and a detector part, the latter of which includes a conductivi-ty cell. In the conductivity cell a pair of platinum electrodesis embedded. The cell is immersed in a solution, and theresistance or the resistivity of the liquid column between theelectrodes is measured. Alternating current is supplied tothis apparatus to avoid the eŠects of electrode polarization.Further, a temperature compensation system is generally


contained in the apparatus.Conductivity measurement is generally performed by us-

ing an immersion-type cell. A pair of platinum electrodes,the surfaces of which are coated with platinum black, is ˆxedin parallel. Both electrodes are generally protected by a glasstube to prevent physical shocks.

When the surface area of the electrode is A (cm2), and theseparation distance of the two electrodes is l (cm), the cellconstant C (cm－1) is given by the following equation.

C＝ a・(l/A )

a is a dimensionless numerical coe‹cient, and it is character-istic of the cell design.

In addition to the immersion-type cell, there are ‰ow-through-type and insert-in-pipe-type cells. These cells are setor inserted in an appropriate position in the ‰ow system formonitoring the quality of water continuously or intermit-tently, during the preparation of highly puriêd water.

Standard Solution of Potassium ChlorideAfter pulverizing an appropriate amount of potassium

chloride for conductivity measurement, dry it at 500 – 6009Cfor 4 hours. Take an indicated amount of the dried potassi-um chloride, as shown in Table 1, dissolve it in distilled orpuriêd water (conductivity less than 2 mS・cm－1), previous-ly boiled and cooled, and adjust to make 1000.0 g, forpreparation of the standard solutions. The conductivity andthe resistivity of the respective standard solutions at 209Care indicated in Table 1. These standard solutions should bekept in tightly closed polyethylene or hard glass bottles.

Table 1. Conductivity and Resistivity of the StandardSolutions of Potassium Chloride at 209C

Concentration Conductivity k Resistivity r(g/1000.0 g) (mS・cm－1) (Q・cm)

0.7455 1330 7520.0746 133.0 75190.0149 26.6 37594

When measurement at 209C can not be done, the indicat-ed value of conductivity for the respective standard solution(Table 1), can be corrected by using the equation below.However, the equation is valid only within the range of 20±59C.

kT＝ k20[1＋ 0.021(T－ 20)]

T: Measuring temperature speciêd in the monographkT: Calculated conductivity of the KCl standard solution

at T9Ck20: Conductivity of the KCl standard solution at 209C

Operating Procedure(1) Cell ConstantAn appropriate conductivity cell should be chosen accord-

ing to the expected conductivity of the sample solution. Thehigher the expected conductivity, the larger the cell constantrequired for the conductivity cell, so that the electricalresistance is within the measuring range of the apparatusbeing used. Conductivity cells with a cell constant of the ord-

er of 0.1 cm－1, 1 cm－1, or 10 cm－1, are generally used.For determination or conˆrmation of the cell constant, an

appropriate KCl standard solution should be chosen andprepared, taking account of the expected conductivity of thesample solution to be measured. Rinse the cell several timeswith distilled water. Next, after rinsing the cell 2 – 3 timeswith the standard solution used for the cell constant determi-nation, immerse the cell in the standard solution containedin a measuring vessel. After conˆrming that the temperatureof the standard solution is maintained at 20± 0.19C or atthe temperature speciêd in the monograph, measure theresistance RKCl or the conductance GKCl of the standard solu-tion, and calculate the cell constant C (cm－1) by use of thefollowing equation.

C＝ RKCl・kKCl or C＝ kKCl/GKCl

RKCl: Measured resistance (MQ)GKCl: Measured conductance (mS)kKCl: Conductivity of the standard solution being used

(mS・cm－1)

The measured cell constant should be consistent with thegiven value within 5z. If it is not consistent, coat the elec-trodes with platinum black again, or replace the cell with anew one.

(2) Suitability Test for the ApparatusUsing an appropriate KCl standard solution according to

the expected conductivity of the sample solution, performthe suitability test for the apparatus. Rinse the conductivitycell several times with distilled water, and rinse again 2 – 3times with the selected standard solution. Fill the standardsolution in the measuring vessel. After conˆrming that thetemperature of the measuring system is maintained at 20±0.19C, measure the conductivity of the standard solution.When this measuring procedure is repeated several times, theaverage conductivity should be consistent with an indicatedvalue in Table 1 within 5z. Further, the relative standarddeviation should be less than 2z.

(3) MeasurementAfter conˆrmation of the suitability of the apparatus, per-

form the conductivity measurement for the sample solution.Unless otherwise speciêd, the preparation method for sam-ple solution should be as speciêd in the respective mono-graph. Rinse the conductivity cell several times with distilledwater, and rinse again 2 – 3 times with sample solution. Im-merse the cell in the sample solution placed in a measuringvessel. If necessary, agitate gently the sample solution. Afterconˆrming that the temperature of the sample solution ismaintained at 20± 0.19C or at the temperature speciêd inthe monograph, measure the resistance RT (MQ) or conduc-tance GT (mS) of the sample solution, and calculate the con-ductivity kT by using the following equation.

kT＝ CGT or kT＝ C/RT

Items such as the sample preparation method, the necessi-ty of blank correction, the calculation method, the speciˆca-tion value, and the measuring temperature should be de-scribed in the monograph, if necessary.


Add the following:

73. Determination of Bulk andTapped Densities

Determination of bulk and tapped densities is a method todetermine the bulk densities of powdered drugs under looseand tapped packing conditions respectively. Loose packingis deˆned as the state obtained by pouring a powder sampleinto a vessel without any consolidation, and tapped packingis deˆned as the state obtained when the vessel containingthe powder sample is to be repeatedly dropped a speciêddistance at a constant drop rate until the apparent volume ofsample in the vessel becomes almost constant. The bulk den-sity is expressed in mass per unit apparent volume of powder(g/mL). Because the bulk density is one of the measures ofpacking properties, compressibility and ‰ow properties, andis dependent on the ``history'' of the powder, it is essentialto report the bulk density to specify how the determinationwas made.

Bulk densityThe bulk density is an apparent density obtained by pour-

ing a powder sample into a vessel without any consolidation.The determination of bulk density is achieved by measuringthe apparent volume of a powder sample having a knownmass in a graduated cylinder (Method 1) or by measuring themass of powder in a vessel having a known volume (Method2).

Method 1 (Constant mass method)Unless otherwise speciêd, pass a quantity of sample

su‹cient to complete the test through a 1000-mm (No.16)screen to break up agglomerates that may have formed dur-ing storage. Weigh accurately about 30 g of test sample, andpour it into a dry 100-mL graduated glass cylinder (readableto 1 mL). Carefully level the powder without consolidation,if necessary, and read the unsettled apparent volume, V0, tothe nearest graduated unit. Calculate the bulk density rB bythe formula:

rB＝MV0

rB: Bulk density by constant mass method (g/mL)M : Mass of powder sample (g)V0: Apparent volume of powder sample (mL)

Record the average of 3 determinations using 3 diŠerentpowder samples. If a 30-g sample is too large to determine,adjust the mass of sample so as to provide an apparentvolume of 60 – 100 mL.

Method 2 (Constant volume method)Unless otherwise speciêd, pass a quantity of sample

su‹cient to complete the test through a 1000-mm (No.16)screen to break up agglomerates that may have formed dur-ing storage. Allow an excess of sample powder to pour intothe measuring vessel having the volume of V and mass ofM0. Carefully scrape excess powder from the top of the ves-

sel by smoothly moving across it the edge of a slide glass orother tool. Remove any material from the sides of the vessel,and determine the total mass Mt. Calculate the bulk densityrB by the formula:

rB＝Mt－M0

VrB: Bulk density by constant volume method (g/mL)Mt: Total mass of powder and measuring vessel (g)M0: Mass of measuring vessel (g)V : Volume of measuring vessel (mL)

Record the average of 3 determinations using 3 diŠerentpowder samples.

Tapped densityTapped density is an apparent density obtained by

mechanically tapping a measuring vessel containing a pow-der sample. The determination of tapped density is achievedby measuring the apparent volume of a powder sample hav-ing a known mass in a vessel after tapping (Method 1) or bymeasuring the mass of powder in a vessel having a knownvolume after tapping (Method 2).

Method 1 (Constant mass method)Unless otherwise speciêd, pass a quantity of sample

su‹cient to complete the test through a 1000-mm (No.16) ora 710-mm (No.22) screen to break up agglomerates that mayhave formed during storage. Weigh accurately about 100 gof test sample, and pour it into a 250-mL graduated glasscylinder (readable to 2 mL) without consolidation. If it is notpossible to use 100 g, proceed according to the same proce-dure as that described above by using a 100-mL graduated g-lass cylinder (readable to 1 mL). It is essential to select ap-propriate masses of the cylinder support, holder and cylin-der so as to ensure the dynamic stability of the apparatusduring tapping. After attaching the glass cylinder containingthe powder sample to the tapping apparatus, carry out tap-ping under the measuring conditions (tapping rate and dropheight) speciêd for each apparatus.

Unless otherwise speciêd, repeat increments of 50 taps or1 minute until the diŠerence between succeeding measure-ments is less than 2z, and determine the ˆnal apparentvolume, Vf. Calculate the tapped density rT by the formula:

rT＝MVf

rT: Tapped density by constant mass method (g/mL)M : Mass of powder sample (g)Vf: Final apparent volume of sample after tapping (mL)

Record the average of 3 determinations using 3 diŠerentpowder samples.

Method 2 (Constant volume method)Unless otherwise speciêd, pass a quantity of sample

su‹cient to complete the test through a 1000-mm (No.16)screen to break up agglomerates that may have formed dur-ing storage. Attach a supplementary cylinder to the stainlesssteel measuring vessel having a known mass of M0 and avolume of V (Fig. 1), and then pour an excess of the sampleinto the vessel. After setting up the vessel in an adequate tap-


ping apparatus with a ˆxed drop height, carry out tapping atthe rate and cumulative tap number speciˆed for each ap-paratus. Then remove the supplementary cylinder from thevessel and carefully scrape excess powder from the top of thevessel by smoothly moving across it the edge of a slide glassor other tool. Remove any material from the sides of the ves-sel, and determine the total mass Mt. Calculate the tappeddensity rT by the formula:

rT＝Mt－M0

VrT: Tapped density by constant volume method (g/mL)

Mt: Total mass of powder and measuring vessel (g)M0: Mass of measuring vessel (g)V : Volume of measuring vessel (mL)

Record the average of 3 determinations and the relativestandard deviation using 3 diŠerent powder samples. If therelative standard deviation is not less than 2z, repeat thetest with further tapping.

Balances: Use balances readable to the nearest 0.1 g.

Fig. 1 Measuring vessel (left) and supplementary cylinder (right)

13771377

O‹cial Monographsfor Part I

Add the following:

Aceglutamide Aluminumアセグルタミドアルミニウム

C35H59Al3N10O24: 1084.84Pentakis[(2S )-2-acetylamino-4-carbamoylbutanoato]tetrahydroxotrialuminium[12607-92-0]

Aceglutamide Aluminum contains not less than85.4z and not more than 87.6z of aceglutamide(C7H12N2O4: 188.18), and not less than 7.0z and notmore than 8.0z of aluminum (Al: 26.98), calculatedon the dried basis.

Description Aceglutamide Aluminum occurs as a whitepowder, having astringent bitter taste.

It is freely soluble in water, and practically insoluble inethanol (99.5).

It dissolves in dilute hydrochloric acid.It is hygroscopic.

Identiˆcation (1) Dissolve 0.03 g each of AceglutamideAluminum and Aceglutamide Reference Standard in 5 mLof water, and use these solutions as the sample solution andthe standard solution. Perform the test with these solutionsas directed under the Thin-layer Chromatography. Spot5 mL each of the sample solution and the standard solutionon a plate of cellulose for thin-layer chromatography.Develop the plate with a mixture of 1-propanol, water andacetic acid (100) (16:8:1) to a distance of about 10 cm, andair-dry the plate. Spray evenly a solution of bromocresolgreen in ethanol (95) (1 in 1000), then spray evenly dilutedammonia solution (28) (1 in 100): the spots from the samplesolution and the standard solution show a light yellow andhave the same Rf value.

(2) A solution of Aceglutamide Aluminum in dilutehydrochloric acid (1 in 20) responds to the Qualitative Testsfor aluminum salt.

Optical rotation [a]20D :－5.5 –－7.59(2 g calculated on the

dried basis, water, 50 mL, 100 mm).

Purity (1) Heavy metals—Put 1.0 g of AceglutamideAluminum in a porcelain crucible, cover the crucible loosely,and heat gently to carbonize. After cooling, add 2 mL ofnitric acid and 1 mL of sulfuric acid, heat gently until thewhite fumes no more evolve, and heat to incinerate at 500 to6009C. If the incineration is not accomplished, add 2 mL ofnitric acid and 1 mL of sulfuric acid, heat in the same man-ner as above, then ignite at 500 to 6009C to incinerate. Aftercooling, add 2 mL of hydrochloric acid, proceed with thissolution according to Method 2, and perform the test. Pre-pare the control solution as follows: proceed in the samemanner as the preparation of the test solution with the sameamount of the reagents, and add 2.0 mL of Standard LeadSolution and water to make 50 mL (not more than 20 ppm).

(2) Arsenic—Prepare the test solution with 1.0 g ofAceglutamide Aluminum according to Method 1, and per-form the test (not more than 2 ppm).

(3) Related substances—Dissolve 0.10 g of Acegluta-mide Aluminum in the mobile phase to make exactly100 mL, and use this solution as the sample solution. Pipet1 mL of the sample solution, add the mobile phase to makeexactly 100 mL, and use this solution as the standardsolution (1). Separately, dissolve 10 mg of 2-acetamido-glutarimide in the mobile phase to make exactly 100 mL.Pipet 3 mL of this solution, add the mobile phase to makeexactly 100 mL, and use this solution as the standard solu-tion (2). Perform the test with exactly 20 mL each of thesample solution, the standard solution (1) and the standardsolution (2) as directed under the Liquid Chromatographyaccording to the following conditions, and determine eachpeak area by the automatic integration method: the peakarea of 2-acetamidoglutarimide from the sample solution isnot more than that from the standard solution (2), the peakareas other than aceglutamide and 2-acetamidoglutarimidefrom the sample solution are not more than 3/10 times thepeak area of aceglutamide from the standard solution (1),and the total of the peak areas other than aceglutamide and2-acetamidoglutarimide from the sample solution is notmore than the peak area of aceglutamide from the standardsolution (1).Operating conditions—

Detector, column, column temperature, mobile phase,and ‰ow rate: Proceed as directed in the operating condi-tions in the Assay.

Time span of measurement: About 3 times as long as theretention time of aceglutamide.System suitability—

Test for required detection: To exactly 5 mL of thestandard solution (1) add the mobile phase to make exactly

13781378 Supplement I, JP XIVO‹cial Monographs for Part I

50 mL. Conˆrm that the peak area of aceglutamide obtainedfrom 20 mL of this solution is equivalent to 7 to 13z of thatof aceglutamide obtained from 20 mL of the standard solu-tion.

System performance: Proceed as directed in the systemsuitability in the Assay.

System repeatability: When the test is repeated 6 timeswith 20 mL of the standard solution (1) under the aboveoperating conditions, the relative standard deviation of thepeak area of aceglutamide is not more than 2.0z.

Loss on drying Not more than 5.0z (1 g, 1309C, 5 hours).

Assay (1) Aceglutamide—Weigh accurately about 50 mgof Aceglutamide Aluminum, dissolve in a suitable amountof the mobile phase, add exactly 10 mL of the internal stan-dard solution and the mobile phase to make 50 mL, and usethis solution as the sample solution. Separately, weigh ac-curately about 45 mg of Aceglutamide Reference Standard,dissolve in a suitable amount of the mobile phase, add ex-actly 10 mL of the internal standard solution and the mobilephase to make 50 mL, and use this solution as the standardsolution. Perform the test with 10 mL each of the samplesolution and the standard solution as directed under the Liq-uid Chromatography according to the following conditions,and determine the ratios, QT and QS, of the peak area ofaceglutamide to that of the internal standard.

Amount (mg) of aceglutamide (C7H12N2O4)＝WS×QT

QS

WS: Amount (mg) of Aceglutamide Reference Standard

Internal standard solution—A solution of thymine inmethanol (1 in 4000).Operating conditions—

Detector: An ultraviolet absorption photometer (wave-length: 210 nm).

Column: A stainless steel column 4.6 mm in inside di-ameter and 25 cm in length, packed with octadecylsilanizedsilica gel for liquid chromatography (5 mm in particle di-ameter).

Column temperature: A constant temperature of about259C.

Mobile phase: A mixture of diluted perchloric acid (1 in1000) and methanol (99:1).

Flow rate: Adjust the ‰ow rate so that the retention timeof aceglutamide is about 5 minutes.System suitability—

System performance: When the procedure is run with10 mL of the standard solution under the above operatingconditions, aceglutamide and the internal standard are elut-ed in this order with the resolution between these peaksbeing not less than 11.

System repeatability: When the test is repeated 6 timeswith 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratiosof the peak area of aceglutamide to that of the internal stan-dard is not more than 1.0z.

(2) Aluminum—Weigh accurately about 3.0 g ofAceglutamide Aluminum, add 20 mL of dilute hydrochloric

acid, and heat on a water bath for 60 minutes. After cooling,add water to make exactly 200 mL. Pipet 20 mL of this solu-tion, add exactly 25 mL of 0.05 mol/L disodium dihydrogenethylenediamine tetraacetate VS and 20 mL of acetic acid-ammonium acetate buŠer solution, pH 4.8, and boil for5 minutes. After cooling, add 50 mL of ethanol (95), andtitrate with 0.05 mol/L zinc acetate VS until the color of thesolution changes from light dark green to light red (indica-tor: 2 mL of dithizone TS). Perform a blank determinationin the same manner.

Each mL of 0.05 mol/L disodium dihydrogenethylenediamine tetraacetate VS＝ 1.349 mg of Al

Containers and storage Containers—Tight containers.

Acetylkitasamycinアセチルキタサマイシン

Change the Assay (3) to read:

Assay(3) Standard solution－Weigh accurately an amount of

Leucomycin A5 Reference Standard equivalent to about30 mg (potency), dissolve in 10 mL of methanol, add waterto make exactly 100 mL, and use this solution as the stan-dard stock solution. Keep the standard stock solution at 59Cor below and use within 3 days. Take exactly a suitableamount of the standard stock solution before use, add0.1 mol/L phosphate buŠer solution, pH 8.0 to make solu-tions so that each mL contains 30 mg (potency) and 7.5 mg(potency), and use these solutions as the high concentrationstandard solution and the low concentration standard solu-tion, respectively.

Acetylspiramycinアセチルスピラマイシン

Change to read except the structural formulaand chemical name:

Acetylspiramycin contains not less than 900 mg(potency) per mg, calculated on the dried basis. Thepotency of Acetylspiramycin is expressed as mass(potency) of acetylspiramycin II (C47H78N2O16:927.13). 1 mg (potency) of Acetylspiramycin isequivalent to 0.7225 mg of acetylspiramycin II(C47H78N2O16).

Description Acetylspiramycin occurs as a white to lightyellowish white powder.

It is very soluble in acetonitrile and in methanol, freelysoluble in ethanol (99.5), and practically insoluble in water.

Identiˆcation (1) Determine the absorption spectrum ofa solution of Acetylspiramycin in methanol (1 in 50,000) as

13791379Supplement I, JP XIV O‹cial Monographs for Part I

directed under the Ultraviolet-visible Spectrophotometry,and compare the spectrum with the Reference Spectrum:both spectra exhibit similar intensities of absorption at thesame wavelengths.

(2) Determine the infrared absorption spectrum ofAcetylspiramycin as directed in the potassium bromide diskmethod under the Infrared Spectrophotometry, and com-pare the spectrum with the Reference Spectrum: both spec-tra exhibit similar intensities of absorption at the same wavenumbers.

Content ratio of the active principle Dissolve 25 mg ofAcetylspiramycin in 25 mL of the mobile phase, and use thissolution as the sample solution. Perform the test with 5 mLof the sample solution as directed under the Liquid Chro-matography according to the following conditions, anddetermine the areas, AII, AIII, AIV, AV, AVI and AVII, of thepeaks of acetylspiramycin II, acetylspiramycin III, acetyl-spiramycin IV, acetylspiramycin V, acetylspiramycin VI andacetylspiramycin VII, respectively, by the automatic integra-tion method, and calculate the ratios of the amounts of AII,AIV and the total of AIII and AV to the total amount of allthese peaks: the amount of AII is 30 – 45z, AIV is 30 – 45z,and the total of AIII and AV is not more than 25z. The rela-tive retention times of acetylspiramycin III, acetylspiramy-cin IV, acetylspiramycin V, acetylspiramycin VI and acetyl-spiramycin VII with respect to acetylspiramycin II are 1.3,1.7, 2.3, 0.85 and 1.4, respectively.Operating conditions—


Column: A stainless steel column 6 mm in inside diameterand 15 cm in length, packed with octadecylsilanized silica gelfor liquid chromatography (3 mm in particle diameter).


Mobile phase: A mixture of acetonitrile, 0.02 mol/Lpotassium dihydrogen phosphate TS and a solution ofdipotassium hydrogen phosphate (87 in 25,000) (26:7:7).

Flow rate: Adjust the ‰ow rate so that the retention timeof acetylspiramycin II is about 10 minutes.System suitability—

System performance: Dissolve 25 mg of AcetylspiramycinII Reference Standard in the mobile phase to make 100 mL.When the procedure is run with 5 mL of this solution underthe above operating conditions, the number of theoreticalplates and the symmetry coe‹cient of the peak of acetyl-spiramycin II are not less than 14,500 and not more than 2.0,respectively.

System repeatability: When the test is repeated 6 timeswith 5 mL of the sample solution under the above operatingconditions, the relative standard deviation of the peak areaof acetylspiramycin II is not more than 2.0z.

Purity (1) Heavy metals—Proceed with 1.0 g of Acetyl-spiramycin according to Method 2, and perform the test.Prepare the control solution with 1.0 mL of Standard LeadSolution (not more than 10 ppm).

(2) Arsenic—Prepare the test solution with 2.0 g ofAcetylspiramycin according to Method 3, and perform thetest (not more than 1 ppm).

Loss on drying Not more than 3.0z (1 g, in vacuum,phosphorus (V) oxide, 609C, 3 hours).

Residue on ignition Not more than 0.50z (1.0 g).

Assay Perform the test according to the Cylinder-platemethod as directed under the Microbial Assay for Antibiot-ics according to the following conditions.

(1) Test organism—Bacillus subtilis ATCC 6633(2) Culture medium—Use the medium i in 1) Medium

for test organism [5] under (1) Agar media for seed and baselayer.

(3) Standard solutions—Weigh accurately an amountof Acetylspiramycin II Reference Standard, equivalent toabout 50 mg (potency), dissolve in 20 mL of methanol, add0.1 mol/L phosphate buŠer solution for antibiotics, pH 8.0to make exactly 50 mL, and use this solution as the standardstock solution. Keep the standard stock solution at not ex-ceeding 59C, and use within 3 days. Take exactly a suitableamount of the standard stock solution before use, add0.1 mol/L phosphate buŠer solution for antibiotics, pH 8.0to make solutions so that each mL contains 80 mg (potency)and 20 mg (potency), and use these solutions as the high con-centration standard solution and the low concentration stan-dard solution, respectively.

(4) Sample solutions—Weigh accurately an amount ofAcetylspiramycin, equivalent to about 50 mg (potency), dis-solve in 20 mL of methanol, and add 0.1 mol/L phosphatebuŠer solution for antibiotics, pH 8.0 to make exactly50 mL. Take exactly a suitable amount of this solution, add0.1 mol/L phosphate buŠer solution for antibiotics, pH 8.0to make solutions so that each mL contains 80 mg (potency)and 20 mg (potency), and use these solutions as the high con-centration sample solution and the low concentration samplesolution, respectively.


Aclarubicin Hydrochloride塩酸アクラルビシン


Aclarubicin Hydrochloride contains not less than860 mg (potency) per mg, calculated on the anhydrousbasis. The potency of Aclarubicin Hydrochlorideis expressed as mass (potency) of aclarubicin(C42H53NO15: 811.87).

Description Aclarubicin Hydrochloride occurs as a yellowto pale orange-yellow powder.

It is very soluble in chloroform and in methanol, freelysoluble in water, and slightly soluble in ethanol (95).


Identiˆcation (1) Determine the absorption spectrum ofa solution of Aclarubicin Hydrochloride in diluted methanol(4 in 5) (3 in 100,000) as directed under the Ultraviolet-visible Spectrophotometry, and compare the spectrum withthe Reference Spectrum: both spectra exhibit similar intensi-ties of absorption at the same wavelengths.

(2) Determine the infrared absorption spectrum ofAclarubicin Hydrochloride as directed in the potassiumbromide disk method under the Infrared Spectrophotomet-ry, and compare the spectrum with the Reference Spectrum:both spectra exhibit similar intensities of absorption at thesame wave numbers.

(3) A solution of Aclarubicin Hydrochloride inmethanol (1 in 200) responds to the Qualitative Test (2) forchloride.

Optical rotation [a]20D : －146 –－1629(0.05 g calculated on

the anhydrous basis, water, 10 mL, 100 mm).

pH The pH of a solution obtained by dissolving 0.05 g ofAclarubicin Hydrochloride in 10 mL of water is between 5.5and 6.5.

Purity (1) Clarity and color of solution－Dissolve 0.10 gof Aclarubicin Hydrochloride in 10 mL of water: the solu-tion is clear and yellow to pale orange-yellow.

(2) Heavy metals—Proceed with 1.0 g of AclarubicinHydrochloride according to Method 2, and perform the test.Prepare the control solution with 2.0 mL of Standard LeadSolution (not more than 20 ppm).

(3) Related substances—Dissolve 10 mg of AclarubicinHydrochloride in 10 mL of the mobile phase, and use thissolution as the sample solution. Perform the test with 20 mLof the sample solution as directed under the Liquid Chro-matography according to the following conditions, anddetermine each peak area by the automatic integrationmethod. Calculate the amount of the related substances bythe area percentage method: the amount of aklavinone hav-ing the relative retention time of about 0.6 to aclarubicin isnot more than 0.2z, aclacinomycin L1 having the relativeretention time of about 0.75 to aclarubicin is not more than0.5z, 1-deoxypyrromycin having the relative retention timeof about 1.7 to aclarubicin is not more than 1.5z andaclacinomycin S1 having the relative retention time of about2.3 to aclarubicin is not more than 0.5z, and the totalamount of the peaks other than aclarubicin and the peaksmentioned above is not more than 1.0z of the peak area ofaclarubicin.Operating conditions—

Detector: A visible absorption photometer (wavelength:436 nm).

Column: A stainless steel column 3.9 mm in inside di-ameter and 30 cm in length, packed with silica gel for liquidchromatography (10 mm in particle diameter).


Mobile phase: A mixture of chloroform, methanol, aceticacid (100), water and triethylamine (6800:2000:1000:200:1).

Flow rate: Adjust the ‰ow rate so that the retention timeof aclarubicin is about 5 minutes.

Time span of measurement: As long as about 4 times ofthe retention time of aclarubicin after the solvent peak.System suitability—

Test for required detectability: Pipet 1 mL of the samplesolution, add the mobile phase to make exactly 100 mL, anduse this solution as the solution for system suitability test.Pipet 1 mL of the solution for system suitability test, andadd the mobile phase to make exactly 10 mL. Conˆrm thatthe peak area of aclarubicin obtained from 20 mL of this so-lution is equivalent to 7 to 13z of that from 20 mL of the so-lution for system suitability test.

System performance: Dissolve 5 mg of AclarubicinHydrochloride in 10 mL of 0.1 mol/L hydrochloric acid TS,and allow to stand for 60 minutes. To 1.0 mL of this solu-tion add 1.0 mL of 0.2 mol/L sodium hydroxide TS, 1.0 mLof phosphate buŠer solution, pH 8.0 and 1.0 mL of chlo-roform, shake vigorously, and take the chloroform layer.When the procedure is run with 20 mL of the chloroformunder the above operating conditions, aclarubicin and 1-deoxypyrromycin are eluted in this order with the resolutionbetween these peaks being not less than 3.0.

System repeatability: When the test is repeated 5 timeswith 20 mL of the sample solution under the above operatingconditions, the relative standard deviation of the peak areaof aclarubicin is not more than 2.0z.

Water Not more than 3.5z (0.1 g, volumetric titration,direct titration).

Residue on ignition Not more than 0.1z (1 g).

Assay Weigh accurately an amount of AclarubicinHydrochloride, equivalent to about 20 mg (potency), anddissolve in diluted methanol (4 in 5) to make exactly 100 mL.Pipet 15 mL of this solution, add diluted methanol (4 in 5) tomake exactly 100 mL, and use this solution as the samplesolution. Separately, weigh accurately an amount ofAclarubicin Reference Standard, equivalent to about 20 mg(potency), add 0.6 mL of diluted hydrochloric acid (1 in 250)and diluted methanol (4 in 5) to make exactly 100 mL. Pipet15 mL of this solution, add diluted methanol (4 in 5) to makeexactly 100 mL, and use this solution as the standard solu-tion. Perform the test with the sample solution and the stan-dard solution as directed under the Ultraviolet-visible Spec-trophotometry, and determine the absorbencies, AT and AS,at 433 nm.

Amount [mg (potency)] of aclarubicin (C42H53NO15)

＝WS×AT

AS× 1000

WS: Amount [mg (potency)] of Aclarubicin ReferenceStandard

Containers and storage Containers—Tight containers.Storage—Light-resistant and at 59C or below.


Change to read:

Actinomycin DアクチノマイシンD

C62H86N12O16: 1255.42[50-76-0]

Actinomycin D, when dried, contains not less than950 mg (potency) per mg. The potency of ActinomycinD is expressed as mass (potency) of actinomycin D(C62H86N12O16).

Description Actinomycin D occurs as an orange-red to redcrystalline powder.

It is freely soluble in acetone, sparingly soluble in acetoni-trile and in methanol, slightly soluble in ethanol (99.5), andvery slightly soluble in water.

Identiˆcation (1) Determine the absorption spectrum ofa solution of Actinomycin D in methanol (3 in 100,000) asdirected under the Ultraviolet-visible Spectrophotometry,and compare the spectrum with the Reference Spectrum orthe spectrum of a solution of Actinomycin D ReferenceStandard prepared in the same manner as the sample solu-tion: both spectra exhibit similar intensities of absorption atthe same wavelengths.

(2) Dissolve 0.1 g each of Actinomycin D and Actinomy-cin D Reference Standard in 10 mL of acetone, and use thesesolutions as the sample solution and the standard solution.Perform the test with these solutions as directed under theThin-layer Chromatography. Spot 10 mL each of the samplesolution and the standard solution on a plate of silica gelwith ‰uorescent indicator for thin-layer chromatography.Develop the plate with a mixture of 1-butanol, water andmethanol (4:2:1) to a distance of about 10 cm, and air-drythe plate. Examine under ultraviolet light (main wavelength:254 nm): the Rf value of the principal spot from the samplesolution is the same as that from the standard solution.

Optical rotation [a]20D : －292 –－3179(after drying, 10 mg,

methanol, 10 mL, 100 mm).

Loss on drying Not more than 5.0z (1 g, in vacuum,609C, 3 hours).

Assay Weigh accurately an amount of Actinomycin Dand Actinomycin D Reference Standard, previously dried,equivalent to about 60 mg (potency), dissolve each in themobile phase to make exactly 50 mL, and use these solutionsas the sample solution and the standard solution. Perform

the test with exactly 25 mL each of the sample solution andthe standard solution as directed under the Liquid Chro-matography according to the following conditions, anddetermine the peak area of actinomycin D, AT and AS, ofboth solutions.

Amount [mg (potency)] of C62H86N12O16

＝WS×AT

AS× 1000

WS: Amount [mg (potency)] of Actinomycin D ReferenceStandard

Operating conditions—Detector: An ultraviolet absorption photometer (wave-

length: 254 nm).Column: A stainless steel column 3.9 mm in inside di-

ameter and 30 cm in length, packed with octadecylsilanizedsilica gel for liquid chromatography (10 mm in particlediameter).


Mobile phase: A mixture of 0.02 mol/L acetic acid-sodium acetate TS and acetonitrile (25:23).

Flow rate: Adjust the ‰ow rate so that the retention timeof actinomycin D is about 23 minutes.System suitability—

System performance: When the procedure is run with25 mL of the standard solution under the above operatingconditions, the number of theoretical plates and the sym-metrical coe‹cient of the peak of actinomycin D are not lessthan 2000 steps and not more than 1.5, respectively.

System repeatability: When the test is repeated 5 timeswith 25 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peakarea of actinomycin D is not more than 2.0z.

Containers and storage Containers—Tight containers.Storage—Light-resistant.

Amoxicillinアモキシシリン

Change the origin/limits of content to read:

Amoxicillin contains not less than 950 mg (potency)per mg, calculated on the anhydrous basis. The poten-cy of Amoxicillin is expressed as mass (potency) ofamoxicillin (C16H19N3O5S: 365.40).

Add the following next to Identiˆcation:

Optical rotation [a]20D : ＋290 –＋3159(0.1 g calculated on


Add the following next to Purity (2):

Purity(3) Related substances—Dissolve 0.10 g of Amoxicillin


in 50 mL of a solution of sodium tetraborate decahydrate(1 in 200), and use this solution as the sample solution. Pipet1 mL of the sample solution, add a solution of sodiumtetraborate decahydrate (1 in 200) to make exactly 100 mL,and use this solution as the standard solution. Perform thetest with exactly 10 mL each of the sample solution and thestandard solution as directed under the Liquid Chro-matography according to the following conditions, anddetermine each peak area by the automatic integrationmethod: the area of the peak other than amoxicillin obtainedfrom the sample solution is not more than the peak area ofamoxicillin from the standard solution.Operating conditions—




Mobile phase: Dissolve 1.361 g of sodium acetate trihy-drate in 750 mL of water, adjust the pH to 4.5 with aceticacid (31), and add water to make 1000 mL. To 950 mL ofthis solution add 50 mL of methanol.

Flow rate: Adjust the ‰ow rate so that the retention timeof amoxicillin is about 8 minutes.

Time span of measurement: About 4 times as long as theretention time of amoxicillin.System suitability—

Test for required detection: To exactly 1 mL of the stan-dard solution add a solution of sodium tetraborate decahy-drate (1 in 200) to make exactly 10 mL. Conˆrm that thepeak area of amoxicillin obtained from 10 mL of this solu-tion is equivalent to 7 to 13z of that of amoxicillin obtainedfrom 10 mL of the standard solution.

System performance: When the procedure is run with10 mL of the standard solution under the above operatingconditions, the number of theoretical plates of the peak ofamoxicillin is not less than 2500.

System repeatability: When the test is repeated 6 timeswith 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peakarea of amoxicillin is not more than 1.0z.

Change the Assay to read:

Assay Weigh accurately an amount of Amoxicillin andAmoxicillin Reference Standard, equivalent to about 30 mg(potency), dissolve each in a solution of boric acid (1 in 200)to make exactly 100 mL, and use these solutions as the sam-ple solution and the standard solution. Perform the test withexactly 10 mL each of the sample solution and the standardsolution as directed under the Liquid Chromatography ac-cording to the following conditions, and calculate the peakareas, AT and AS, of amoxicillin of each solution.

Amount [mg (potency)] of C16H19N3O5S

＝WS×AT

AS× 1000

WS: Amount [mg (potency)] of Amoxicillin ReferenceStandard





Mobile phase: Dissolve 1.361 g of sodium acetate trihy-drate in 750 mL of water, adjust the pH to 4.5 with aceticacid (31), and add water to make 1000 mL. To 950 mL ofthis solution add 50 mL of methanol.

Flow rate: Adjust the ‰ow rate so that the retention timeof amoxicillin is about 8 minutes.System suitability—

System performance: When the procedure is run with10 mL of the standard solution under the above operatingconditions, the number of theoretical plates of the peak ofamoxicillin is not less than 2500.

System repeatability: When the test is repeated 6 timeswith 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peakarea of amoxicillin is not more than 1.0z.

Ampicillinアンピシリン


Ampicillin contains not less than 960 mg (potency)per mg, calculated on the anhydrous basis. The poten-cy of Ampicillin is expressed as mass (potency) ofampicillin (C16H19N3O4S: 349.40).

Description Ampicillin occurs as a white to light yellowishwhite, crystals or crystalline powder.

It is sparingly soluble in water, slightly soluble inmethanol, very slightly soluble in ethanol (95), and practical-ly insoluble in acetonitrile.

Identiˆcation Determine the infrared absorption spectrumof Ampicillin as directed in the potassium bromide diskmethod under the Infrared Spectrophotometry, and com-pare the spectrum with the Reference Spectrum or the spec-trum of Ampicillin Reference Standard: both spectra exhibitsimilar intensities of absorption at the same wave numbers.



pH The pH of a solution obtained by dissolving 1.0 g ofAmpicillin in 400 mL of water is between 3.5 and 5.5.

Purity (1) Heavy metals—Proceed with 1.0 g of Ampicil-


lin according to Method 2, and perform the test. Prepare thecontrol solution with 2.0 mL of Standard Lead Solution (notmore than 20 ppm).

(2) Arsenic—Prepare the test solution with 1.0 g ofAmpicillin according to Method 3, and perform the test (notmore than 2 ppm).

(3) Related substances—Dissolve 50 mg of Ampicillin inthe mobile phase to make 50 mL, and use this solution as thesample solution. Pipet 1 mL of the sample solution, add themobile phase to make exactly 100 mL, and use this solutionas the standard solution. Perform the test with exactly 10 mLeach of the sample solution and the standard solution asdirected under the Liquid Chromatography according to thefollowing conditions, and determine each peak area by theautomatic integration method: each peak area other thanthat of ampicillin obtained from the sample solution is notmore than the peak area of ampicillin from the standardsolution.Operating conditions—


Time span of measurement: About 10 times as long as theretention time of ampicillin.System suitability—

System performance, and system repeatability: Proceed asdirected in the system suitability in the Assay.

Test for required detectability: Measure exactly 1 mL ofthe standard solution, and add the mobile phase to makeexactly 10 mL. Conˆrm that the peak area of ampicillinobtained from 10 mL of this solution is equivalent to 7 to13z of that from 10 mL of the standard solution.

(4) N,N-Dimethylaniline—Weigh accurately about 1 gof Ampicillin, dissolve in 5 mL of sodium hydroxide TS,add exactly 1 mL of the internal standard solution, shakevigorously for 1 minute, and use the upper layer liquidobtained after allowing it to stand as the sample solution.Separately, weigh accurately about 50 mg of N,N-dimethylaniline, dissolve in 2 mL of hydrochloric acid and20 mL of water, add water to make exactly 50 mL, and usethis solution as the standard stock solution. Pipet 5 mL ofthe standard stock solution, and add water to make exactly250 mL. Pipet 1 mL of this solution, add 5 mL of sodiumhydroxide TS and exactly 1 mL of the internal standardsolution, shake vigorously for 1 minute, and use the upperlayer liquid obtained after allowing it to stand as the stan-dard solution. Perform the test with 1 mL each of the samplesolution and the standard solution as directed under the GasChromatography according to the following conditions,determine the ratios, QT and QS, of the peak area of N,N-dimethylaniline to that of the internal standard, and calcu-late the amount of N,N-dimethylaniline by the followingequation: not more than 20 ppm.

Amount (ppm) of N,N-dimethylaniline＝WS

WT×

QT

QS× 400

WS: Amount (g) of N,N-dimethylanilineWT: Amount (g) of the sample

Internal standard solution—A solution of naphthalene incyclohexane (1 in 20,000).Operating conditions—

Detector: A hydrogen ‰ame-ionization detector.Column: A glass column 2.6 mm in inside diameter and

2 m in length, packed with siliceous earth for gas chro-matography (180 – 250 mm in particle diameter) coated with50z phenyl-50z methyl polysiloxane for gas chro-matography at the ratio of 3z.


Carrier gas: HeliumFlow rate: Adjust the ‰ow rate so that the retention time

of N,N-dimethylaniline is about 5 minutes.System suitability—

Test for required detectability: Measure exactly 1 mL ofthe standard stock solution, and add water to make exactly250 mL. Pipet 1 mL of this solution, add 5 mL of sodiumhydroxide TS and exactly 1 mL of the internal standardsolution, shake vigorously for 1 minute, and use the upperlayer liquid obtained after allowing it to stand for the test.Conˆrm that when the procedure is run with 1 mL of theupper layer liquid under the above operating conditions, theratio of the peak area of N,N-dimethylaniline to that of theinternal standard is equivalent to 15 – 25z of the ratio ofthe peak area of N,N-dimethylaniline to that of the internalstandard obtained from the standard solution.

System performance: Dissolve 50 mg of N,N-dimethylani-line in cyclohexane to make 50 mL. To 1 mL of this solutionadd the internal standard solution to make 50 mL, and usethis solution as the solution for system performance test.When the procedure is run with 1 mL of the solution for sys-tem performance test under the above operating conditions,N,N-dimethylaniline and the internal standard are eluted inthis order with the resolution between these peaks being notless than 3.

System repeatability: When the test is repeated 6 timeswith 1 mL of the solution for system performance test underthe above operating conditions, the relative standard devia-tion of the ratios of the peak area of N,N-dimethylaniline tothat of the internal standard is not more than 2.0z.

Water 12.0 – 15.0z (0.1 g, volumetric titration, directtitration).

Assay Weigh accurately an amount of Ampicillin andAmpicillin Reference Standard, equivalent to about 50 mg(potency), dissolve in a suitable volume of the mobile phase,add exactly 5 mL each of the internal standard solution andthe mobile phase to make 50 mL, and use these solutions asthe sample solution and the standard solution. Perform thetest with 10 mL each of the sample solution and the standardsolution as directed under the Liquid Chromatographyaccording to the following conditions, and determine theratios, QT and QS, of the peak area of ampicillin to that ofthe internal standard.

Amount [mg (potency)] of C16H19N3O4S＝WS×QT

QS× 1000

WS: Amount [mg (potency)] of Ampicillin Reference


Standard

Internal standard solution—A solution of guaifenesin in themobile phase (1 in 200).Operating conditions—


Column: A stainless steel column 4.6 mm in inside di-ameter and 15 cm in length, packed with octadecylsilanizedsilica gel for liquid chromatography (5 mm in particlediameter).


Mobile phase: Dissolve 5.94 g of diammonium hydrogenphosphate in 850 mL of water, add 100 mL of acetonitrile,adjust the pH to 5.0 with phosphoric acid, and add water tomake exactly 1000 mL.

Flow rate: Adjust the ‰ow rate so that the retention timeof ampicillin is about 6 minutes.System suitability—

System performance: When the procedure is run with10 mL of the standard solution under the above operatingconditions, ampicillin and the internal standard are eluted inthis order with the resolution between these peaks being notless than 40.

System repeatability: When the test is repeated 6 timeswith 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratiosof the peak area of ampicillin to that of the internal standardis not more than 1.0z.


Anhydrous Ampicillin無水アンピシリン


Anhydrous Ampicillin contains not less than 960 mg(potency) per mg, calculated on the anhydrous basis.The potency of Anhydrous Ampicillin is expressed asmass (potency) of ampicillin (C16H19N3O4S).

Description Anhydrous Ampicillin occurs as white to lightyellowish white, crystals or crystalline powder. It is sparinglysoluble in water, slightly soluble in methanol, very slightlysoluble in ethanol (95), and practically insoluble in acetoni-trile.

Identiˆcation Determine the infrared absorption spectrumof Anhydrous Ampicillin as directed in the potassiumbromide disk method under the Infrared Spectrophotomet-ry, and compare the spectrum with the Reference Spectrum:both spectra exhibit similar intensities of absorption at thesame wave numbers.



pH The pH of a solution obtained by dissolving 1.0 g ofAnhydrous Ampicillin in 100 mL of water is between 4.0 and5.5.

Purity (1) Heavy metals—Proceed with 1.0 g of Anhy-drous Ampicillin according to Method 2, and perform thetest. Prepare the control solution with 2.0 mL of StandardLead Solution (not more than 20 ppm).

(2) Arsenic—Prepare the test solution with 1.0 g of An-hydrous Ampicillin according to Method 3, and perform thetest (not more than 2 ppm).

(3) Related substances—Dissolve 0.05 g of AnhydrousAmpicillin in the mobile phase to make 50 mL, and use thissolution as the sample solution. Pipet 1 mL of the samplesolution, add the mobile phase to make exactly 100 mL, anduse this solution as the standard solution. Perform the testwith exactly 10 mL each of the sample solution and the stan-dard solution as directed under the Liquid Chromatographyaccording to the following conditions, and determine eachpeak area by the automatic integration method: the area ofeach peak other than ampicillin from the sample solution isnot more than the peak area of ampicillin from the standardsolution.Operating conditions—


Time span of measurement: As long as about 10 times ofthe retention time of ampicillin.System suitability—

Test for required detectability: To exactly 1 mL of thestandard solution add the mobile phase to make exactly10 mL. Conˆrm that the peak area of ampicillin obtainedfrom 10 mL of this solution is equivalent to 7 to 13z of thatfrom 10 mL of the standard solution.



Assay Weigh accurately an amount of Anhydrous Am-picillin and Ampicillin Reference Standard, equivalent toabout 50 mg (potency), add exactly 5 mL each of the internalstandard solution and the mobile phase to make 50 mL, anduse these solutions as the sample solution and the standardsolution. Perform the test with 10 mL each of the samplesolution and the standard solution as directed under theLiquid Chromatography according to the following condi-tions, and determine the ratios, QT and QS, of the peak areaof ampicillin to that of the internal standard.

Amount [mg (potency)] of ampicillin (C16H19N3O4S)

＝WS×QT

QS× 1000

WS: Amount [mg (potency)] of Ampicillin ReferenceStandard

Internal standard solution—A solution of guaifenesin in the


mobile phase (1 in 200).Operating conditions—






System performance: When the procedure is run with10 mL of the standard solution under the above operatingconditions, ampicillin and the internal standard are eluted inthis order with the resolution between these peaks being notless than 40.

System repeatability: When the test is repeated 6 timeswith 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratiosof the peak area of ampicillin to that of the internal standardis not more than 1z.


Ampicillin Sodiumアンピシリンナトリウム


Ampicillin Sodium contains not less than 850 mg(potency) per mg, calculated on the anhydrous basis.The potency of Ampicillin Sodium is expressed asmass (potency) of ampicillin (C16H19N3O4S: 349.40).

Description Ampicillin Sodium occurs as white to light yel-lowish white, crystals or crystalline powder.

It is very soluble in water, and sparingly soluble in ethanol(99.5).

Identiˆcation (1) Determine the infrared absorptionspectrum of Ampicillin Sodium, previously dried in a desic-cator (reduced pressure not exceeding 0.67 kPa, 609C) for3 hours, as directed in the potassium bromide disk methodunder the Infrared Spectrophotometry, and compare thespectrum with the Reference Spectrum: both spectra exhibitsimilar intensities of absorption at the same wave numbers.

(2) Ampicillin Sodium responds to the Qualitative Test(1) for sodium salt.

Optical rotation [a]20D : ＋246 –＋2729(1 g calculated on the

anhydrous basis, water, 100 mL, 100 mm).

pH The pH of a solution obtained by dissolving 1.0 g ofAmpicillin Sodium in 10 mL of water is between 8.0 and10.0.

Purity (1) Clarity and color of solution—Dissolve 1.0 gof Ampicillin Sodium in 10 mL of water: the solution is clearand colorless or pale yellow.

(2) Heavy metals—Proceed with 1.0 g of AmpicillinSodium according to Method 1, and perform the test.Prepare the control solution with 2.0 mL of Standard LeadSolution (not more than 20 ppm).

(3) Arsenic—Prepare the test solution with 1.0 g ofAmpicillin Sodium according to Method 1, and perform thetest (not more than 2 ppm).

(4) Related substances—Dissolve 50 mg of AmpicillinSodium in 50 mL of the mobile phase, and use this solutionas the sample solution. Pipet 1 mL of the sample solution,add the mobile phase to make exactly 100 mL, and use thissolution as the standard solution. Perform the test withexactly 10 mL each of the sample solution and the standardsolution as directed under the Liquid Chromatographyaccording to the following conditions, and determine eachpeak area by the automatic integration method: the peakarea other than ampicillin obtained from the sample solutionis not more than the peak area of ampicillin from the stan-dard solution.Operating conditions—





Flow rate: Adjust the ‰ow rate so that the retention timeof ampicillin is about 6 minutes.

Time span of measurement: About 10 times as long as theretention time of ampicillin.System suitability—

Test for required detectability: Measure exactly 1 mL ofthe standard solution, and add the mobile phase to makeexactly 10 mL. Conˆrm that the peak area of ampicillinobtained from 10 mL of this solution is equivalent to 7 to13z of that of ampicillin obtained from 10 mL of the stan-dard solution.

System performance: Dissolve 50 mg of AmpicillinReference Standard in a suitable amount of the mobilephase, add 5 mL of a solution of guaifenesin in the mobilephase (1 in 200) and the mobile phase to make 50 mL, anduse this solution as the solution for system suitability test.When the procedure is run with 10 mL of the solution for sys-tem suitability test under the above operating conditions,ampicillin and guaifenesin are eluted in this order with the


resolution between these peaks being not less than 35.System repeatability: When the test is repeated 6 times

with 10 mL of the solution for system suitability test underthe above operating conditions, the relative standard devia-tion of the ratios of the peak area of ampicillin to that ofguaifenesin is not more than 1.0z.




for test organism [5] under (1) Agar media for seed and baselayer, having pH 6.5 to 6.6 after sterilization.

(3) Standard solutions—Weigh accurately an amount ofAmpicillin Reference Standard, equivalent to about 25 mg(potency), and dissolve in phosphate buŠer solution, pH 6.0to make exactly 50 mL. Take exactly a suitable amount ofthis solution, add phosphate buŠer solution, pH 6.0 to makesolutions so that each mL contains 5 mg (potency) and1.25 mg (potency), and use these solutions as the high con-centration standard solution and the low concentration stan-dard solution, respectively.

(4) Sample solutions—Weigh accurately an amount ofAmpicillin Sodium, equivalent to about 25 mg (potency),and dissolve in phosphate buŠer solution, pH 6.0 to makeexactly 50 mL. Take exactly a suitable amount of this solu-tion, add phosphate buŠer solution, pH 6.0 to make solu-tions so that each mL contains 5 mg (potency) and 1.25 mg(potency), and use these solutions as the high concentrationsample solution and the low concentration sample solution,respectively.


Arbekacin Sulfate硫酸アルベカシン


Arbekacin Sulfate contains not less than 670 mg(potency) per mg, calculated on the dried basis. Thepotency of Arbekacin Sulfate is expressed as mass(potency) of arbekacin (C22H44N6O10: 552.62).

Description Arbekacin Sulfate occurs as a white powder.It is very soluble in water, and practically insoluble in

ethanol (99.5).

Identiˆcation (1) Dissolve 10 mg each of Arbekacin Sul-fate and Arbekacin Sulfate Reference Standard in 1 mL ofwater, and use these solutions as the sample solution and thestandard solution. Perform the test with these solutions asdirected under the Thin-layer Chromatography. Spot 2 mL

each of the sample solution and the standard solution on aplate of silica gel for thin-layer chromatography. Developthe plate with a mixture of ammonia solution (28),methanol, chloroform and ethanol (95) (7:6:4:1) to a dis-tance of about 10 cm, and air-dry the plate. Spray evenly0.2z ninhydrin-water saturated 1-butanol TS on the plate,and heat at 1009C for 10 minutes: the principal spot ob-tained from the sample solution and the spot from the stan-dard solution are purple-brown in color and their Rf valuesare the same.

(2) A solution of Arbekacin Sulfate (1 in 50) responds tothe Qualitative Test (1) for sulfate.

Optical rotation [a]20D : ＋69 –＋799(0.25 g after drying,

water, 25 mL, 100 mm).

pH The pH of a solution obtained by dissolving 0.75 g ofArbekacin Sulfate in 10 mL of water is between 6.0 and 8.0.

Purity (1) Clarity and color of solution—A solutionobtained by dissolving 1.0 g of Arbekacin Sulfate in 5 mL ofwater is clear and colorless.

(2) Heavy metals—Proceed with 2.0 g of Arbekacin Sul-fate according to Method 1, and perform the test. Preparethe control solution with 2.0 mL of Standard Lead Solution(not more than 10 ppm).

(3) Dibekacin—Weigh accurately about 20 mg ofArbekacin Sulfate, add exactly 10 mL of the internal stan-dard solution to dissolve, add water to make 20 mL, and usethis solution as the sample solution. Separately, weighaccurately an amount of Dibekacin Sulfate Reference Stan-dard, equivalent to about 10 mg (potency), and dissolve inwater to make exactly 50 mL. Pipet 5 mL of this solution,add exactly 10 mL of the internal standard solution andwater to make 20 mL, and use this solution as the standardsolution. Perform the test with 5 mL each of the sample solu-tion and the standard solution as directed under the LiquidChromatography according to the following conditions, anddetermine the ratios, QT and QS, of the peak area of dibeka-cin to that of the internal standard. Calculate the amount ofdibekacin by the following equation: not more than 2.0z.

Amount (z) of dibekacin＝WS

WT×

QT

QS×

150× 1000

WS: Amount [mg (potency)] of Dibekacin Sulfate Refer-ence Standard

WT: Amount (mg) of the sample

Internal standard solution—A solution of bekanamycin sul-fate (1 in 2000).Operating conditions—

Detector: Fluorometry (excitation wavelength: 340 nm,detection wavelength: 460 nm).



Reaction coil: A column about 0.3 mm in inside diameter


and about 3 m in length.Reaction coil temperature: A constant temperature of

about 509C.Mobile phase: Dissolve 8.70 g of sodium 1-pentane sul-

fonate and 8.52 g of anhydrous sodium sulfate in 980 mL ofwater, adjust the pH to 4.0 with acetic acid (100), and addwater to make 1000 mL. To 230 mL of this solution add20 mL of methanol.

Reagent: Dissolve 12.36 g of boric acid in 960 mL ofwater, add 10 mL of a solution of o-phthalaldehyde inethanol (99.5) (1 in 25), adjust the pH to 10.5 with 8 mol/Lpotassium hydroxide TS, and add water to make 1000 mL.To this solution add 1 mL of 2-mercaptoethanol.

Reaction temperature: A constant temperature of about509C.

Flow rate of the mobile phase: 0.5 mL per minute.Flow rate of the reagent: 1 mL per minute.

System suitability—System performance: Dissolve 20 mg each of Arbekacin

Sulfate, becanamycin sulfate and dibekacin sulfate in200 mL of water. When the procedure is run with 5 mL ofthis solution under the above operating conditions, be-canamycin, arbekacin and dibekacin are eluted in this order,and the resolution between the peaks, becanamycin andarbekacin is not less than 5 and arbekacin and dibekacin isnot less than 1.5, respectively.

System repeatability: When the test is repeated 6 timeswith 5 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio ofthe peak area of dibekacin to that of the internal standard isnot more than 2.0z.

(4) Related substances—Dissolve 20 mg of ArbekacinSulfate in 20 mL of water, and use this solution as the sam-ple solution. Pipet 3 mL of the sample solution, add water tomake exactly 250 mL, and use this solution as the standardsolution. Perform the test with 5 mL each of the sample solu-tion and the standard solution as directed under the LiquidChromatography according to the following conditions, anddetermine the area of each peak by the automatic integrationmethod: the total area of the peaks other than arbekacin anddibekacin obtained from the sample solution is not morethan the peak area of arbekacin from the standard solution.Operating conditions—

Detector, column, column temperature, reaction coil,reaction coil temperature, mobile phase, reagent, reactiontemperature, ‰ow rate of mobile phase, and ‰ow rate ofreagent: Proceed as directed in the operating conditions inthe Purity (3).

Time span of measurement: About 1.5 times as long as theretention time of arbekacin.System suitability—

System performance: Dissolve 10 mg each of ArbekacinSulfate and dibekacin sulfate in 200 mL of water. When theprocedure is run with 5 mL of this solution under the aboveoperating conditions, arbekacin and dibekacin are eluted inthis order with the resolution between these peaks being notless than 1.5.

System repeatability: When the test is repeated 6 times

with 5 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peakarea of arbekacin is not more than 5.0z.

Loss on drying Not more than 5.0z (0.5 g, reduced pres-sure not exceeding 0.67 kPa, 609C, 3 hours).



for test organisms [5] under (1) Agar media for seed andbase layer, having pH 7.8 – 8.0 after sterilization.

(3) Standard solutions—Weigh accurately an amount ofArbekacin Sulfate Reference Standard, previously dried,equivalent to about 20 mg (potency), dissolve in dilutedphosphate buŠer solution, pH 6.0 (1 in 2) to make exactly50 mL, and use this solution as the standard stock solution.Keep the standard stock solution at 5 to 159C and use within30 days. Take exactly a suitable amount of the standardstock solution before use, add 0.1 mol/L phosphate buŠersolution, pH 8.0 to make solutions so that each mL contains20 mg (potency) and 5 mg (potency), and use these solutionsas the high concentration standard solution and the low con-centration standard solution, respectively.

(4) Sample solutions—Weigh accurately an amount ofArbekacin Sulfate, equivalent to about 20 mg (potency), anddissolve in water to make exactly 50 mL. Take exactly a suit-able amount of this solution, add 0.1 mol/L phosphatebuŠer solution, pH 8.0 to make solutions so that each mLcontains 20 mg (potency) and 5 mg (potency), and use thesesolutions as the high concentration sample solution and thelow concentration sample solution, respectively.


Astromicin Sulfate硫酸アストロマイシン


Astromicin Sulfate contains not less than 606 mg(potency) per mg, calculated on the anhydrous basis.The potency of Astromicin Sulfate is expressed asmass (potency) of astromicin (C17H35N5O6: 405.49).

Description Astromicin Sulfate occurs as a white to lightyellowish white, powder or masses.

It is very soluble in water, sparingly soluble in ethyleneglycol, and practically insoluble in methanol and in ethanol(99.5).

It is hygroscopic.

Identiˆcation (1) Dissolve 10 mg each of Astromicin Sul-fate and Astromicin Sulfate Reference Standard in 10 mL ofwater. To 5 mL each of these solutions add water to make


100 mL, and use these solutions as the sample solution andthe standard solution. Perform the test with these solutionsas directed under the Liquid Chromatography according tothe following conditions: the retention time of astromicinobtained from the sample solution is the same with thatfrom the standard solution.Operating conditions—

Detector, column, column temperature, reaction coil,temperature of reaction coil, mobile phase, reaction reagent,reaction temperature, ‰ow rate of mobile phase, and ‰owrate of reaction reagent: Proceed as directed in the operatingconditions in the Purity (3).

(2) To 2 mL of a solution of Astromicin Sulfate (1 in100) add 2 to 3 drops of barium chloride TS: a whiteprecipitate is formed, and it does not dissolve by addition ofdilute nitric acid.



pH The pH of a solution obtained by dissolving 1.0 g ofAstromicin Sulfate in 10 mL of water is between 4.5 and 6.5.

Purity (1) Clarity and color of solution—Dissolve 1.0 gof Astromicin Sulfate in 10 mL of water: the solution is clearand colorless to pale yellow.

(2) Heavy metals—Proceed with 1.0 g of AstromicinSulfate according to Method 1, and perform the test.Prepare the control solution with 2.0 mL of Standard LeadSolution (not more than 20 ppm).

(3) Related substances—Dissolve 0.10 g of AstromicinSulfate in 100 mL of water, and use this solution as the sam-ple solution. Pipet 2 mL of the sample solution, add water tomake exactly 100 mL, and use this solution as the standardsolution. Perform the test with 10 mL each of the sample so-lution and the standard solution as directed under the LiquidChromatography according to the following conditions, anddetermine each peak area by the automatic integrationmethod: the peak areas of the related substance III, havingthe relative retention time of about 0.1, and the related sub-stance I, having the relative retention time of about 1.2 withrespect to the peak of astromicin, from the sample solutionare not more than the peak area of astromicin from thestandard solution, the peak area of the related substance II,having the related retention time of about 0.8, is not morethan 2.0 times the peak area of astromicin from the standardsolution, and the total area of the peaks other than astromi-cin from the sample solution is not more than 3.5 times thepeak area of astromicin from the standard solution.Operating conditions—

Detector: A ‰uorophotometer (excitation wavelength:340 nm; detection wavelength: 430 nm).



Reaction coil: A stainless steel tube 0.25 mm in inside

diameter and 150 cm in length.Temperature of reaction coil: 509CMobile phase: To 800 mL of a solution of anhydrous sodi-

um sulfate (71 in 2000) add 25 mL of a solution of sodium 1-heptanesulfonate (1 in 1000) and 1 mL of acetic acid (100),and add water to make 1000 mL.

Reaction reagent: Dissolve 11.2 g of potassium hydroxide,0.458 g of polyoxyethylene (23) lauryl ether, 0.300 g of o-phthalaldehyde and 1 mL of 2-mercaptoethanol in 400 mLof a solution of boric acid (31 in 1000), and add water tomake 500 mL.

Reaction temperature: 509CFlow rate of mobile phase: 0.7 mL per minuteFlow rate of reaction reagent: 0.2 mL per minuteTime span of measurement: About 2 times as long as the

retention time of astromicin.System suitability—

Test for required detectability: To 5 mL of the samplesolution add water to make 100 mL, and use this solution asthe solution for system suitability test. Pipet 2 mL of thesolution for system suitability test, and add water to makeexactly 100 mL. Conˆrm that the peak area of astromicinobtained from 10 mL of this solution is equivalent to 1.5 to2.5z of that from 10 mL of the solution for system suitabil-ity test.

System performance: To 100 mL of water add 5 mL ofthe sample solution and 2 mL of a solution of L-valine (1 in5000). When the procedure is run with 10 mL of this solutionunder the above operating conditions, L-valine and astromi-cin are eluted in this order with the resolution between thesepeaks being not less than 1.5, and when the procedure is runwith 10 mL of the solution for system suitability test underthe above operating conditions, the symmetry coe‹cient ofthe peak of astromicin is not more than 2.0.

System repeatability: When the test is repeated 6 timeswith 10 mL of the solution for system suitability test underthe above operating conditions, the relative standard devia-tion of the peak area of astromicin is not more than 2.0z.

Water Not more than 8.0z (0.2 g, volumetric titration,back titration). Use a mixture of methanol for water deter-mination and ethylene glycol for water determination (1:1)instead of methanol for water determination.




(3) Standard solutions—Weigh accurately an amount ofAstromicin Sulfate Reference Standard, equivalent to about25 mg (potency), dissolve in diluted hydrochloric acid (1 in1000) to make exactly 25 mL, and use this solution as thestandard stock solution. Keep the standard stock solution at5 – 159C, and use within 30 days. Take exactly a suitableamount of the standard stock solution before use, add0.1 mol/L phosphate buŠer solution for antibiotics, pH 8.0


to make solutions so that each mL contains 4 mg (potency)and 1 mg (potency), and use these solutions as the high con-centration standard solution and the low concentration stan-dard solution, respectively.

(4) Sample solutions—Weigh accurately an amount ofAstromicin Sulfate, equivalent to about 25 mg (potency),and dissolve in 0.1 mol/L phosphate buŠer solution forantibiotics, pH 8.0 to make exactly 25 mL. Take exactly asuitable amount of this solution, add 0.1 mol/L phosphatebuŠer solution for antibiotics, pH 8.0 to make solutions sothat each mL contains 4 mg (potency) and 1 mg (potency),and use these solutions as the high concentration samplesolution and the low concentration sample solution, respec-tively.


Bacampicillin Hydrochloride塩酸バカンピシリン


Bacampicillin Hydrochloride is a hydrochloride ofampicilline ethoxycarbonyloxyethyl ester.

Bacampicillin Hydrochloride contains not less than626 mg (potency) per mg, calculated on the anhydrousbasis. The potency of Bacampicillin Hydrochlorideis expressed as mass (potency) of ampicillin(C16H19N3O4S: 349.40).



the anhydrous basis, ethanol (95), 25 mL, 100 mm).

Change the Purity to read:

Purity (1) Heavy metals—Proceed with 1.0 g of Bacam-picillin Hydrochloride according to Method 2, and performthe test. Prepare the control solution with 2.0 mL of Stan-dard Lead Solution (not more than 20 ppm).

(2) Arsenic—Prepare the test solution with 1.0 g ofBacampicillin Hydrochloride according to Method 3, andperform the test (not more than 2 ppm).

(3) Free ampicillin—Weigh accurately about 0.1 g ofBacampicillin Hydrochloride, transfer into a 100-mLseparater, add exactly 15 mL of ice-cold water to dissolve,add and mix with exactly 10 mL of ice-cold 0.05 mol/Lphosphate buŠer solution, pH 7.0, then add 25 mL of ice-cold chloroform, shake, and abandon the chloroform layer.Repeat the procedure twice with two 25-mL portions of ice-cold chloroform. Centrifuge the water layer, ˆlter the super-natant, and use the ˆltrate as the sample solution. Separate-ly, weigh accurately an amount of Ampicillin ReferenceStandard, equivalent to about 20 mg, and dissolve in waterto make exactly 100 mL. Pipet 5 mL of this solution, add10 mL of 0.05 mol/L phosphate buŠer solution, pH 7.0 and

water to make exactly 25 mL, and use this solution as thestandard solution. To exactly 10 mL each of the samplesolution and the standard solution add exactly 2 mL ofsodium hydroxide TS, allow to stand for exactly 15 minutes,add exactly 2 mL of 1 mol/L hydrochloric acid TS, exactly10 mL of 0.3 mol/L potassium hydrogen phthalate buŠersolution, pH 4.6, and exactly 10 mL of 0.005 mol/L iodineVS, allow to stand for exactly 20 minutes without exposureto light. Titrate these solutions with 0.01 mol/L sodiumthiosulfate VS until the color of the solution changes tocolorless. Separately, to exactly 10 mL each of the samplesolution and the standard solution add exactly 10 mL of0.3 mol/L potassium hydrogen phthalate buŠer solution,pH 4.6 and exactly 10 mL of 0.005 mol/L iodine VS, andperform a blank determination with the same manner. De-termine the consumed amounts (mL) of 0.005 mol/L iodineVS, VT and VS, of the sample solution and the standard solu-tion: the amount of ampicillin is not more than 1.0z.

Amount (mg) of ampicillin (C16H19N3O4S)

＝WS×VT

VS×

120

WS: Amount (mg) of Ampicillin Reference Standard


Assay Weigh accurately an amount of BacampicillinHydrochloride and Bacampicillin Hydrochloride ReferenceStandard, equivalent to about 40 mg (potency), dissolveeach in water to make exactly 100 mL, and use these solu-tions as the sample solution and the standard solution. Per-form the test with exactly 20 mL each of the sample solutionand the standard solution as directed under the Liquid Chro-matography according to the following conditions, and cal-culate the peak areas, AT and AS, of bacampicillin of thesesolutions.


＝WS×AT

AS× 1000

WS: Amount [mg (potency)] of Bacampicillin Hydrochlo-ride Reference Standard


length: 254 nm).Column: A stainless steel column 4.6 mm in inside

diameter and 15 cm in length, packed with octadecyl-silanized silica gel for liquid chromatography (5 mm in parti-cle diameter).


Mobile phase: To 500 mL of diluted 2 mol/L sodiumdihydrogen phosphate TS (1 in 100), add diluted 0.05 mol/Ldisodium hydrogen phosphate TS (2 in 5) to adjust the pH to6.8. To 500 mL of this solution add 500 mL of acetonitrile.

Flow rate: Adjust the ‰ow rate so that the retention timeof bacampicillin is about 6.5 minutes.


System suitability—System performance: When the procedure is run with

20 mL of this solution under the above operating conditions,the number of theoretical plates and the symmetry constantof the peak of bacampicillin are not less than 10,000 and notmore than 2, respectively.

System repeatability: When the test is repeated 6 timeswith 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of peak areasof bacampicillin is not more than 2.0z.

Add the following:

Bacitracinバシトラシン

[1405-87-4]

Bacitracin is a mixture of peptide substances havingantibacterial activity including bacitracin A as themain component produced by the growth of Bacillussubtilis or Bacillus licheniformis.

It contains not less than 40 Units per mg. Thepotency of Bacitracin is expressed as unit calculatedfrom the amount of bacitracin A (C66H103N17O16S:1422.69). 1 unit of Bacitracin is equivalent to 23.8 mgof bacitracin A (C66H103N17O16S).

Description Bacitracin occurs as a white to light brownpowder.

It is freely soluble in water, and slightly soluble in ethanol(99.5).

Identiˆcation (1) To 3 mL of a solution of Bacitracin (1in 100) add 3 mL of 4-dimethylaminobenzaldehyde TS,shake until red-rosy to red-purple color appears, then addseveral drops of a solution of sodium nitrite (1 in 100), andshake: a green to dark green color is produced.

(2) Dissolve 60 mg each of Bacitracin and BacitracinReference Standard in 10 mL of water, and use thesesolutions as the sample solution and the standard solution.Perform the test with these solutions as directed under theThin-layer Chromatography. Spot 1 mL each of the samplesolution and the standard solution on a plate of silica gel forthin-layer chromatograph. Develop the plate with a mixtureof 1-butanol, acetic acid (100), water, pyridine and ethanol(99.5) (30:15:10:6:5) to a distance of about 10 cm, and air-dry the plate. Spray evenly ninhydrin TS on the plate, andheat at 1109C for 5 minutes: the spots obtained from thesample solution and the standard solution show the same Rfvalue.

Purity (1) Heavy metals—Proceed with 1.0 g of Bacitra-cin according to Method 2, and perform the test. Prepare thecontrol solution with 2.0 mL of Standard Lead Solution (notmore than 20 ppm).

(2) Dissolve 0.15 g of Bacitracin in 0.05 mol/L sulfuricacid TS to make 100 mL. To 2 mL of this solution add

0.05 mol/L sulfuric acid TS to make 10 mL, and determinethe absorbances of this solution, AT and AF, at 252 nm and290 nm as directed under the Ultraviolet-visible Spec-trophotometry: AF/AT is not more than 0.20.




(1) Test organism—Micrococcus luteus ATCC 10240.(2) Culture medium—Use the medium iii in 3) Medium

for other organisms under (1) Agar media for seed and baselayer.

(3) Standard solutions—Weigh accurately an amount ofBacitracin Reference Standard, equivalent to about 400units, dissolve in phosphate buŠer solution, pH 6.0 to makeexactly 20 mL, and use this solution as the standard stocksolution. Keep the standard stock solution at not exceeding109C and use within 2 days. Take exactly a suitable amountof the standard stock solution before use, add phosphatebuŠer solution, pH 6.0 to make solutions so that each mLcontains 2 units and 0.5 units, and use these solutions as thehigh concentration standard solution and the low concentra-tion standard solution, respectively.

(4) Sample solutions—Weigh accurately an amount ofBacitracin, equivalent about 400 units, dissolve in phosphatebuŠer solution, pH 6.0 to make exactly 20 mL. Take exactlya suitable amount of this solution, add phosphate buŠersolution, pH 6.0 to make solutions so that each mL contains2 units and 0.5 units, and use these solutions as the high con-centration sample solution and the low concentration samplesolution, respectively.

Containers and storage Containers—Tight containers.Storage—In a cold place.

Bekanamycin Sulfate硫酸ベカナマイシン


Bekanamycin Sulfate contains not less than 680 mg(potency) per mg, calculated on the dried basis. Thepotency of Bekanamycin Sulfate is expressed as mass(potency) of bekanamycin (C18H37N5O10: 483.51).

Description Bekanamycin Sulfate occurs as a white pow-der.


Identiˆcation (1) Dissolve 20 mg of Bekanamycin Sulfatein 2 mL of 1/15 mol/L phosphate buŠer solution, pH 5.6,add 1 mL of ninhydrin TS, and boil: a blue-purple color


develops.(2) Dissolve 30 mg each of Bekanamycin Sulfate and

Bekanamycin Sulfate Reference Standard in 5 mL of water,and use these solutions as the sample solution and thestandard solution. Perform the test with these solutions asdirected under the Thin-layer Chromatography. Spot 5 mLeach of the sample solution and the standard solution on aplate of silica gel for thin-layer chromatography. Developthe plate with a solution of potassium dihydrogen phosphate(3 in 40) to a distance of about 10 cm, and air-dry the plate.Spray evenly 0.2z ninhydrin-water saturated 1-butanol TS,and heat at 1009C for 10 minutes: the principal spots ob-tained from the sample solution and the standard solutionshow a purple-brown color and the same Rf value.

(3) To a solution of Bekanamycin Sulfate (1 in 5) add 1drop of barium chloride TS: a white turbidity is produced.

Optical rotation [a]20D : ＋102 –＋1169(after drying, 0.25 g,


pH The pH of a solution obtained by dissolving 0.50 g ofBekanamycin Sulfate in 10 mL of water is between 6.0 and8.5.

Purity (1) Clarity and color of solution—Dissolve 0.5 gof Bekanamycin Sulfate in 5 mL of water: the solution isclear and colorless.

(2) Heavy metals—Proceed with 1.0 g of BekanamycinSulfate according to Method 4, and perform the test. Pre-pare the control solution with 3.0 mL of Standard LeadSolution (not more than 30 ppm).

(3) Arsenic—Prepare the test solution with 2.0 g ofBekanamycin Sulfate according to Method 1, and performthe test (not more than 1 ppm).

(4) Related substances—Dissolve 60 mg of BekanamycinSulfate in 10 mL of water, and use this solution as the sam-ple solution. Pipet 3 mL of the sample solution, add water tomake exactly 100 mL, and use this solution as the standardsolution. Perform the test with these solutions as directedunder the Thin-layer Chromatography. Spot 5 mL each ofthe sample solution and the standard solution on a plate ofsilica gel for thin-layer chromatography. Develop the platewith a solution of potassium dihydrogen phosphate (3 in 40)to a distance of about 10 cm, and air-dry the plate. Sprayevenly 0.2z ninhydrin-water saturated 1-butanol TS on theplate, and heat at 1009C for 10 minutes: the spot other thanthe principal spot obtained from the sample solution is notmore intense than the spot from the standard solution.





for test organism [5] under (1) Agar media for seed and baselayer having pH 7.8 to 8.0 after sterilization.

(3) Standard solutions—Weigh accurately an amount ofBekanamycin Sulfate Reference Standard, previously dried,equivalent to about 20 mg (potency), dissolve in dilutedphosphate buŠer solution, pH 6.0 (1 in 2) to make exactly50 mL, and use this solution as the standard stock solution.Keep the standard stock solution at 5 to 159C and use within30 days. Take exactly a suitable amount of the standardstock solution before use, add 0.1 mol/L phosphate buŠersolution, pH 8.0 to make solutions so that each mL contains10 mg (potency) and 2.5 mg (potency), and use these solutionsas the high concentration standard solution and the low con-centration standard solution, respectively.

(4) Sample solutions—Weigh accurately an amount ofBekanamycin Sulfate, equivalent to about 20 mg (potency),and dissolve in water to make exactly 50 mL. Take exactly asuitable amount of this solution, add 0.1 mol/L phosphatebuŠer solution, pH 8.0 to make solutions so that each mLcontains 10 mg (potency) and 2.5 mg (potency), and use thesesolutions as the high concentration sample solution and thelow concentration sample solution, respectively.


Benzbromaroneベンズブロマロン


Benzbromarone, when dried, contains not less than98.5z and not more than 101.0z of C17H12Br2O3.

Change the Description to read:

Description Benzbromarone occurs as a white to light yel-low, crystalline powder.

It is very soluble in N,N-dimethylformamide, freely solu-ble in acetone, sparingly soluble in ethanol (99.5), and prac-tically insoluble in water.

It dissolves in dilute sodium hydroxide TS.

Change the Identiˆcation to read:

Identiˆcation (1) Determine the absorption spectrum ofa solution of Benzbromarone in 0.01 mol/L sodiumhydroxide TS (1 in 100,000) as directed under the Ultrav-iolet-visible Spectrophotometry, and compare the spectrumwith the Reference Spectrum: both spectra exhibit similarintensities of absorption at the same wavelengths.

(2) Determine the infrared absorption spectrum of Ben-zbromarone, previously dried, as directed in the potassiumbromide disk method under the Infrared Spectrophotomet-ry, and compare the spectrum with the Reference Spectrum:both spectra exhibit similar intensities of absorption at thesame wave numbers.



Purity (1) Sulfate—Dissolve 1.0 g of Benzbromarone in40 mL of acetone, and add 1 mL of dilute hydrochloric acidand water to make 50 mL. Perform the test using this solu-tion as the test solution. Prepare the control solution as fol-lows: to 0.40 mL of 0.005 mol/L sulfuric acid VS add 40 mLof acetone, 1 mL of dilute hydrochloric acid and water tomake 50 mL (not more than 0.019z).

(2) Soluble halides—Dissolve 0.5 g of Benzbromarone in40 mL of acetone, and add 6 mL of dilute nitric acid andwater to make 50 mL. Proceed with this solution as directedunder the Chloride Limit Test. Prepare the control solutionas follows: to 0.25 mL of 0.01 mol/L hydrochloric acid VSadd 40 mL of acetone, 6 mL of dilute nitric acid and waterto make 50 mL.

(3) Heavy metals—Proceed with 2.0 g of Benzbroma-rone according to Method 2, and perform the test. Preparethe control solution with 2.0 mL of Standard Lead Solution(not more than 10 ppm).

(4) Iron—Prepare the test solution with 1.0 g of Ben-zbromarone according to Method 3, and perform the testaccording to Method A. Prepare the control solution with2.0 mL of Standard Iron Solution (not more than 20 ppm).

(5) Related substances—Dissolve 0.10 g of Benzbroma-rone in 10 mL of acetone, and use this solution as the samplesolution. Pipet 1 mL of the sample solution, add acetone tomake exactly 100 mL, and use this solution as the standardsolution. Perform the test with these solutions as directedunder the Thin-layer Chromatography. Spot 10 mL each ofthe sample solution and the standard solution on a plate ofsilica gel with ‰uorescent indicator for thin-layer chro-matography. Develop the plate with a mixture of cyclo-hexane, 4-methyl-2-pentanone, ethanol (99.5) and aceticacid (100) (100:20:2:1) to a distance of about 15 cm, and air-dry the plate. Examine under ultraviolet light (main wave-length: 254 nm): the spots other than the principal spot fromthe sample solution are not more intense than the spot fromthe standard solution.

Add the following:

Benzylpenicillin Benzathineベンジルペニシリンベンザチン

(C16H18N2O4S)2.C16H20N2.4H2O: 981.18(2S,5R,6R )-3,3-Dimethyl-7-oxo-6-phenylacetylamino-4-thia-1-azabicyclo[3.2.0]heptane-2-carboxylic acidhemi(N,N?-dibenzylethylenediamine) dihydrate[41372-02-5]

Benzylpenicillin Benzathine contains not less than1152 units per mg, calculated on the anhydrous basis.The potency of Benzylpenicillin Benzathine is ex-pressed as unit calculated from the amount of benzyl-penicillin sodium (C16H17N2NaO4S: 356.37). 1 unit ofBenzylpenicillin Benzathine is equivalent to 0.6 mg ofbenzylpenicillin sodium (C16H17N2NaO4S).

Description Benzylpenicillin Benzathine occurs as a whitecrystalline powder.

It is slightly soluble in methanol and in ethanol (99.5), andpractically insoluble in water.

Identiˆcation (1) Determine the absorption spectrum ofa solution of Benzylpenicillin Benzathine in methanol (1 in2000) as directed under the Ultraviolet-visible Spectrophoto-metry, and compare the spectrum with the Reference Spec-trum: both spectra exhibit similar intensities of absorption atthe same wavelengths.

(2) Determine the infrared absorption spectrum ofBenzylpenicillin Benzathine as directed in the potassiumbromide disk method under the Infrared Spectrophotomet-ry, and compare the spectrum with the Reference Spectrum:both spectra exhibit similar intensities of absorption at thesame wave numbers.


the anhydrous basis, methanol, 20 mL, 100 mm).

Purity (1) Heavy metals—Proceed with 1.0 g of Benzyl-penicillin Benzathine according to Method 2, and performthe test. Prepare the control solution with 2.0 mL of Stan-dard Lead Solution (not more than 20 ppm).

(2) Arsenic—Prepare the test solution with 1.0 g ofBenzylpenicillin Benzathine according to Method 3, andperform the test (not more than 2 ppm).

(3) Related substances—Dissolve 70 mg of Benzyl-penicillin Benzathine in 25 mL of methanol, add a solutionprepared by dissolving 1.02 g of disodium hydrogen phos-phate and 6.80 g of potassium dihydrogen phosphate inwater to make 1000 mL to make 50 mL, and use this solu-tion as the sample solution. Pipet 1 mL of the sample solu-tion, add the mobile phase A to make exactly 100 mL, and


use this solution as the standard solution. Perform the testwith 20 mL each of the sample solution and the standardsolution as directed under the Liquid Chromatographyaccording to the following conditions, and determine eachpeak area by the automatic integration method: the area ofthe peak having the relative retention time of about 2.4 withrespect to benzylpenicillin obtained from the sample solu-tion is not more than 2 times the total area of the peaks ofbenzylpenicillin and benzathine obtained from the standardsolution, and the total area of the peaks other than benzyl-penicillin, benzathine and the peak having the relative reten-tion time of about 2.4 obtained from the sample solution isnot more than the total area of the peaks of benzylpenicillinand benzathine obtained from the standard solution.Operating conditions—




Mobile phase A: A mixture of water, methanol and0.25 mol/L potassium dihydrogen phosphate TS, pH 3.5(6:3:1).

Mobile phase B: A mixture of methanol, water and0.25 mol/L potassium dihydrogen phosphate TS, pH 3.5(6:3:1).

Flowing of the mobile phase: Control the gradient by mix-ing the mobile phases A and B as directed in the followingtable.

Time after injectionof sample (min)

Mobile phaseA (z)

Mobile phaseB (z)

0 – 10 75 2510 – 20 75→ 0 25→ 10020 – 55 0 100

Flow rate: 1.0 mL/minTime span of measurement: About 3 times as long as the

retention time of benzylpenicillin after the solvent peak.System suitability—

Test for required detectability: To exactly 1 mL of thestandard solution add the mobile phase A to make exactly20 mL. Conˆrm that the peak area of benzylpenicillinobtained from 20 mL of this solution is equivalent to 3.5 to6.5z of that from the standard solution.

System performance: When the procedure is run with20 mL of the sample solution under the above operating con-ditions, benzathine and benzylpenicillin are eluted in thisorder with the resolution between these peaks being not lessthan 25.

System repeatability: When the test is repeated 3 timeswith 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peakarea of benzylpenicillin is not more than 2.0z.

Water 5.0 – 8.0z (1 g, volumetric titration, direct titra-

tion).


(1) Test organism—Staphylococcus aureus ATCC 6538P

(2) Culture medium—Use the medium iii in 3) Mediumfor other organisms under (1) Agar media for seed and baselayer.

(3) Standard solutions—Weigh accurately an amount ofBenzylpenicillin Sodium Reference Standard, equivalent toabout 20,000 units, dissolve in phosphate buŠer solution,pH 6.0 to make exactly 10 mL, and use this solution as thestandard stock solution. Keep the standard stock solution atnot exceeding 59C and use within 2 days. Take exactly a suit-able amount of the standard stock solution before use, addphosphate buŠer solution, pH 6.0 to make solutions so thateach mL contains 2 units and 0.5 units, and use these solu-tions as the high concentration standard solution and thelow concentration standard solution, respectively.

(4) Sample solutions—Weigh accurately an amount ofBenzylpenicillin Benzathine, equivalent about 20,000 units,and dissolve in N,N-dimethylformamide to make exactly10 mL. Take exactly a suitable amount of this solution, addphosphate buŠer solution, pH 6.0 to make solutions so thateach mL contains 2 units and 0.5 units, and use these solu-tions as the high concentration sample solution and the lowconcentration sample solution, respectively.


Benzylpenicillin Potassiumベンジルペニシリンカリウム


Benzylpenicillin Potassium contains not less than1430 units per mg, calculated on the dried basis. Thepotency of Benzylpenicillin Potassium is expressedas mass unit of benzylpenicillin potassium(C16H17KN2O4S). One unit of Benzylpenicillin Potas-sium is equivalent to 0.57 mg of benzylpenicillin potas-sium.

Description Benzylpenicillin Potassium occurs as white,crystals or crystalline powder.

It is very soluble in water, and slightly soluble in ethanol(99.5).

Identiˆcation (1) Determine the absorption spectrum ofa solution of Benzylpenicillin Potassium (1 in 1000) asdirected under the Ultraviolet-visible Spectrophotometry,and compare the spectrum with the Reference Spectrum orthe spectrum of a solution of Benzylpenicillin PotassiumReference Standard prepared in the same manner as the


sample solution: both spectra exhibit similar intensities ofabsorption at the same wavelengths.

(2) Determine the infrared absorption spectrum of Ben-zylpenicillin Potassium as directed in the potassium bromidedisk method under the Infrared Spectrophotometry, andcompare the spectrum with the Reference Spectrum or thespectrum of Benzylpenicillin Potassium Reference Standard:both spectra exhibit similar intensities of absorption at thesame wave numbers.

(3) Benzylpenicillin Potassium responds to the Qualita-tive Test (1) for potassium salt.


the dried basis, water, 50 mL, 100 mm).

pH The pH of a solution obtained by dissolving 1.0 g ofBenzylpenicillin Potassium in 100 mL of water is between5.0 and 7.5.

Purity (1) Clarity and color of solution－A solutionobtained by dissolving 1 g of Benzylpenicillin Potassium in10 mL of water is clear, and colorless or light yellow.

(2) Heavy metals—Proceed with 2.0 g of Benzylpenicil-lin Potassium according to Method 4, and perform the test.Prepare the control solution with 2.0 mL of Standard LeadSolution (not more than 10 ppm).

(3) Arsenic—Prepare the test solution by incinerating1.0 g of Benzylpenicillin Potassium according to Method 4,and perform the test. In the incineration, use a crucible ofporcelain, and after addition of 10 mL of a solution ofmagnesium nitrate hexahydrate in ethanol (95) (1 in 10) add1 mL of hydrogen peroxide (30), then burn the ethanol (notmore than 2 ppm).

(4) Related substances—Dissolve 40 mg of Benzyl-penicillin Potassium in 20 mL of water, and use this solutionas the sample solution. Pipet 1 mL of the sample solution,add water to make exactly 100 mL, and use this solution asthe standard solution. Perform the test with 20 mL each ofthe sample solution and the standard solution as directedunder the Liquid Chromatography according to the follow-ing conditions, and determine each peak area by the auto-matic integration method: the area of the peak other thanbenzylpenicillin obtained from the sample solution is notmore than the peak area of benzylpenicillin from the stan-dard solution, and the total area of the peaks other than ben-zylpenicillin from the sample solution is not more than 3times the peak area of benzylpenicillin from the standardsolution.Operating conditions—




Mobile phase: A mixture of a solution of diammoniumhydrogen phosphate (33 in 5000) and acetonitrile (19:6),

adjusted the pH to 8.0 with phosphoric acid.Flow rate: Adjust the ‰ow rate so that the retention time

of benzylpenicillin is about 7.5 minutes.Time span of measurement: About 5 times as long as the

retention time of benzylpenicillin.System suitability—

Test for required detection: Pipet 10 mL of the standardsolution, and add water to make exactly 100 mL. Conˆrmthat the peak area of benzylpenicillin obtained from 20 mLof this solution is equivalent to 7 to 13z of that from 20 mLof the standard solution.

System performance: Dissolve 40 mg of BenzylpenicillinPotassium in 20 mL of water. Separately, dissolve 10 mg ofmethyl parahydroxybenzoate in 20 mL of acetonitrile. To1 mL of this solution add water to make 20 mL. Mix 1 mLeach of these solutions, and add water to make 100 mL.When the procedure is run with 20 mL of this solution underthe above operating conditions, benzylpenicillin and methylparahydroxybenzoate are eluted in this order with the reso-lution between these peaks being not less than 8.

System repeatability: When the test is repeated 5 timeswith 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peakarea of benzylpenicillin is not more than 2.0z.

Loss on drying Not more than 1.0z (3 g, reduced pressurenot exceeding 0.67 kPa, 609C, 3 hours).



(2) Culture medium—Use the medium iii in 3) Mediumfor other organisms under (1) Agar media for seed and baselayer.

(3) Standard solutions—Weigh accurately an amount ofBenzylpenicillin Potassium Reference Standard, equivalentto about 40,000 units, dissolve in phosphate buŠer solution,pH 6.0 to make exactly 100 mL, and use this solution as thestandard stock solution. Keep the standard stock solution atnot exceeding 59C and use within 2 days. Take exactly a suit-able amount of the standard stock solution before use, addphosphate buŠer solution, pH 6.0 to make solutions so thateach mL contains 2 units and 0.5 units, and use these solu-tions as the high concentration standard solution and thelow concentration standard solution, respectively.

(4) Sample solutions—Weigh accurately an amount ofBenzylpenicillin Potassium, equivalent to about 40,000units, and dissolve in phosphate buŠer solution, pH 6.0 tomake exactly 100 mL. Take exactly a suitable amount of thissolution, add phosphate buŠer solution, pH 6.0 to makesolutions so that each mL contains 2 units and 0.5 units, anduse these solutions as the high concentration sample solutionand the low concentration sample solution, respectively.



Betamethasone Sodium Phosphateリン酸ベタメタゾンナトリウム


Assay Weigh accurately about 20 mg each of Betametha-sone Sodium Phosphate and Betamethasone Sodium Phos-phate Reference Standard (determine its water content be-fore using in the same manner as Betamethasone SodiumPhosphate), and dissolve each in methanol to make exactly20 mL. Pipet 5 mL each of these solutions, and exactly 5 mLof the internal standard solution, then add methanol tomake 50 mL, and use these solutions as the sample solutionand the standard solution, respectively. Perform the testwith 10 mL each of the sample solution and the standard so-lution as directed under the Liquid Chromatography accord-ing to the following conditions, and calculate the ratios, QT

and QS, of the peak area of betamethasone phosphate to thatof the internal standard, respectively.

Amount (mg) of C22H28FNa2O8P＝WS×QT

QS

WS: Amount (mg) of Betamethasone Sodium PhosphateReference Standard, calculated on the anhydrousbasis

Internal standard solution—A solution of butyl parahydrox-ybenzoate in methanol (1 in 5000).Operating conditions—




Mobile phase: Dissolve 1.6 g of tetra-n-butylammoniumbromide, 3.2 g of disodium hydrogen phosphate 12-waterand 6.9 g of potassium dihydrogen phosphate in 1000 mL ofwater, and add 1500 mL of methanol.

Flow rate: Adjust the ‰ow rate so that the retention timeof betamethasone phosphate is about 5 minutes.System suitability—

System performance: When the procedure is run with10 mL of the standard solution under the above operatingconditions, betamethasone phosphate and the internal stan-dard are eluted in this order with the resolution betweenthese peaks being not less than 10.

System repeatability: When the test is repeated 6 timeswith 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratiosof the peak area of betamethasone phosphate to that of theinternal standard is not more than 1.0z.

Betamethasone Valerate吉草酸ベタメタゾン


Assay Dissolve about 10 mg each of Betamethasone Valer-ate and Betamethasone Valerate Reference Standard, previ-ously dried and accurately weighed, in methanol to makeexactly 100 mL. Pipet 10 mL each of these solutions, add10 mL each of the internal standard solution, and use thesesolutions as the sample solution and the standard solution,respectively. Perform the test with 10 mL each of the samplesolution and the standard solution as directed under theLiquid Chromatography according to the following condi-tions, and calculate the ratios, QT and QS, of the peak areaof betamethasone valerate to that of the internal standard,respectively.

Amount (mg) of C27H37FO6＝WS×QT

QS

WS: Amount (mg) of Betamethasone Valerate ReferenceStandard

Internal standard solution—A solution of isoamyl benzoatein methanol (1 in 1000).Operating conditions—




Mobile phase: A mixture of methanol and water (7:3).Flow rate: Adjust the ‰ow rate so that the retention time

of betamethasone valerate is about 10 minutes.System suitability—

System performance: When the procedure is run with10 mL of the standard solution under the above operatingconditions, betamethasone valerate and the internal stan-dard are eluted in this order with the resolution betweenthese peaks being not less than 5.

System repeatability: When the test is repeated 6 timeswith 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratiosof the peak area of betamethasone valerate to that of the in-ternal standard is not more than 1.0z.


Bisacodyl Suppositoriesビサコジル坐剤


Assay Weigh accurately not less than 20 Bisacodyl Sup-positories, make them ˆne fragments carefully, and mixuniformly. Weigh accurately a portion of the fragments,equivalent to about 10 mg of bisacodyl (C22H19NO4), add40 mL of tetrahydrofuran, warm to 409C, dissolve by shak-ing, cool, and add tetrahydrofuran to make exactly 50 mL.Pipet 5 mL of this solution, add exactly 5 mL of the internalstandard solution, and add the mobile phase to make100 mL. Cool this solution in ice for 30 minutes, centrifuge,ˆlter the supernatant liquid through a membrane ˆlter withpore size of 0.5 mm, discard the ˆrst 10 mL of the ˆltrate,and use the subsequent ˆltrate as the sample solution.Separately, weigh accurately about 10 mg of BisacodylReference Standard, previously dried at 1059C for 2 hours,and dissolve in tetrahydrofuran to make exactly 50 mL.Pipet 5 mL of this solution, proceed in the same manner asthe sample solution, and use this solution as the standardsolution. Perform the test with 20 mL each of the sample so-lution and the standard solution as directed under the LiquidChromatography according to the following conditions, andcalculate the ratios, QT and QS, of the peak area of bisacodylto that of the internal standard, respectively.

Amount (mg) of bisacodyl (C22H19NO4)＝WS×QT

QS

WS: Amount (mg) of Bisacodyl Reference Standard

Internal standard solution—A solution of ethyl parahydrox-ybenzoate in acetonitrile (3 in 100,000).Operating conditions—




Mobile phase: A mixture of 0.01 mol/L citric acid TS,acetonitrile and methanol (2:1:1).

Flow rate: Adjust the ‰ow rate so that the retention timeof bisacodyl is about 8 minutes.System suitability—

System performance: When the procedure is run with20 mL of the standard solution under the above operatingconditions, the internal standard and bisacodyl are eluted inthis order with the resolution between these peaks being notless than 2.0.

System repeatability: When the test is repeated 6 timeswith 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratiosof the peak area of bisacodyl to that of the internal standardis not more than 1.0z.

Bleomycin Hydrochloride塩酸ブレオマイシン


Bleomycin Hydrochloride contains not less than1400 mg (potency) and not more than 2000 mg(potency) per mg. The potency of BleomycinHydrochloride is expressed as mass (potency) ofbleomycin A2 (C55H84ClN17O21S3: 1451.00).

Description Bleomycin Hydrochloride occurs as a white toyellowish white powder.

It is freely soluble in water, and slightly soluble in ethanol(95).

It is hygroscopic.

Identiˆcation (1) To 4 mg of Bleomycin Hydrochlorideadd 5 mL of copper (II) sulfate TS, and dissolve in water tomake 100 mL. Determine the absorption spectrum of thissolution as directed under the Ultraviolet-visible Spec-trophotometry, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities ofabsorption at the same wavelengths.

(2) Determine the infrared absorption spectrum ofBleomycin Hydrochloride as directed in the potassiumbromide disk method under the Infrared Spectrophotomet-ry, and compare the spectrum with the Reference Spectrum:both spectra exhibit similar intensities of absorption at thesame wave numbers.

(3) A solution of Bleomycin Hydrochloride (1 in 100)responds to the Qualitative Test (2) for chloride.

pH The pH of a solution obtained by dissolving 0.10 g ofBleomycin Hydrochloride in 20 mL of water is between 4.5and 6.0.

Content ratio of the active principle Dissolve 10 mg ofBleomycin Hydrochloride in 20 mL of water, and use thissolution as the sample solution. Perform the test with 20 mLof the sample solution as directed under the Liquid Chro-matography according to the following conditions, and de-termine each peak area by the automatic integrationmethod: the peak area of bleomycin A2 (the ˆrst principalpeak) is between 55z and 70z, that of bleomycin B2 (thesecond principal peak) is between 25z and 32z, the totalpeak area of bleomycin A2 and bleomycin B2 is not less than85z, the peak area of demethylbleomycin A2 (a peak havingthe relative retention time of 1.5 – 2.5 to bleomycin A2) isnot more than 5.5z, and the total area of the rest peaks isnot more than 9.5z.Operating conditions—


Column: A stainless steel column 4.6 mm in inside di-ameter and 25 cm in length, packed with octadecylsilanizedsilica gel for liquid chromatography (7 mm in particle


diameter).Column temperature: A constant temperature of about

409C.Mobile phase stock solution: Dissolve 0.96 g of sodium

1-pentanesulfonate and 1.86 g of disodium dihydrogenethylenediamine tetraacetate dihydrate in 1000 mL of waterand 5 mL of acetic acid (100), and adjust the pH to 4.3 withammonia TS.

Mobile phase A: A mixture of the mobile phase stocksolution and methanol (9:1).

Mobile phase B: A mixture of the mobile phase stock solu-tion and methanol (3:2).

Flowing of the mobile phase: Control the gradient bymixing the mobile phases A and B as directed in the follow-ing table.


Mobile phaseA (z)

Mobile phaseB (z)

0 – 60 100→ 0 0→ 10060 – 75 0 100

Flow rate: About 1.2 mL per minute.Time span of measurement: 20 minutes after elution of

the peak of demethylbreomycin A2 after the solvent peak.System suitability—

System performance: When the procedure is run with20 mL of the sample solution under the above operating con-ditions, bleomycin A2 and bleomycin B2 are eluted in thisorder with the resolution between these peaks being not lessthan 5.

System repeatability: When the test is repeated 6 timeswith 20 mL of the sample solution under the above operatingconditions, the relative standard deviation of the peak areaof bleomycin A2 is not more than 2.0z.

Purity (1) Clarity and color of solution—A solutionobtained by dissolving 0.08 g of Bleomycin Hydrochloridein 4 mL of water is clear and colorless.

(2) Copper—Dissolve exactly 75 mg of BleomycinHydrochloride in exactly 10 mL of diluted nitric acid (1 in100), and use this solution as the sample solution. Separate-ly, to exactly 15 mL of Standard Copper Solution add dilut-ed nitric acid (1 in 100) to make exactly 100 mL, and use thissolution as the standard solution. Perform the test with thesample solution and the standard solution as directed underthe Atomic Absorption Spectrophotometry according to thefollowing conditions: the absorbance of the sample solutionis not more than that of the standard solution (not morethan 200 ppm).

Gas: Combustible gas—AcetyleneSupporting gas—Air

Lamp: Copper hollow-cathode lampWavelength: 324.8 nm

Loss on drying Not more than 5.0z (60 mg, in vacuum,phosphorus (V) oxide, 609C, 3 hours). Take the sample to betested while avoiding moisture absorption.

Assay Perform the test according to the Cylinder-platemethod as directed under the Microbial Assay for Antibiot-

ics according to the following conditions.(1) Test organism—Mycobacterium smegmatis ATCC

607(2) Agar medium for seed, base layer and transferring

the test organismGlycerin 10.0 gPeptone 10.0 gMeat extract 10.0 gSodium chloride 3.0 gAgar 15.0 gWater 1000 mL

Mix all the components, and sterilize. Adjust the pH aftersterilization to 6.9 – 7.1 with sodium hydroxide TS.

(3) Liquid media for suspending the test organismGlycerin 10.0 gPeptone 10.0 gMeat extract 10.0 gSodium chloride 3.0 gWater 1000 mL

Mix all the components, and sterilize. Adjust the pH aftersterilization to 6.9 – 7.1 with sodium hydroxide TS.

(4) Preparation of seeded agar layer—Cultivate the testorganism on the slant of the agar medium for transferringthe test organism at 279C for 40 to 48 hours, then inoculatethe test organism thus obtained in 100 mL of the liquidmedia for suspending the test organism, cultivate with shak-ing at between 259C and 279C for 5 days, and use this as thesuspension of test organism. Store the suspension of testorganism at a temperature not exceeding 59C, and usewithin 14 days. Add 0.5 mL of the suspension of test organ-ism in 100 mL of the agar medium for seed previously keptat 489C, mix thoroughly, and use as the seeded agar layer.

(5) Preparation of cylinder-agar plate—Proceed asdirected in the Preparation of cylinder-agar plate under theMicrobial Assay for Antibiotics, dispensing 5.0 mL of agarmedium for base layer and 8.0 mL of the agar medium forseed into the Petri dish.

(6) Standard solutions—Weigh accurately an amount ofBleomycin A2 Hydrochloride Reference Standard, previous-ly dried under reduced pressure not exceeding 0.67 kPa at anordinary temperature for 3 hours, equivalent to about 15 mg(potency), dissolve in 0.1 mol/L phosphate buŠer solution,pH 6.8 to make exactly 100 mL, and use this solution as thestandard stock solution. Keep the standard stock solution at59C or below, and use within 30 days. Take exactly a suita-ble amount of the standard stock solution before use, add0.1 mol/L phosphate buŠer solution, pH 6.8 to make solu-tions so that each mL contains 30 mg (potency) and 15 mg(potency), and use these solutions as the high concentrationstandard solution and the low concentration standard solu-tion, respectively.

(7) Sample solutions—Weigh accurately an amount ofBleomycin Hydrochloride, equivalent to about 15 mg(potency), and dissolve in 0.1 mol/L phosphate buŠer solu-tion, pH 6.8 to make exactly 100 mL. Take exactly a suitableamount of this solution, add 0.1 mol/L phosphate buŠersolution, pH 6.8 to make solutions so that each mL contains30 mg (potency) and 15 mg (potency), and use these solutions


as the high concentration sample solution and the low con-centration sample solution, respectively.


Bleomycin Sulfate硫酸ブレオマイシン


Bleomycin Sulfate contains not less than 1400 mg(potency) and not more than 2000 mg (potency) permg. The potency of Bleomycin Sulfate is expressed asmass (potency) of bleomycin A2 (C55H84ClN17O21S3:1451.00).

Description Bleomycin Sulfate occurs as a white to yellow-ish white powder.

It is freely soluble in water, and slightly soluble in ethanol(95).

It is hygroscopic.

Identiˆcation (1) To 4 mg of Bleomycin Sulfate add 5 mLof copper (II) sulfate TS, and dissolve in water to make100 mL. Determine the absorption spectrum of this solutionas directed under the Ultraviolet-visible Spectrophotometry,and compare the spectrum with the Reference Spectrum:both spectra exhibit similar intensities of absorption at thesame wavelengths.

(2) Determine the infrared absorption spectrum ofBleomycin Sulfate as directed in the potassium bromide diskmethod under the Infrared Spectrophotometry, and com-pare the spectrum with the Reference Spectrum: both spec-tra exhibit similar intensities of absorption at the same wavenumbers.

(3) Bleomycin Sulfate responds to the Qualitative Tests(1) and (2) for sulfate.

pH The pH of a solution obtained by dissolving 10 mg ofBleomycin Sulfate in 20 mL of water is between 4.5 and 6.0.

Content ratio of the active principle Dissolve 10 mg ofBleomycin Sulfate in 20 mL of water, and use this solutionas the sample solution. Perform the test with 20 mL of thesample solution as directed under the Liquid Chro-matography according to the following conditions, and de-termine each peak area by the automatic integrationmethod: the peak area of bleomycin A2 (the ˆrst principalpeak) is between 55z and 70z, that of bleomycin B2 (thesecond principal peak) is between 25z and 32z, the totalpeak area of bleomycin A2 and bleomycin B2 is not less than85z, the peak area of demethylbleomycin A2 (a peak havingthe relative retention time of 1.5 – 2.5 against bleomycin A2)is not more than 5.5z, and the total area of the rest peaks isnot more than 9.5z.Operating conditions—

Detector: An ultraviolet absorption photometer (wave-


ameter and 25 cm in length, packed with octadecylsilanizedsilica gel for liquid chromatography (7 mm in particle di-ameter).


Mobile phase stock solution: Dissolve 0.96 g of sodium 1-pentanesulfonate and 1.86 g of disodium dihydrogenethylenediamine tetraacetate dihydrate in 1000 mL of waterand 5 mL of acetic acid (100), and adjust the pH to 4.3 withammonia TS.

Mobile phase A: A mixture of the mobile phase stocksolution and methanol (9:1).

Mobile phase B: A mixture of the mobile phase stock solu-tion and methanol (3:2).



Mobile phaseA (z)

Mobile phaseB (z)

0 – 60 100→ 0 0→ 10060 – 75 0 100

Flow rate: About 1.2 mL/minTime span of measurement: Twenty minutes after elution

of the peak of demethylbleomycin A2 after the solvent peak.System suitability—

System performance: When the procedure is run with20 mL of the sample solution under the above operating con-ditions, bleomycin A2 and bleomycin B2 are eluted in thisorder with the resolution between these peaks being not lessthan 5.

System repeatability: When the test is repeated 6 timeswith 20 mL of the sample solution under the above operatingconditions, the relative standard deviation of the peak areaof bleomycin A2 is not more than 2.0z.

Purity (1) Clarity and color of solution—A solutionobtained by dissolving 0.08 g of Bleomycin Sulfate in 4 mLof water is clear and colorless.

(2) Copper—Dissolve exactly 75 mg of Bleomycin Sul-fate in 10 mL of diluted nitric acid (1 in 100), and use thissolution as the sample solution. Separately, to exactly 15 mLof Standard Copper Solution add diluted nitric acid (1 in100) to make exactly 100 mL, and use this solution as thestandard solution. Perform the test with the sample solutionand the standard solution as directed under the Atomic Ab-sorption Spectrophotometry according to the following con-ditions: the absorbance of the sample solution is not morethan that of the standard solution (not more than 200 ppm).


Lamp: Copper hollow-cathode lampWavelength: 324.8 nm

Loss on drying Not more than 3.0z (60 mg, in vacuum,phosphorus (V) oxide, 609C, 3 hours). Take the sample to be


tested while avoiding moisture absorption.


(1) Test organism—Mycrobacterium smegmatis ATCC607

(2) Agar medium for seed, base layer and transferringthe test organism

Glycerin 10.0 gPeptone 10.0 gMeat extract 10.0 gSodium chloride 3.0 gAgar 15.0 gWater 1000 mL

Mix all the components and sterilize. Adjust the pH aftersterilization to 6.9 – 7.1 with sodium hydroxide TS.

(3) Liquid media for suspending the test organismGlycerin 10.0 gPeptone 10.0 gMeat extract 10.0 gSodium chloride 3.0 gWater 1000 mL

Mix all the components and sterilize. Adjust the pH aftersterilization to 6.9 – 7.1 with sodium hydroxide TS.

(4) Preparation of seeded agar layer－Cultivate the testorganism on the slant of the agar medium for transferringthe test organism at 279C for 40 to 48 hours, then inoculatethe test organism thus obtained in 100 mL of the liquidmedia for suspending the test organism, cultivate with shak-ing at between 259C and 279C for 5 days, and use this as thesuspension of test organism. Store the suspension of testorganism at a temperature not exceeding 59C, and usewithin 14 days. Add 0.5 mL of the suspension of test organ-ism in 100 mL of the agar medium for seed previously keptat 489C, mix thoroughly, and use as the seeded agar layer.

(5) Preparation of cylinder-agar plate—Proceed asdirected in 7. Preparation of cylinder-agar plate under theMicrobial Assay for Antibiotics, dispensing 5.0 mL of agarmedium for base layer and 8.0 mL of the agar medium forseed into the Petri dish.

(6) Standard solutions—Weigh accurately an amount ofBleomycin A2 Hydrochloride Reference Standard, previous-ly dried under reduced pressure not exceeding 0.67 kPa at anordinary temperature for 3 hours, equivalent to about 15 mg(potency), dissolve in 0.1 mol/L phosphate buŠer solution,pH 6.8 to make exactly 100 mL, and use this solution as thestandard stock solution. Keep the standard stock solution at59C or below, and use within 30 days. Take exactly a suita-ble amount of the standard stock solution before use, add0.1 mol/L phosphate buŠer solution, pH 6.8 to make solu-tions so that each mL contains 30 mg (potency) and 15 mg(potency), and use these solutions as the high concentrationstandard solution and the low concentration standard solu-tion, respectively.

(7) Sample solutions—Weigh accurately an amount ofBleomycin Sulfate, equivalent to about 15 mg (potency), dis-solve in 0.1 mol/L phosphate buŠer solution, pH 6.8 to

make exactly 100 mL. Take exactly a suitable amount of thissolution, add 0.1 mol/L phosphate buŠer solution, pH 6.8to make solutions so that each mL contains 30 mg (potency)and 15 mg (potency), and use these solutions as the high con-centration sample solution and the low concentration samplesolution, respectively.


Calcium Chloride Injection塩化カルシウム注射液


Calcium Chloride Injection is an aqueous solutionfor injection. It contains not less than 95.0z and notmore than 105.0z of the labeled amount of calciumchloride (CaCl2: 110.98).

The concentration of Calcium Chloride Injection isexpressed as the quantity of calcium chloride (CaCl2).


Description Calcium Chloride Injection is a clear, colorlessliquid.


pH 4.5 – 7.5

Bacterial endotoxins Less than 0.30 EU/mg.

Change the Containers and storage to read:

Containers and storage Containers—Hermetic containers.Plastic containers for aqueous injections may be used.

Calcium Polystyrene Sulfonateポリスチレンスルホン酸カルシウム

Change the Purity (4) to read:

Purity(4) Styrene—To 10.0 g of Calcium Polystyrene Sul-

fonate add 10 mL of acetone, shake for 30 minutes, cen-trifuge, and use the supernatant liquid as the sample solu-tion. Separately, dissolve 10 mg of styrene in acetone tomake exactly 100 mL. Pipet 1 mL of this solution, dilutewith acetone to make exactly 100 mL, and use this solutionas the standard solution. Perform the test with exactly 5 mLeach of the sample solution and the standard solution asdirected under the Gas Chromatography according to thefollowing conditions. Determine the peak heights, HT andHS, of styrene in each solution: HT is not larger than HS.Operating conditions—

Detector: A hydrogen ‰ame-ionization detector.Column: A stainless steel column 3 mm in inside diameter


and 2 m in length, having polyethylene glycol 20 M coated atthe ratio of 15z on siliceous earth for gas chromatography(150 to 180 mm in particle diameter).


Carrier gas: NitrogenFlow rate: Adjust the ‰ow rate so that the retention time

of styrene is about 9 minutes.System suitability—

System performance: Mix 10 mg of styrene with 1000 mLof acetone. When the procedure is run with 5 mL of this solu-tion under the above operating conditions, the number oftheoretical plates and the symmetry coe‹cient of the peak ofstyrene are not less than 800 and 0.8 to 1.2, respectively.

System repeatability: When the test is repeated 6 timeswith 5 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peakheights of styrene is not more than 5z.

d-Camphord-カンフル


Purity(2) Chlorinated compounds—Mix 0.20 g of ˆnely pow-

dered d-Camphor with 0.4 g of sodium peroxide in a driedporcelain crucible. Heat the crucible gently by the open‰ame until the incineration is complete. Dissolve the residuein 20 mL of warm water, acidify with 12 mL of dilute nitricacid, and ˆlter the solution into a Nessler tube. Wash theˆlter paper with three 5-mL portions of hot water, addingthe washings to the ˆltrate. After cooling, add water to make50 mL, then add 1 mL of silver nitrate TS, mix well, andallow to stand for 5 minutes: the turbidity of the solutiondoes not exceed that of the following control solution.

Control solution: Prepare in the same manner as de-scribed above, using 0.20 mL of 0.01 mol/L hydrochloricacid VS.

dl-Camphordl-カンフル


Purity(2) Chlorinated compounds—Mix 0.20 g of ˆnely pow-

dered dl-Camphor with 0.4 g of sodium peroxide in a driedporcelain crucible. Heat the crucible gently by the open‰ame until the incineration is complete. Dissolve the residuein 20 mL of warm water, acidify with 12 mL of dilute nitricacid, and ˆlter the solution into a Nessler tube. Wash theˆlter paper with three 5-mL portions of hot water, addingthe washings to the ˆltrate. After cooling, add water to make50 mL, then add 1 mL of silver nitrate TS, mix well, and

allow to stand for 5 minutes: the turbidity of the solutiondoes not exceed that of the following control solution.

Control solution: Prepare in the same manner as de-scribed above, using 0.20 mL of 0.01 mol/L hydrochloricacid VS.

Carbazochrome Sodium Sulfonateカルバゾクロムスルホン酸ナトリウム


Purity(3) Related substances—Dissolve 50 mg of Carbazo-

chrome Sodium Sulfonate in 100 mL of water, and use thissolution as the sample solution. Pipet 2 mL of the samplesolution, add water to make exactly 200 mL, and use thissolution as the standard solution. Perform the test with ex-actly 10 mL each of the sample solution and the standard so-lution as directed under the Liquid Chromatography accord-ing to the following conditions. Determine each peak area ofthese solutions by the automatic integration method: thetotal area of the peaks other than the peak of car-bazochrome sulfonate from the sample solution is not largerthan the peak area of carbazochrome sulfonate from thestandard solution.Operating conditions—




Mobile phase: Dissolve 1.2 g of ammonium dihydrogenphosphate in 1000 mL of water, and ˆlter through a mem-brane ˆlter (0.4 mm in pore size) if necessary. To 925 mL ofthis solution add 75 mL of ethanol (95), shake, and adjustthe pH to 3 with phosphoric acid.

Flow rate: Adjust the ‰ow rate so that the retention timeof carbazochrome sulfonate is about 7 minutes.

Time span of measurement: About 3 times as long as theretention time of carbazochrome sulfonate after the solventpeak.System suitability—

Test for required detection: To exactly 2 mL of the stan-dard solution add the mobile phase to make exactly 20 mL.Conˆrm that the peak area of carbazochrome sulfonateobtained from 10 mL of this solution is equivalent to 7 to13z of that of carbazochrome sulfonate obtained from10 mL of the standard solution.

System performance: Dissolve 10 mg each of Car-bazochrome Sodium Sulfonate and carbazochrome in100 mL of water by warming. When the procedure is runwith 10 mL of this solution under the above operating condi-tions, carbazochrome sulfonate and carbazochrome are elut-


ed in this order with the resolution between these peaksbeing not less than 3.

System repeatability: When the test is repeated 6 timeswith 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peakarea of carbazochrome sulfonate is not more than 2.0z.

Carbidopaカルビドパ


Purity(2) Related substances—Dissolve 50 mg of Carbidopa in

70 mL of the mobile phase, by warming and using ultrasoni-cation, if necessary. After cooling, add the mobile phase tomake 100 mL, and use this solution as the sample solution.Pipet 1 mL of the sample solution, add the mobile phase tomake exactly 100 mL, and use this solution as the standardsolution. Perform the test with exactly 20 mL each of thesample solution and the standard solution as directed underthe Liquid Chromatography according to the following con-ditions. Determine each peak area from both solutions bythe automatic integration method: the total area of all peaksother than the peak of carbidopa from the sample solution isnot larger than the peak area of carbidopa from the standardsolution.Operating conditions—


Time span of measurement: About 3 times as long as theretention time of carbidopa.System suitability—

Test for required detection: To exactly 2 mL of the stan-dard solution add the mobile phase to make exactly 20 mL.Conˆrm that the peak area of carbidopa obtained from20 mL of this solution is equivalent to 7 to 13z of that ofcarbidopa obtained from 20 mL of the standard solution.



Assay Weigh accurately about 50 mg each of Carbidopaand Carbidopa Reference Standard (determined separatelythe loss on drying), and dissolve each in 70 mL of the mobilephase, by warming and using ultrasonication if necessary.After cooling, add the mobile phase to make exactly 100mL, and use these solutions as the sample solution and thestandard solution, respectively. Perform the test with exactly20 mL each of the sample solution and the standard solutionas directed under the Liquid Chromatography according tothe following conditions, and determine the peak areas, AT

and AS, of carbidopa in each solution.

Amount (mg) of C10H14N2O4.H2O＝WS×AT

AS× 1.080

WS: Amount (mg) of Carbidopa Reference Standard, cal-culated on the dried basis


length: 280 nm).Column: A stainless steel column 4 mm in inside diameter

and 25 cm in length, packed with octadecylsilanized silica gelfor liquid chromatography (7 mm in particle diameter).


Mobile phase: To 950 mL of 0.05 mol/L sodium dihydro-gen phosphate TS add 50 mL of ethanol (95), and adjust thepH to 2.7 with phosphoric acid.

Flow rate: Adjust the ‰ow rate so that the retention timeof carbidopa is about 6 minutes.System suitability—

System performance: Dissolve 50 mg each of Carbidopaand methyldopa in 100 mL of the mobile phase. When theprocedure is run with 20 mL of this solution under the aboveoperating conditions, methyldopa and carbidopa are elutedin this order with the resolution between these peaks beingnot less than 0.9.

System repeatability: When the test is repeated 6 timeswith 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peakarea of carbidopa is not more than 1.0z.

Carumonam Sodiumカルモナムナトリウム


Carumonam Sodium contains not less than 850 mg(potency) per mg, calculated on the anhydrous basis.The potency of Carumonam Sodium is expressed asmass (potency) of carumonam (C12H14N6O10S2:466.40).

Description Carumonam Sodium occurs as a white to yel-lowish white, crystals or crystalline powder.

It is freely soluble in water, soluble in formamide, veryslightly soluble in methanol, and practically insoluble inacetic acid (100) and in ethanol (99.5).

Identiˆcation (1) Determine the absorption spectrum ofa solution of Carumonam Sodium (3 in 100,000) as directedunder the Ultraviolet-visible Spectrophotometry, and com-pare the spectrum with the Reference Spectrum or the spec-trum of a solution of Carumonam Sodium Reference Stan-dard prepared in the same manner as the sample solution:both spectra exhibit similar intensities of absorption at thesame wavelengths.


(2) Determine the infrared absorption spectrum ofCarumonam Sodium as directed in the potassium bromidedisk method under the Infrared Spectrophotometry, andcompare the spectrum with the Reference Spectrum or thespectrum of Carumonam Sodium Reference Standard: bothspectra exhibit similar intensities of absorption at the samewave numbers.

(3) Determine the spectrum of a solution of CarumonamSodium in heavy water for nuclear magnetic resonance spec-troscopy (1 in 10) as directed under the Nuclear MagneticResonance Spectroscopy (1H), using sodium 3-trimethylsilyl-propionate-d4 for nuclear magnetic resonance spectroscopyas an internal reference compound: it exhibits a double sig-nal A at around d5.5 ppm, and a single signal B at aroundd7.0 ppm. The ratio of the integrated intensity of these sig-nals, A:B, is about 1:1.

(4) Carumonam Sodium responds to the QualitativeTest (1) for sodium salt.

Optical rotation [a]20D : ＋18.5 –＋21.09(0.1 g calculated on


pH The pH of a solution obtained by dissolving 1.0 g ofCarumonam Sodium in 10 mL of water is between 5.0 and6.5.

Purity (1) Clarity and color of solution—Dissolve 0.5 gof Carumonam Sodium in 5 mL of water: the solution isclear and colorless to pale yellow.

(2) Heavy metals—Proceed with 2.0 g of CarumonamSodium according to Method 2, and perform the test. Pre-pare the control solution with 3.0 mL of Standard LeadSolution (not more than 15 ppm).

(3) Arsenic—Prepare the test solution with 2.0 g ofCarumonam Sodium according to Method 4, and performthe test (not more than 1 ppm).

(4) Related substance 1—Weigh accurately about 0.1 gof Carumonam Sodium, and dissolve in the mobile phase tomake exactly 50 mL. Pipet 5 mL of this solution, add themobile phase to make exactly 25 mL, and use this solution asthe sample solution. Separately, weigh accurately about0.1 g of Carumonam Sodium Reference Standard, and dis-solve in the mobile phase to make exactly 50 mL. Pipet 5 mLof this solution, and add the mobile phase to make exactly25 mL. Pipet 1 mL of this solution, add the mobile phase tomake exactly 100 mL, and use this solution as the standardsolution. Perform the test with exactly 10 mL each of thesample solution and the standard solution as directed underthe Liquid Chromatography according to the following con-ditions, and determine each peak area by the automatic in-tegration method. Calculate the amount of the related sub-stances by the following equation: the amount of the relatedsubstance having the relative retention time of 0.7 to thepeak of carumonam is not more than 4.0z, and eachamount of the related substances other than the related sub-stance having the relative retention time of 0.7 to the peak ofcarumonam is not more than 1.0z.

Amount (z) of related substance ＝WS

WT×

AT

AS

WS: Amount (g) of Carumonam Sodium Reference Stan-dard

WT: Amount (g) of the sampleAS: Peak area of carumonam from the standard solutionAT: Each peak area other than carumonam from the sam-

ple solution

Operating conditions—Detector, column, column temperature, mobile phase,

and ‰ow rate: Proceed as directed in the operating condi-tions in the Assay.

Time span of measurement: About 3 times as long as theretention time of carumonam.System suitability—

Test for required detectability: Measure exactly 5 mL ofthe standard solution, and add the mobile phase to makeexactly 50 mL. Conˆrm that the peak area of carumonamobtained from 10 mL of this solution is equivalent to 7 to13z of that from 10 mL of the standard solution.

System performance: Dissolve 40 mg of Carumonam So-dium in 20 mL of the mobile phase. To 5 mL of this solutionadd 5 mL of a solution of resorcinol in the mobile phase (9 in1000) and the mobile phase to make 25 mL. When the proce-dure is run with 10 mL of this solution under the above oper-ating conditions, resorcinol and carumonam are eluted inthis order with the resolution between these peaks being notless than 2.5.

System repeatability: When the test is repeated 3 timeswith 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peakarea of carumonam is not more than 2.9z.

(5) Related substance 2—Weigh accurately about 0.1 gof Carumonam Sodium, and dissolve in the mobile phase tomake exactly 50 mL. Pipet 5 mL of this solution, add themobile phase to make exactly 25 mL, and use this solution asthe sample solution. Separately, weigh accurately about0.1 g of Carumonam Sodium Reference Standard, and dis-solve in the mobile phase to make exactly 50 mL. Pipet 5 mLof this solution, and add the mobile phase to make exactly25 mL. Pipet 1 mL of this solution, add the mobile phase tomake exactly 100 mL, and use this solution as the standardsolution. Perform the test with exactly 10 mL each of thesample solution and the standard solution as directed underthe Liquid Chromatography according to the following con-ditions, and determine each peak area by the automatic in-tegration method. Calculate the amount of the related sub-stances by the following equation: the amount of each relat-ed substance is not more than 1.0z.

Amount (z) of related substance＝WS

WT×

AT

AS

WS: Amount (g) of Carumonam Sodium Reference Stan-dard

WT: Amount (g) of the sampleAS: Peak area of carumonam from the standard solutionAT: Each area of the peaks appeared after the peak of

carumonam from the sample solution


Operating conditions—Detector, column, and column temperature: Proceed as

directed in the operating conditions in the Assay.Mobile phase: A mixture of a solution of ammonium sul-

fate (1 in 10,000), methanol and acetic acid (100) (74:25:1).Flow rate: Dissolve 0.01 g of phthalic acid in the mobile

phase to make 100 mL. Adjust the ‰ow rate so that theretention time of phthalic acid is about 6.5 minutes when theprocedure is run with 10 mL of this solution.

Time span of measurement: About 10 times as long as theretention time of carumonam.System suitability—

Test for required detectability: Measure exactly 5 mL ofthe standard solution, and add the mobile phase to makeexactly 50 mL. Conˆrm that the peak area of carumonamobtained from 10 mL of this solution is equivalent to 7 to13z of that from 10 mL of the standard solution.

System performance: Dissolve 40 mg of Carumonam So-dium in 20 mL of the mobile phase. To 5 mL of this solutionadd 5 mL of a solution of resorcinol in the mobile phase (9 in1000) and the mobile phase to make 25 mL. When the proce-dure is run with 10 mL of this solution under the above oper-ating conditions, resorcinol and carumonam are eluted inthis order with the resolution between these peaks being notless than 7.

System repeatability: When the test is repeated 3 timeswith 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peakarea of carumonam is not more than 2.9z.

(6) Total amount of related substances－The total of theamounts of the related substances obtained in the Relatedsubstance 1 and the Related substance 2 is not more than6.0z.

Water Not more than 2.0z (0.2 g, volumetric titration,direct titration; Use a mixture of formamide for water deter-mination and methanol for water determination (3:1) in-stead of methanol for water determination).

Assay Weigh accurately an amount of CarumonamSodium and Carumonam Sodium Reference Standard,equivalent to about 40 mg (potency), and dissolve each in themobile phase to make exactly 20 mL. Measure exactly 5 mLeach of these solutions, add exactly 5 mL of the internalstandard solution and the mobile phase to make 25 mL, anduse these solutions as the sample solution and the standardsolution. Perform the test with 10 mL each of the samplesolution and the standard solution as directed under the Liq-uid Chromatography according to the following conditions,and determine the ratios, QT and QS, of the peak area ofcarumonam to that of the internal standard.

Amount [mg (potency)] of carumonam (C12H14N6O10S2)

＝WS×QT

QS× 1000

WS: Amount [mg (potency)] of Carumonam SodiumReference Standard

Internal standard solution—A solution of resorcinol in themobile phase (9 in 1000).Operating conditions—




Mobile phase: A mixture of a solution of ammonium sul-fate (1 in 10,000), methanol and acetic acid (100) (97:2:1).

Flow rate: Adjust the ‰ow rate so that the retention timeof carumonam is about 10 minutes.System suitability—

System performance: When the procedure is run with10 mL of the standard solution under the above operatingconditions, the internal standard and carumonam are elutedin this order with the resolution between these peaks beingnot less than 2.5.

System repeatability: When the test is repeated 6 timeswith 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratiosof the peak area of carumonam to that of the internal stan-dard is not more than 1.0z.

Containers and storage Containers—Hermetic containers.Storage—Light-resistant.

Cefaclorセファクロル


Cefaclor contains not less than 950 mg (potency) permg, calculated on the anhydrous basis. The potency ofCefaclor is expressed as mass (potency) of cefaclor(C15H14ClN3O4S).

Description Cefaclor occurs as a white to yellowish whitecrystalline powder.

It is slightly soluble in water and in methanol, and practi-cally insoluble in ethanol (99.5) and in N,N-dimethylfor-mamide.

Identiˆcation (1) Determine the absorption spectrum ofa solution of Cefaclor (1 in 50,000) as directed under theUltraviolet-visible Spectrophotometry, and compare thespectrum with the Reference Spectrum: both spectra exhibitsimilar intensities of absorption at the same wavelengths.

(2) Determine the infrared absorption spectrum ofCefaclor as directed in the potassium bromide disk methodunder the Infrared Spectrophotometry, and compare thespectrum with the Reference Spectrum: both spectra exhibitsimilar intensities of absorption at the same wave numbers.


(3) Dissolve 0.04 g of Cefaclor in 0.5 mL of heavy waterfor nuclear magnetic resonance spectroscopy and 1 drop ofdeuterated hydrochloric acid for nuclear magnetic resonancespectroscopy, and determine the spectrum of this solution asdirected under the Nuclear Magnetic Resonance Spec-troscopy (1H), using sodium 3-trimethylsilylpropanesul-fonate for nuclear magnetic resonance spectroscopy as aninternal reference compound: it exhibits an AB type quartetsignal A at around d3.7 ppm, and a single signal or a sharpmultiple signal B at around d7.6 ppm. The ratio of theintegrated intensity of each signal, A:B, is about 2:5.

(4) Perform the test with Cefaclor as directed under theFlame Coloration Test (2): a green color appears.



Purity (1) Heavy metals—Proceed with 1.0 g of Cefacloraccording to Method 2, and perform the test. Prepare thecontrol solution with 2.0 mL of Standard Lead Solution (notmore than 20 ppm).

(2) Arsenic—Prepare the test solution by suspending1.0 g of Cefaclor in 10 mL of N,N-dimethylformamide, andperform the test (not more than 2 ppm).

(3) Related substances—Dissolve 0.05 g of Cefaclor in10 mL of sodium dihydrogen phosphate TS, pH 2.5, and usethis solution as the sample solution. Pipet 1 mL of thesample solution, add sodium dihydrogen phosphate TS, pH2.5 to make exactly 100 mL, and use this solution as thestandard solution. Perform the test with exactly 20 mL eachof the sample solution and the standard solution as directedunder the Liquid Chromatography according to the follow-ing conditions, and determine each peak area by the auto-matic integration method: the peak areas other than cefaclorfrom the sample solution are not more than 1/2 of the peakarea of cefaclor from the standard solution, and the total ofthe peak areas other than cefaclor from the sample solutionis not more than 2 times of the peak area of cefaclor fromthe standard solution. If necessary, proceed with 20 mL ofsodium dihydrogen phosphate TS, pH 2.5 in the same man-ner as above to compensate the base line.Operating conditions—




Mobile phase A: Dissolve 7.8 g of sodium dihydrogenphosphate dihydrate in 1000 mL of water, and adjust the pHto 4.0 with phosphoric acid.

Mobile phase B: To 550 mL of the mobile phase A add450 mL of acetonitrile for liquid chromatography.



Mobile phaseA (z)

Mobile phaseB (z)

0 – 30 95→ 75 5→ 2530 – 45 75→ 0 25→ 10045 – 55 0 100

Flow rate: 1.0 mL per minute.Time span of measurement: About 2.5 times as long as the

retention time of cefaclor after the solvent peak.System suitability—

Test for required detectability: Measure exactly 1 mL ofthe standard solution, and add sodium dihydrogen phos-phate TS, pH 2.5 to make exactly 20 mL. Conˆrm that thepeak area of cefaclor obtained from 20 mL of this solution isequivalent to 4 to 6z of that from 20 mL of the standardsolution.

System performance: When the procedure is run with20 mL of the standard solution under the above operatingconditions, the number of theoretical plates and the sym-metry coe‹cient of the peak of cefaclor are not less than40,000 steps and 0.8 to 1.3, respectively.

System repeatability: When the test is repeated 3 timeswith 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviations of the peakareas and the retention times of cefaclor are not more than2.0z, respectively.

Water Not more than 6.5z (0.2 g, volumetric titration,back titration).

Assay Weigh accurately an amount of Cefaclor andCefaclor Reference Standard, equivalent to about 0.1 g(potency), and dissolve each in 0.1 mol/L phosphate buŠersolution, pH 4.5 to make exactly 100 mL. Pipet 10 mL eachof these solutions, add exactly 10 mL of the internal stan-dard solution, add 0.1 mol/L phosphate buŠer solution, pH4.5 to make 50 mL, and use these solutions as the samplesolution and the standard solution. Perform the test with10 mL each of the sample solution and the standard solutionas directed under the Liquid Chromatography according tothe following conditions, and determine the ratios, QT andQS, of the peak area of cefaclor to that of the internal stan-dard.

Amount [mg (potency)] of C15H14ClN3O4S

＝WS×QT

QS× 1000

WS: Amount [mg (potency)] of Cefaclor ReferenceStandard

Internal standard solution—A solution of 4-aminoacetophe-none in 0.1 mol/L phosphate buŠer solution, pH 4.5 (1 in700).Operating conditions—


Column: A stainless steel column 4.6 mm in inside di-ameter and 15 cm in length, packed with octadecylsilanizedsilica gel for liquid chromatography (5 mm in particle di-


ameter).Column temperature: A constant temperature of about

259C.Mobile phase: Dissolve 6.8 g of potassium dihydrogen

phosphate in 1000 mL of water, and adjust the pH to 3.4with diluted phosphoric acid (3 in 500). To 940 mL of thissolution add 60 mL of acetonitrile.

Flow rate: Adjust the ‰ow rate so that the retention timeof cefaclor is about 7 minutes.System suitability—

System performance: When the procedure is run with10 mL of the standard solution under the above operatingconditions, cefaclor and the internal standard are eluted inthis order with the resolution between these peaks being notless than 5.

System repeatability: When the test is repeated 6 timeswith 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratiosof the peak area of cefaclor to that of the internal standard isnot more than 1.0z.


Cefaloridinセファロリジン


Cefaloridin contains not less than 940 mg (potency)per mg, calculated on the anhydrous basis. The poten-cy of Cefaloridin is expressed as mass (potency) ofcefaloridin (C19H17N3O4S2).

Description Cefaloridin occurs as white to light yellowishwhite, crystals or crystalline powder.

It is soluble in water, slightly soluble in methanol, andvery slightly soluble in ethanol (95).

Identiˆcation (1) Determine the absorption spectrum ofa solution of Cefaloridin (1 in 50,000) as directed under theUltraviolet-visible Spectrophotometry, and compare thespectrum with the Reference Spectrum or the spectrum of asolution of Cefaloridin Reference Standard prepared in thesame manner as the sample solution: both spectra exhibitsimilar intensities of absorption at the same wavelengths.

(2) Determine the infrared absorption spectrum ofCefaloridin as directed in the potassium bromide diskmethod under the Infrared Spectrophotometry, and com-pare the spectrum with the Reference Spectrum or the spec-trum of Cefaloridin Reference Standard: both spectraexhibit similar intensities of absorption at the same wavenumbers.

(3) Determine the spectrum of a solution of Cefaloridinin heavy water for nuclear magnetic resonance spectroscopy(1 in 10) as directed under the Nuclear Magnetic Resonance

Spectroscopy (1H), using sodium 3-trimethylsilylpropanesul-fonate for nuclear magnetic resonance spectroscopy as aninternal reference compound: it exhibits a single signal A ataround d3.8 ppm, and two multiple signals, B and C,between d6.9 ppm and d7.5 ppm and between d7.9 ppm andd9.0 ppm, respectively. The ratio of the integrated intensityof each signal, A:B:C, is about 2:3:5.

Optical rotation [a]20D : ＋46 –＋519(0.5 g calculated on the


pH Dissolve 1.0 g of Cefaloridin in 10 mL of water: thepH of the solution is between 4.0 and 6.0.

Purity (1) Clarity and color of solution－Dissolve 1.0 gof Cefaloridin in 5 mL of water by warming: the solution isclear and light yellow.

(2) Heavy metals—Proceed with 1.0 g of Cefaloridinaccording to Method 2, and perform the test. Prepare thecontrol solution with 2.0 mL of Standard Lead Solution (notmore than 20 ppm).

(3) Arsenic—Prepare the test solution with 1.0 g ofCefaloridin according to Method 3, and perform the test(not more than 2 ppm).

(4) Related substances—Dissolve 20 mg of Cefaloridinin 20 mL of a solution of sodium dihydrogen phosphate di-hydrate (39 in 1250). To 2 mL of this solution add a solutionof sodium dihydrogen phosphate dihydrate (39 in 1250) tomake 20 mL, and use this solution as the sample solution.Perform the test with 20 mL of the sample solution as direct-ed under the Liquid Chromatography according to the fol-lowing conditions, determine each peak area by the auto-matic integration method, and calculate the amount of eachpeak by the area percentage method: the amount of eachpeak other than cefaloridin is not more than 0.5z, and thetotal amount of the peaks other than cefaloridin is not morethan 2.0z.Operating conditions—




Mobile phase: To 25.9 g of sodium acetate trihydrate add0.6 mL of acetic acid (100) and water to make 1000 mL. To900 mL of this solution add 50 mL of acetonitrile and 50 mLof methanol.

Flow rate: Adjust the ‰ow rate so that the retention timeof cefaloridin is about 14 minutes.

Time span of measurement: About 2 times as long as theretention time of cefaloridin after the solvent peak.System suitability—

Test for required detectability: Measure exactly 1 mL ofthe sample solution, and add a solution of sodium dihydro-gen phosphate dihydrate (39 in 1250) to make exactly100 mL. To exactly 1 mL of this solution add a solution ofsodium dihydrogen phosphate dihydrate (39 in 1250) to


make exactly 10 mL. Conˆrm that the peak area of cefalo-ridin obtained from 20 mL of this solution is equivalent to0.07 to 0.13z of that from 20 mL of the sample solution.

System performance: When the procedure is run with20 mL of the sample solution under the above operating con-ditions, the number of theoretical plates and the symmetrycoe‹cient of the peak of cefaloridin are not less than 5000steps and not more than 1.3, respectively.

System repeatability: When the test is repeated 6 timeswith 20 mL of the sample solution under the above operatingconditions, the relative standard deviation of the peak areaof cefaloridin is not more than 3.0z.


Assay Weigh accurately an amount of Cefaloridin andCefaloridin Reference Standard, equivalent to about 45 mg(potency), and dissolve each in water to make exactly 50 mL.Measure exactly 20 mL each of these solutions, add exactly5 mL of the internal standard solution and water to make100 mL, and use these solutions as the sample solution andthe standard solution. Perform the test with 5 mL each of thesample solution and the standard solution as directed underthe Liquid Chromatography according to the followingconditions, and determine the ratios, QT and QS, of the peakarea of cefaloridin to that of the internal standard.

Amount [mg (potency)] of C19H17N3O4S2

＝WS×QT

QS× 1000

WS: Amount [mg (potency)] of Cefaloridin ReferenceStandard

Internal standard solution—A solution of acetanilide inacetonitrile (9 in 5000).Operating conditions—




Mobile phase: Dissolve 25.9 g of sodium acetate trihy-drate in 0.6 mL of acetic acid (100) and water to make1000 mL. To 800 mL of this solution add 200 mL of acetoni-trile.

Flow rate: Adjust the ‰ow rate so that the retention timeof cefaloridin is about 5 minutes.System suitability—

System performance: When the procedure is run with 5 mLof the standard solution under the above operating condi-tions, cefaloridin and the internal standard are eluted in thisorder with the resolution between these peaks being not lessthan 1.5.

System repeatability: When the test is repeated 6 timeswith 5 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratios

of the peak area of cefaloridin to that of the internal stan-dard is not more than 1.0z.


Cefalotin Sodiumセファロチンナトリウム


Cefalotin Sodium contains not less than 910 mg(potency) per mg, calculated on the anhydrous basis.The potency of Cefalotin Sodium is expressed as mass(potency) of cefalotin (C16H16N2O6S2: 396.44).

Description Cefalotin Sodium occurs as white to light yel-lowish white, crystals or crystalline powder.

It is freely soluble in water, slightly soluble in methanol,very slightly soluble in ethanol (95), and practically insolublein acetonitrile.

Identiˆcation (1) Determine the absorption spectrum ofa solution of Cefalotin Sodium (1 in 50,000) as directed un-der the Ultraviolet-visible Spectrophotometry, and comparethe spectrum with the Reference Spectrum or the spectrumof a solution of Cefalotin Sodium Reference Standard pre-pared in the same manner as the sample solution: both spec-tra exhibit similar intensities of absorption at the same wave-lengths.

(2) Determine the infrared absorption spectrum ofCefalotin Sodium as directed in the potassium bromide diskmethod under the Infrared Spectrophotometry, and com-pare the spectrum with the Reference Spectrum or the spec-trum of Cefalotin Sodium Reference Standard: both spectraexhibit similar intensities of absorption at the same wavenumbers.

(3) Determine the spectrum of a solution of CefalotinSodium in heavy water for nuclear magnetic resonance spec-troscopy (1 in 10) as directed under the Nuclear MagneticResonance Spectroscopy (1H), using sodium 3-trimethylsilyl-propanesulfonate for nuclear magnetic resonance spec-troscopy as an internal reference compound: it exhibits a sin-gle signal A at around d2.1 ppm, a single or sharp multiplesignal B at around d3.9 ppm, and a multiple signal C ataround d7.0 ppm. The ratio of the integrated intensity ofthese signals, A:B:C, is about 3:2:2.

(4) Cefalotin Sodium responds to the Qualitative Test(1) for sodium salt.

Optical rotation [a]25D : ＋124 –＋1349(5 g, water, 100 mL,

100 mm).

pH The pH of a solution obtained by dissolving 1.0 g ofCefalotin Sodium in 10 mL of water is between 4.5 and 7.0.

Purity (1) Clarity and color of solution—Dissolve 1.0 gof Cefalotin Sodium in 5 mL of water: the solution is clear


and light yellow.(2) Heavy metals—Proceed with 1.0 g of Cefalotin

Sodium according to Method 2, and perform the test. Pre-pare the control solution with 2.0 mL of Standard LeadSolution (not more than 20 ppm).

(3) Arsenic—Prepare the test solution with 1.0 g ofCefalotin Sodium according to Method 3, and perform thetest (not more than 2 ppm).

(4) Related substances—Pipet 1 mL of the standardsolution obtained in the Assay, add the mobile phase tomake exactly 100 mL, and use this solution as the standardsolution. Perform the test with exactly 10 mL each of thesample solution obtained in the Assay and the standard solu-tion prepared here as directed under the Liquid Chro-matography according to the following conditions, and de-termine each peak area by the automatic integrationmethod: the peak area other than cefalotin from the samplesolution is not more than the peak area of cefalotin from thestandard solution, and the total area of the peaks other thancefalotin from the sample solution is not more than 3 timesthe peak area of cefalotin from the standard solution.Operating conditions—


Time span of measurement: About 4 times as long as theretention time of cefalotin.System suitability—

Test for required detectability: Measure exactly 1 mL ofthe standard solution, and add the mobile phase to makeexactly 10 mL. Conˆrm that the peak area of cefalotinobtained from 10 mL of this solution is equivalent to 7 to13z of that from 10 mL of the standard solution.

System performance: Heat the standard solution in awater bath of 909C for 10 minutes, and cool. Measureexactly 2.5 mL of this solution, and add the mobile phase tomake exactly 100 mL. When the procedure is run with 10 mLof this solution under the above operating conditions, theresolution between the peak of cefalotin and the peak, hav-ing the relative retention time of about 0.5 with respect tocefalotin, is not less than 9, and the symmetry coe‹cient ofthe peak of cefalotin is not more than 1.8.

System repeatability: When the test is repeated 3 timeswith 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peakarea of cefalotin is not more than 2.0z.


Assay Weigh accurately an amount of Cefalotin Sodiumand Cefalotin Sodium Reference Standard, equivalent toabout 25 mg (potency), and dissolve each in the mobilephase to make exactly 25 mL, and use these solutions as thesample solution and the standard solution. Perform the testwith exactly 10 mL each of the sample solution and the stan-dard solution as directed under the Liquid Chromatographyaccording to the following conditions, and determine thepeak areas, AT and AS, of cefalotin.

Amount [mg (potency)] of cefalotin (C16H16N2O6S2)

＝WS×AT

AS× 1000

WS: Amount [mg (potency)] of Cefalotin Sodium Refer-ence Standard





Mobile phase: Dissolve 17 g of sodium acetate trihydratein 790 mL of water, and add 0.6 mL of acetic acid (100). Ifnecessary adjust the pH to 5.9± 0.1 with 0.1 mol/L sodiumhydrochloride TS or acetic acid (100). To this solution add150 mL of acetonitrile and 70 mL of ethanol (95).

Flow rate: Adjust the ‰ow rate so that the retention timeof cefalotin is about 12 minutes.System suitability—

System performance: Heat the standard solution in awater bath of 909C for 10 minutes, and cool. Measureexactly 2.5 mL of this solution, and add the mobile phase tomake exactly 100 mL. When the procedure is run with 10 mLof this solution under the above operating conditions, theresolution between the peak of cefalotin and the peak, hav-ing the relative retention time of about 0.5 with respect tocefalotin is not less than 9, and the symmetry coe‹cient ofthe peak of cefalotin is not more than 1.8.

System repeatability: When the test is repeated 6 timeswith 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peakarea of cefalotin is not more than 1.0z.


Cefamandole Sodiumセファマンドールナトリウム


Cefamandole Sodium contains not less than 858 mg(potency) per mg, calculated on the anhydrous basis.The potency of Cefamandole Sodium is expressed asmass (potency) of cefamandole (C18H18N6O5S2:462.50).

Description Cefamandole Sodium occurs as a white tolight yellowish white crystalline powder.

It is very soluble in water, freely soluble in N,N-dimethyl-formamide, sparingly soluble in methanol, slightly soluble inethanol (99.5), and practically insoluble in acetonitrile.

It is hygroscopic.


Identiˆcation (1) Determine the absorption spectrum ofa solution of Cefamandole Sodium (1 in 50,000) as directedunder the Ultraviolet-visible Spectrophotometry, and com-pare the spectrum with the Reference Spectrum: both spec-tra exhibit similar intensities of absorption at the same wave-lengths.

(2) Determine the infrared absorption spectrum ofCefamandole Sodium as directed in the potassium bromidedisk method under the Infrared Spectrophotometry, andcompare the spectrum with the Reference Spectrum: bothspectra exhibit similar intensities of absorption at the samewave numbers.

(3) Determine the spectrum of a solution of Cefaman-dole Sodium in heavy water for nuclear magnetic resonancespectroscopy (1 in 10) as directed under the Nuclear Magnet-ic Resonance Spectroscopy (1H), using sodium 3-trimethyl-silylpropanesulfonate for nuclear magnetic resonance spec-troscopy as an internal reference compound: it exhibits twosingle signals, A and B, at around d4.0 ppm and at aroundd5.3 ppm, and a multiple signal C at around d7.5 ppm. Theratio of the integrated intensity of the two signals, B:C, isabout 1:5.

(4) Cefamandole Sodium responds to the QualitativeTest (1) for sodium salt.

Optical rotation [a]20D : －26.0 –－34.09(1 g calculated on


pH The pH of a solution obtained by dissolving 1.0 g ofCefamandole Sodium in 10 mL of water is between 4.3 and6.3.

Purity (1) Clarity and color of solution—Dissolve 0.5 gof Cefamandole Sodium in 2 mL of water: the solution isclear and pale yellow to light yellow.

(2) Heavy metals—Proceed with 1.0 g of CefamandoleSodium according to Method 4, and perform the test. Pre-pare the control solution with 2.0 mL of Standard LeadSolution (not more than 20 ppm).

(3) Arsenic—Prepare the test solution with 1.0 g ofCefamandole Sodium according to Method 5, and performthe test (not more than 2 ppm).

(4) 1-Methyl-1H-tetrazole-5-thiol—Dissolve 30 mg ofCefamandole Sodium in exactly 2 mL of the internal stan-dard solution, add water to make 20 mL, and use this solu-tion as the sample solution. Separately, dissolve 9 mg ofsodium 1-methyl-1H-tetrazole-5-thiolate in water to makeexactly 100 mL. Pipet 2 mL of this solution, add exactly2 mL of the internal standard solution and water to make20 mL, and use this solution as the standard solution. Per-form the test with 5 mL each of the sample solution and thestandard solution as directed under the Liquid Chro-matography according to the following conditions, and de-termine the ratios, QT and QS, of the peak area of 1-methyl-1H-tetrazole-5-thiol to that of the internal standard: QT isnot larger than QS.Internal standard solution—A solution of m-cresol (7 in10,000).





Mobile phase: Dissolve 6.94 g of potassium dihydrogenphosphate, 3.22 g of disodium hydrogen phosphate 12-waterand 1.60 g of tetra-n-butylammonium bromide in water tomake 1000 mL. To 600 mL of this solution add 200 mL ofmethanol.

Flow rate: Adjust the ‰ow rate so that the retention timeof 1-methyl-1H-tetrazole-5-thiol is about 4 minutes.System suitability—

System performance: When the procedure is run with 5 mLof the standard solution under the above operating condi-tions, 1-methyl-1H-tetrazole-5-thiol and the internal stan-dard are eluted in this order with the resolution betweenthese peaks being not less than 14.

System repeatability: When the test is repeated 3 timeswith 5 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratiosof the peak area of 1-methyl-1H-tetrazole-5-thiol to that ofthe internal standard is not more than 3.0z.

Water Not more than 2.0z (1 g, volumetric titration, backtitration).

Assay Weigh accurately an amount of CefamandoleSodium and Cefamandole Lithium Reference Standard,equivalent to about 50 mg (potency), and dissolve each in themobile phase to make exactly 100 mL. Measure exactly 2 mLeach of these solutions, add exactly 4 mL of the internalstandard solution and the mobile phase to make 20 mL, anduse these solutions as the sample solution and the standardsolution. Perform the test with 20 mL each of the samplesolution and the standard solution as directed under the Liq-uid Chromatography according to the following conditions,and determine the ratios, QT and QS, of the peak area ofcefamandole to that of the internal standard.

Amount [mg (potency)] of cefamandole (C18H18N6O5S2)

＝WS×QT

QS× 1000

WS: Amount [mg (potency)] of Cefamandole LithiumReference Standard

Internal standard solution—A solution of methyl para-hydroxybenzoate in the mobile phase (1 in 12,500).Operating conditions—


Column: A stainless steel column 4 mm in inside diameterand 15 cm in length, packed with divinylbenzene-methacry-late co-polymer for liquid chromatography (5 mm in particlediameter).

Column temperature: A constant temperature of about


259C.Mobile phase: Dissolve 15.0 g of disodium hydrogen

phosphate 12-water and 1.01 g of potassium dihydrogenphosphate in water to make 1000 mL, and add 1.6 g of tetra-n-butylammonium bromide. To 810 mL of this solution add290 mL of acetonitrile.

Flow rate: Adjust the ‰ow rate so that the retention timeof cefamandole is about 7 minutes.System suitability—

System performance: When the procedure is run with20 mL of the standard solution under the above operatingconditions, cefamandole and the internal standard are elutedin this order with the resolution between these peaks beingnot less than 3.

System repeatability: When the test is repeated 3 timeswith 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratiosof the peak area of cefamandole to that of the internal stan-dard is not more than 1.0z.


Cefbuperazone Sodiumセフブペラゾンナトリウム


Cefbuperazone Sodium contains not less than870 mg (potency) per mg, calculated on the anhydrousbasis. The potency of Cefbuperazone Sodium isexpressed as mass (potency) of cefbuperazone(C22H29N9O9S2: 627.65).

Description Cefbuperazone Sodium occurs as white tolight yellowish white, powder or masses.

It is very soluble in water, freely soluble in methanol andin pyridine, sparingly soluble in ethanol (95), and very slight-ly soluble in acetonitrile.

Identiˆcation (1) Determine the absorption spectrum ofa solution of Cefbuperazone Sodium (1 in 50,000) as direct-ed under the Ultraviolet-visible Spectrophotometry, andcompare the spectrum with the Reference Spectrum: bothspectra exhibit similar intensities of absorption at the samewavelengths.

(2) Dissolve 0.1 g of Cefbuperazone Sodium in 0.5 mLof deuterated pyridine for nuclear magnetic resonance spec-troscopy and 1 drop of heavy water for nuclear magneticresonance spectroscopy, and determine the spectrum of thissolution as directed under the Nuclear Magnetic ResonanceSpectroscopy (1H), using tetramethylsilane for nuclear mag-netic resonance spectroscopy as an internal reference com-pound: it exhibits a triplet signal A at around d1.1 ppm, andtwo doublet signals, B and C, at around d1.6 ppm and ataround d5.1 ppm, respectively. The ratio of the integrated

intensity of each signal, A:B:C, is about 3:3:1.(3) Cefbuperazone Sodium responds to the Qualitative

Test (1) for sodium salt.



pH Dissolve 1.0 g of Cefbuperazone Sodium in 4 mL ofwater: the pH of the solution is between 4.0 and 6.0.

Purity (1) Clarity and color of solution—Dissolve 1.0 gof Cefbuperazone Sodium in 4 mL of water: the solution isclear and light yellow.

(2) Heavy metals—Proceed with 2.0 g of CefbuperazoneSodium according to Method 4, and perform the test. Pre-pare the control solution with 2.0 mL of Standard LeadSolution (not more than 10 ppm).

(3) Arsenic—Prepare the test solution with 1.0 g of Cef-buperazone Sodium according to Method 4, and performthe test (not more than 2 ppm).

(4) Related substances—Dissolve 0.10 g of Cefbupera-zone Sodium in 100 mL of the mobile phase, and use thissolution as the sample solution. Pipet 1 mL of the samplesolution, add the mobile phase to make exactly 50 mL, anduse this solution as the standard solution. Perform the testwith exactly 25 mL each of the sample solution and the stan-dard solution as directed under the Liquid Chromatographyaccording to the following conditions, and determine eachpeak area by the automatic integration method. Calculatethe percentages of each peak area of related substances fromthe sample solution against 50 times of the peak area ofcefbuperazone from the standard solution; the amount ofrelated substance I having the relative retention time ofabout 0.2 to cefbuperazone is not more than 2.0z, theamount of related substance II having the relative retentiontime of about 0.6 to cefbuperazone is not more than 4.5z

and the amount of related substance III having the relativeretention time of about 1.6 to cefbuperazone is not morethan 1.0z, and the total amount of these related substancesis not more than 6.0z. For these calculations, use the valuesof the peak areas of the related substances I and III obtainedby the automatic integration method after multiplying byeach sensitivity coe‹cient, 0.72 and 0.69, respectively.Operating conditions—


Time span of measurement: About 2 times as long as theretention time of cefbuperazone.System suitability—

Test for required detectability: Measure exactly 1 mL ofthe standard solution, and add the mobile phase to makeexactly 10 mL. Conˆrm that the peak area of cefbuperazoneobtained from 25 mL of this solution is equivalent to 7 to13z of that from 25 mL of the standard solution.

System performance: When the procedure is run with25 mL of the standard solution under the above operatingconditions, the number of theoretical plates and the symmet-ry coe‹cient of the peak of cefbuperazone are not less than


5000 steps and not more than 1.5, respectively.System repeatability: When the test is repeated 6 times

with 25 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peakarea of cefbuperazone is not more than 2.0z.

Water Not more than 1.0z (3 g, volumetric titration,direct titration).

Assay Weigh accurately an amount of CefbuperazoneSodium and Cefbuperazone Reference Standard, equivalentto about 0.1 g (potency), and dissolve each in the mobilephase to make exactly 100 mL. Measure exactly 10 mL eachof these solutions, add exactly 10 mL of the internal stan-dard solution and the mobile phase to make 50 mL, and usethese solutions as the sample solution and the standard solu-tion. Perform the test with 10 mL each of the sample solutionand the standard solution as directed under the LiquidChromatography according to the following conditions, anddetermine the ratios, QT and QS, of the peak area of cef-buperazone to that of the internal standard.

Amount [mg (potency)] of cefbuperazone (C22H29N9O9S2)

＝WS×QT

QS× 1000

WS: Amount [mg (potency)] of Cefbuperazone ReferenceStandard

Internal standard solution—A solution of acetonitrile in themobile phase (1 in 4000).Operating conditions—




Mobile phase: Dissolve 2.0 g of tetra-n-propylammoniumbromide in 1000 mL of a mixture of water, acetonitrile andacetic acid-sodium acetate buŠer solution, pH 5.0 (83:13:4).

Flow rate: Adjust the ‰ow rate so that the retention timeof cefbuperazone is about 16 minutes.System suitability—

System performance: When the procedure is run with10 mL of the standard solution under the above operatingconditions, the internal standard and cefbuperazone areeluted in this order with the resolution between these peaksbeing not less than 3.

System repeatability: When the test is repeated 6 timeswith 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratiosof the peak area of cefbuperazone to that of the internalstandard is not more than 1.0z.

Containers and storage Containers—Hermetic containers.Storage—In a cold place.

Cefmenoxime Hydrochloride塩酸セフメノキシム


Cefmenoxime Hydrochloride contains not less than890 mg (potency) per mg, calculated on the dehydratedbasis. The potency of Cefmenoxime Hydrochlorideis expressed as mass (potency) of cefmenoxime(C16H17N9O5S3: 511.57).

Description Cefmenoxime Hydrochloride occurs as whiteto light orange-yellow crystals or crystalline powder.

It is freely soluble in formamide and in dimethylsulfoxide,slightly soluble in methanol, very slightly soluble in water,and practically insoluble in ethanol (95).

Identiˆcation (1) Determine the absorption spectrum ofa solution of Cefmenoxime Hydrochloride in 0.1 mol/Lphosphate buŠer solution, pH 6.8 (3 in 200,000) as directedunder the Ultraviolet-visible Spectrophotometry, and com-pare the spectrum with the Reference Spectrum or the spec-trum of a solution of Cefmenoxime Hydrochloride Refer-ence Standard prepared in the same manner as the samplesolution: both spectra exhibit similar intensities of absorp-tion at the same wavelengths.

(2) Determine the infrared absorption spectrum ofCefmenoxime Hydrochloride as directed in the potassiumbromide disk method under the Infrared Spectrophotomet-ry, and compare the spectrum with the Reference Spectrumor the spectrum of Cefmenoxime Hydrochloride ReferenceStandard: both spectra exhibit similar intensities of absorp-tion at the same wave numbers.

(3) Determine the spectrum of a solution of Cef-menoxime Hydrochloride in deuterated dimethylsulfoxidefor nuclear magnetic resonance spectroscopy (1 in 10) asdirected under the Nuclear Magnetic Resonance Spec-troscopy (1H), using tetramethylsilane for nuclear magneticresonance spectroscopy as an internal reference compound:it exhibits two single signals, A and B, at around d3.9 ppm,and a single signal C at around d6.8 ppm. The ratio of theintegrated intensity of each signal, A:B:C, is about 3:3:1.

(4) Dissolve 0.01 g of Cefmenoxime Hydrochloride in1 mL of diluted sodium carbonate TS (1 in 20), add 5 mL ofacetic acid (100) and 2 drops of silver nitrate TS: a whiteprecipitate is formed.

Optical rotation [a]20D : －27 –－359(1 g, 0.1 mol/L phos-

phate buŠer solution, pH 6.8, 100 mL, 100 mm).

pH The pH of a solution obtained by dissolving 0.10 g ofCefmenoxime Hydrochloride in 150 mL of water is between2.8 and 3.3.

Purity (1) Clarity and color of solution—A solution ob-tained by dissolving 1.0 g of Cefmenoxime Hydrochloride in10 mL of diluted sodium carbonate TS (1 in 4) is clear andcolorless to light yellow.


(2) Heavy metals—Proceed with 1.0 g of CefmenoximeHydrochloride according to Method 4, and perform the test.Prepare the control solution with 2.0 mL of Standard LeadSolution (not more than 20 ppm).

(3) Arsenic—Prepare the test solution by incinerating1.0 g of Cefmenoxime Hydrochloride according to Method4 and adding 10 mL of dilute hydrochloric acid to theresidue after cooling, and perform the test (not more than2 ppm).

(4) Related substances—Weigh accurately about 0.1 gof Cefmenoxime Hydrochloride, dissolve in 20 mL of0.1 mol/L phosphate buŠer solution, pH 6.8, and add themobile phase to make exactly 50 mL. Pipet 4 mL of thissolution, add the mobile phase to make exactly 50 mL, anduse this solution as the sample solution. Separately, weighaccurately about 10 mg of 1-methyl-1H-tetrazol-5-thiol, anddissolve in the mobile phase to make exactly 100 mL. Pipet4 mL of this solution, add the mobile phase to make exactly250 mL, and use this solution as the standard solution (1).Weigh accurately about 0.1 g of Cefmenoxime Hydrochlo-ride Reference Standard, dissolve in 20 mL of 0.1 mol/Lphosphate buŠer solution, pH 6.8, and add the mobile phaseto make exactly 100 mL. Pipet 1 mL of this solution, add themobile phase to make exactly 250 mL, and use this solutionas the standard solution (2). Perform the test immediatelyafter preparation of these solutions with exactly 10 mL eachof the sample solution, the standard solution (1) and thestandard solution (2) as directed under the Liquid Chro-matography according to the following conditions. Deter-mine each peak area obtained from the chromatograms ofthese solutions by the automatic integration method, andcalculate the amounts of 1-methyl-1H-tetrazol-5-thiol andthe total related substance by the following formula: theamount of 1-methyl-1H-tetrazol-5-thiol is not more than1.0z, and the total related substance is not more than 3.0z.

Amount (z) of 1-methyl-1H-tetrazol-5-thiol

＝WSa

WT×

ATa

ASa× 20

Amount (z) of total related substance

＝WSa

WT×

ATa

ASa× 20＋

WSb

WT＋

ST

ASb× 5

WSa: Amount (g) of 1-methyl-1H-tetrazol-5-thiolWSb: Amount (g) of Cefmenoxime Hydrochloride Refer-

ence StandardWT: Amount (g) of the sampleASa: Peak area of 1-methyl-1H-tetrazol-5-thiol from the

standard solution (1)ASb: Peak area of cefmenoxime from the standard solu-

tion (2)ATa: Peak area of 1-methyl-1H-tetrazol-5-thiol from the

sample solutionST: Total area of the peaks other than 1-methyl-1H-

tetrazol-5-thiol and other than cefmenoxime from thesample solution

Operating conditions—Detector, column, column temperature, mobile phase,

and ‰ow rate: Proceed as directed in the operating condi-tions in the Assay.

Time span of measurement: About 2.5 times as long as theretention time of cefmenoxime.System suitability—

Test for required detectability: Measure exactly 5 mL ofthe standard solution (1), add the mobile phase to makeexactly 100 mL. Conˆrm that the peak area of 1-methyl-1H-tetrazol-5-thiol obtained from 10 mL of this solution isequivalent to 4.5 to 5.5z of that from the standard solution(1). Then, measure exactly 2 mL of the standard solution (2),add the mobile phase to make exactly 100 mL. Conˆrm thatthe peak area of cefmenoxime obtained from 10 mL of thissolution is equivalent to 1.5 to 2.5z of that from the stan-dard solution (2).


System repeatability: When the test is repeated 6 timeswith 10 mL of the standard solution (1) under the aboveoperating conditions, the relative standard deviation of thepeak area of 1-methyl-1H-tetrazol-5-thiol is not more than1.0z.

Water Not more than 1.5z (1 g, volumetric titration,direct titration. Use a mixture of formamide for Karl Fishermethod and methanol for Karl Fisher method (2:1)).

Assay Weigh accurately an amount of CefmenoximeHydrochloride and Cefmenoxime Hydrochloride ReferenceStandard, equivalent to about 50 mg (potency), dissolveeach in 10 mL of 0.1 mol/L phosphate buŠer solution, pH6.8, and add the mobile phase to make exactly 50 mL. Pipet4 mL each of these solutions, add exactly 20 mL of the inter-nal standard solution and the mobile phase to make 50 mL,and use these solutions as the sample solution and the stan-dard solution, respectively. Perform the test with 10 mL eachof the sample solution and the standard solution as directedunder the Liquid Chromatography according to the follow-ing conditions, and determine the ratios, QT and QS, of thepeak area of cefmenoxime to that of the internal standard.

Amount [mg (potency)] of cefmenoxime (C16H17N9O5S3)

＝WS×QT

QS× 1000

WS: Amount [mg (potency)] of Cefmenoxime Hydrochlo-ride Reference Standard

Internal standard solution—A solution of phthalimide inmethanol (3 in 2000).Operating conditions—




Mobile phase: A mixture of water, acetonitrile and aceticacid (100) (50:10:1).


Flow rate: Adjust the ‰ow rate so that the retention timeof cefmenoxime is about 8 minutes.System suitability—

System performance: When the procedure is run with10 mL of the standard solution under the above operatingconditions, cefmenoxime and the internal standard are elut-ed in this order with the resolution between these peaksbeing not less than 2.3.

System repeatability: When the test is repeated 6 timeswith 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratiosof the peak areas of cefmenoxime to that of the internalstandard is not more than 1.0z.

Containers and storage Containers—Hermetic containers.

Add the following:

Cefodizime Sodiumセフォジジムナトリウム

C20H18N6Na2O7S4: 628.63Disodium (6R,7R )-7-[(Z )-2-(2-aminothiazol-4-yl)-2-(methoxyimino)acetylamino]-3-[(5-carboxylatomethyl-4-methylthiazol-2-yl)sulfanylmethyl]-8-oxo-5-thia-1-azabicycle[4.2.0]oct-2-ene-2-carboxylate [86329-79-5]

Cefodizime Sodium contains not less than 890 mg(potency) per mg, calculated on the anhydrous basisand corrected by the ethanol amount. The potency ofCefodizime Sodium is expressed as mass (potency) ofcefodizime (C20H20N6O7S4: 584.67).

Description Cefodizime Sodium occurs as a white to lightyellowish white crystalline powder.

It is very soluble in water, and practically insoluble inacetonitrile and in ethanol (99.5).

Identiˆcation (1) Determine the absorption spectrum ofa solution of Cefodizime Sodium (1 in 50,000) as directedunder the Ultraviolet-visible Spectrophotometry, and com-pare the spectrum with the Reference Spectrum or the spec-trum of a solution of Cefodizime Sodium Reference Stan-dard prepared in the same manner as the sample solution:both spectra exhibit similar intensities of absorption at thesame wavelengths.

(2) Determine the infrared absorption spectrum ofCefodizime Sodium as directed in the potassium bromidedisk method under the Infrared Spectrophotometry, andcompare the spectrum with the Reference Spectrum or thespectrum of Cefodizime Sodium Reference Standard: both

spectra exhibit similar intensities of absorption at the samewave numbers.

(3) Determine the spectrum of a solution of CefodizimeSodium in heavy water for nuclear magnetic resonance spec-troscopy (1 in 10) as directed under the Nuclear MagneticResonance Spectroscopy (1H), using sodium 3-trimethylsilyl-propanesulfonate for nuclear magnetic resonance spec-troscopy as an internal reference compound: it exhibitssingle signals, A, B and C, at around d2.3 ppm, at aroundd4.0 ppm, and at around d7.0 ppm. The ratio of the in-tegrated intensity of these signals, A:B:C, is about 3:3:1.

(4) Cefodizime Sodium responds to the Qualitative Test(1) for sodium salt.

Optical rotation [a]20D : －56 –－629(0.2 g calculated on the

anhydrous basis and corrected by the ethanol amount,water, 20 mL, 100 mm).

pH Dissolve 1.0 g of Cefodizime Sodium in 10 mL ofwater: the pH of the solution is between 5.5 and 7.5.

Purity (1) Clarity and color of solution—Dissolve 1.0 gof Cefodizime Sodium in 10 mL of water: the solution isclear and pale yellow to light yellow.

(2) Heavy metals—Weigh 1.0 g of Cefodizime Sodiumin a crucible, cover loosely, and carbonize by gentle heating.After cooling, add 2 mL of sulfuric acid, heat gradually un-til the white fumes are no longer evolved, and ignite between5009C and 6009C. Proceed according to Method 2, and per-form the test. Prepare the control solution with 2.0 mL ofStandard Lead Solution (not more than 20 ppm).

(3) Arsenic—Prepare the test solution with 1.0 g ofCefodizime Sodium according to Method 3, and perform thetest (not more than 2 ppm).

(4) Related substances—Dissolve 30 mg of CefodizimeSodium in 10 mL of the mobile phase, and use this solutionas the sample solution. Pipet 1 mL of the sample solution,add the mobile phase to make exactly 100 mL, and use thissolution as the standard solution. Perform the test with ex-actly 5 mL each of the sample solution and the standard solu-tion as directed under the Liquid Chromatography accord-ing to the following conditions, and determine each peakarea by the automatic integration method: the peak areaother than cefodizime from the sample solution is not morethan the peak area of cefodizime from the standard solution,and the total area of the peaks other than cefodizime fromthe sample solution is not more than 3 times the peak area ofcefodizime from the standard solution.Operating conditions—


Time span of measurement: About 4 times as long as theretention time of cefodizime after the solvent peak.System suitability—

Test for required detectability: Measure exactly 2 mL ofthe standard solution, and add the mobile phase to makeexactly 20 mL. Conˆrm that the peak area of cefodizimeobtained from 5 mL of this solution is equivalent to 7 to 13z


of that from 5 mL of the standard solution.System performance, and system repeatability: Proceed as

directed in the system suitability in the Assay.(5) Ethanol—Weigh accurately about 1 g of Cefodizime

Sodium, and dissolve in water to make exactly 10 mL. Pipet2 mL of this solution, add exactly 2 mL of the internal stan-dard solution, and use this solution as the sample solution.Separately, weigh accurately about 2 g of ethanol for gaschromatography, and add water to make exactly 1000 mL.Pipet 2 mL of this solution, add exactly 2 mL of the internalstandard solution, and use this solution as the standard solu-tion. Perform the test with 10 mL each of the sample solutionand the standard solution as directed under the Gas Chro-matography according to the following conditions, and cal-culate the ratios, QT and QS, of the peak area of ethanol tothat of the internal standard: the amount of ethanol is notmore than 2.0z.

Amount (z) of ethanol＝WS

WT×

QT

QS

WS: Amount (g) of ethanol for gas chromatographyWT: Amount (g) of the sample

Internal standard solution—A solution of 1-propanol (1 in400).Operating conditions—


3 m in length, packed with tetra‰uoroethylene polymer forgas chromatography (180 – 250 mm in particle diameter)coated in 15z with polyethylene glycol 20 M.



of ethanol is about 3 minutes.System suitability—

System performance: When the procedure is run with10 mL of the standard solution under the above operatingconditions, ethanol and the internal standard are eluted inthis order with the resolution between these peaks being notless than 2.5.

System repeatability: When the test is repeated 6 timeswith 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratiosof the peak area of ethanol to that of the internal standard isnot more than 2.0z.


Assay Weigh accurately an amount of Cefodizime Sodiumand Cefodizime Sodium Reference Standard, equivalent toabout 50 mg (potency), add exactly 10 mL of the internalstandard solution to dissolve, add water to make 100 mL,and use these solutions as the sample solution and the stan-dard solution. Perform the test with 10 mL each of the sam-ple solution and the standard solution as directed under theLiquid Chromatography according to the following condi-

tions, and determine the ratios, QT and QS, of the peak areaof cefodizime to that of the internal standard.

Amount [mg (potency)] of cefodizime (C20H20N6O7S4)

＝WS×QT

QS× 1000

WS: Amount [mg (potency)] of Cefodizime Sodium Ref-erence Standard

Internal standard solution—A solution of anhydrouscaŠeine (3 in 400).Operating conditions—




Mobile phase: Dissolve 0.80 g of potassium dihydrogenphosphate and 0.20 g of anhydrous disodium hydrogenphosphate in a suitable amount of water, and add 80 mL ofacetonitrile and water to make 1000 mL.

Flow rate: Adjust the ‰ow rate so that the retention timeof cefodizime is about 5 minutes.System suitability—

System performance: When the procedure is run with10 mL of the standard solution under the above operatingconditions, cefodizime and the internal standard are elutedin this order with the resolution between these peaks beingnot less than 6.

System repeatability: When the test is repeated 6 timeswith 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratiosof the peak area of cefodizime to that of the internal stan-dard is not more than 2.0z.


Cefotaxime Sodiumセフォタキシムナトリウム


Cefotaxime Sodium contains not less than 916 mg(potency) per mg, calculated on the dried basis. Thepotency of Cefotaxime Sodium is expressed as mass(potency) of cefotaxime (C16H17N5O7S2: 455.47).

Description Cefotaxime Sodium occurs as white to lightyellowish white crystalline powder.

It is freely soluble in water, sparingly soluble in methanol,and very slightly soluble in ethanol (95).

Identiˆcation (1) Dissolve 2 mg of Cefotaxime Sodium in0.01 mol/L hydrochloric acid TS to make 100 mL. Deter-


mine the absorption spectrum of this solution as directed un-der the Ultraviolet-visible Spectrophotometry, and comparethe spectrum with the Reference Spectrum: both spectraexhibit similar intensities of absorption at the same wave-lengths.

(2) Determine the infrared absorption spectrum ofCefotaxime Sodium as directed in the potassium bromidedisk method under the Infrared Spectrophotometry, andcompare the spectrum with the Reference Spectrum: bothspectra exhibit similar intensities of absorption at the samewave numbers.

(3) Determine the spectrum of a solution of CefotaximeSodium in heavy water for nuclear magnetic resonance spec-troscopy (1 in 125) as directed under the Nuclear MagneticResonance Spectroscopy (1H), using sodium 3-trimethylsilyl-propanesulfonate for nuclear magnetic resonance spec-troscopy as an internal reference compound: it exhibits threesingle signals, A, B and C, at around d2.1 ppm, at aroundd4.0 ppm and at around d7.0 ppm. The ratio of the integrat-ed intensity of each signal, A:B:C, is about 3:3:1.

(4) Cefotaxime Sodium responds to the Qualitative Test(1) for sodium salt.

Optical rotation [a]20D :＋58 –＋649(0.25 g calculated on the


pH The pH of a solution obtained by dissolving 1.0 g ofCefotaxime Sodium in 10 mL of water is between 4.5 and6.5.

Purity (1) Clarity and color of solution—A solutionobtained by dissolving 1.0 g of Cefotaxime Sodium in 10 mLof water is clear and light yellow.

(2) Sulfate—Dissolve 2.0 g of Cefotaxime Sodium in40 mL of water, add 2 mL of dilute hydrochloric acid andwater to make 50 mL, shake well, and ˆlter. Discard ˆrst10 mL of the ˆltrate, and to the subsequent 25 mL of theˆltrate add water to make 50 mL. Perform the test with thissolution as the test solution. Prepare the control solution asfollows: To 1.0 mL of 0.005 mol/L sulfuric acid VS add1 mL of dilute hydrochloric acid and water to make 50 mL(not more than 0.048z).

(3) Heavy metals—Proceed with 1.0 g of CefotaximeSodium according to Method 2, and perform the test. Pre-pare the control solution with 2.0 mL of Standard LeadSolution (not more than 20 ppm).

(4) Arsenic—Prepare the test solution with 1.0 g ofCefotaxime Sodium according to Method 3, and performthe test (not more than 2 ppm).

(5) Related substances—Perform the test with 10 mL ofthe sample solution obtained in the Assay as directed underthe Liquid Chromatography according to the following con-ditions, and determine each peak area obtained from thechromatogram by the automatic integration method: theeach peak area other than cefotaxime is not more than 1.0z

and the total of these peak areas is not more than 3.0z.Operating conditions—

Detector, column, column temperature, mobile phase A,mobile phase B, ‰owing of the mobile phase, and ‰ow rate:

Proceed as directed in the operating conditions in the Assay.Time span of measurement: About 3.5 times as long as the

retention time of cefotaxime after the solvent peak.System suitability—


Test for required detectability: Measure exactly 2 mL ofthe standard solution, and add the mobile phase A to makeexactly 100 mL. Pipet 2 mL of this solution, and add themobile phase A to make exactly 20 mL. Conˆrm that thepeak area of cefotaxime obtained from 10 mL of this solu-tion is equivalent to 0.15 to 0.25z of that obtained from10 mL of the standard solution.


Assay Weigh accurately an amount of Cefotaxime Sodiumand Cefotaxime Reference Standard, equivalent to about40 mg (potency), dissolve each in the mobile phase A tomake exactly 50 mL, and use these solutions as the samplesolution and the standard solution. Perform the test withexactly 10 mL each of the sample solution and the standardsolution as directed under the Liquid Chromatography ac-cording to the following conditions, and determine the peakareas, AT and AS, of cefotaxime of these solutions.

Amount [mg (potency)] of cefotaxime (C16H17N5O7S2)

＝WS×AT

AS× 1000

WS: Amount [mg (potency)] of Cefotaxime ReferenceStandard





Mobile phase A: To 0.05 mol/L disodium hydrogenphosphate TS add phosphoric acid to adjust the pH to 6.25.To 860 mL of this solution add 140 mL of methanol.

Mobile phase B: To 0.05 mol/L disodium hydrogenphosphate TS add phosphoric acid to adjust the pH to 6.25.To 600 mL of this solution add 400 mL of methanol.



Mobile phaseA (z)

Mobile phaseB (z)

0 – 7 100 07 – 9 100→ 80 0→ 209 – 16 80 20

16 – 45 80→ 0 20→ 10045 – 50 0 100


Flow rate: Adjust the ‰ow rate so that the retention timeof cefotaxime is about 14 minutes (about 1.3 mL/min).System suitability—

System performance: To 1 mL of the standard solutionadd 7.0 mL of water and 2.0 mL of methanol, mix, then add25 mg of sodium carbonate decahydrate, and shake. Afterallowing to stand for 10 minutes, add 3 drops of acetic acid(100) and 1 mL of the standard solution, and mix. When theprocedure is run with 10 mL of this solution under the aboveoperating conditions, desacetyl cefotaxime with the relativeretention time being about 0.3 to cefotaxime and cefotaximeare eluted in this order with the resolution between thesepeaks being not less than 20, and the symmetry coe‹cient ofthe peak of cefotaxime is not more than 2.

System repeatability: When the test is repeated 6 timeswith 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peakarea of cefotaxime is not more than 2.0z.


Cefotetanセフォテタン


Cefotetan contains not less than 960 mg (potency)per mg, calculated on the anhydrous basis. Thepotency of Cefotetan is expressed as mass (potency) ofcefotetan (C17H17N7O8S4).

Description Cefotetan occurs as white to light yellowishwhite powder.

It is sparingly soluble in methanol, and slightly soluble inwater and in ethanol (99.5).

Identiˆcation (1) Determine the absorption spectrum ofa solution of Cefotetan in phosphate buŠer solution forantibiotics, pH 6.5 (1 in 100,000) as directed under theUltraviolet-visible Spectrophotometry, and compare thespectrum with the Reference Spectrum or the spectrum of asolution of Cefotetan Reference Standard prepared in thesame manner as the sample solution: both spectra exhibitsimilar intensities of absorption at the same wavelengths.

(2) Determine the infrared absorption spectrum ofCefotetan as directed in the potassium bromide disk methodunder the Infrared Spectrophotometry, and compare thespectrum with the Reference Spectrum or the spectrum ofCefotetan Reference Standard: both spectra exhibit similarintensities of absorption at the same wavelengths.

(3) Dissolve 50 mg of Cefotetan in 0.5 mL of a solutionof sodium hydrogen carbonate in heavy water for nuclearmagnetic resonance spectroscopy (1 in 25). Determine thespectrum of this solution as directed under the NuclearMagnetic Resonance Spectroscopy (1H), using sodium3-trimethylsilylpropanesulfonate for nuclear magnetic

resonance spectroscopy as an internal reference compound:it exhibits single signals, A, B, C and D, at around d3.6 ppm, at around d4.0 ppm, at around d5.1 ppm and ataround d5.2 ppm, respectively. The ratio of the integratedintensity of each signal, A:B:C:D, is about 3:3:1:1.


the anhydrous basis, a solution of sodium hydrogen car-bonate (1 in 200), 50 mL, 100 mm).

Purity (1) Clarity and color of solution—Dissolve 1.0 gof Cefotetan in 10 mL of a solution of sodium hydrogencarbonate (1 in 30): the solution is clear, and colorless orlight yellow.

(2) Heavy metals—Proceed with 1.0 g of Cefotetanaccording to Method 2, and perform the test. Prepare thecontrol solution with 2.0 mL of Standard Lead Solution (notmore than 20 ppm).

(3) Related substances—Weigh accurately about 0.1 g ofCefotetan, dissolve in a suitable amount of methanol, addexactly 2 mL of the internal standard solution and methanolto make 20 mL, and use this solution as the sample solution.Separately, weigh accurately about 3 mg of 1-methyl-1H-tetrazole-5-thiol for liquid chromatography, previouslydried in a desiccator (in vacuum, silica gel) for 2 hours, andabout 2 mg of Cefotetan Reference Standard, calculated onthe anhydrous basis, dissolve in methanol to make exactly 20mL. Pipet 2 mL of this solution, add exactly 2 mL of the in-ternal standard solution and methanol to make 20 mL, anduse this solution as the standard solution. Perform the testwith 5 mL of the sample solution and the standard solutionas directed under the Liquid Chromatography according tothe following conditions, and determine the ratios, QTa, QTb,QTc, QTd, QTe and QTf, of the peak areas of 1-methyl-1H-tetrazole-5-thiol, cefotetan lactone having the relative reten-tion time of about 0.5 with respect to cefotetan, D2-cefote-tan having the relative retention time of about 1.2 withrespect to cefotetan, isothiazole substance having the rela-tive retention time of about 1.3 with respect to cefotetan,each of other related substances and the total of other relat-ed substances, to the peak area of the internal standard, re-spectively, obtained from the sample solution, and the ra-tios, QSa and QSb, of the peak areas of 1-methyl-1H-tetrazole-5-thiol and cefotetan, to the peak area of the inter-nal standard, respectively, obtained from the standard solu-tion. Calculate the amount of 1-methyl-1H-tetrazole-5-thiol,cefotetan lactone, D2-cefotetan, isothiazole substance, eachof other related substances and the total of other related sub-stances from the following equations: the amount of 1-methyl-1H-tetrazole-5-thiol is not more than 0.3z, cefote-tan lactone is not more than 0.3z, D2-cefotetan is not morethan 0.5z, isothiazole substance is not more than 0.5z,each of other related substances is not more than 0.2z andthe total of other related substances is not more than 0.4z.


1-Methyl-1H-tetrazole-5-thiol (z)＝WSa

WT×

QTa

QSa×

1100

Cefotetan lactone (z)＝WSb

WT×

QTb

QSb×

1100

D2-Cefotetan (z)＝WSb

WT×

QTc

QSb×

1100

Isothiazole substance (z)＝WSb

WT×

QTd

QSb×

1100

Each of other related substances (z)＝WSb

WT×

QTe

QSb×

1100

Total of other related substances (z)＝WSb

WT×

QTf

QSb×

1100

WSa: Amount (mg) of 1-methyl-1H-tetrazole-5-thiolWSb: Amount (mg) of Cefotetan Reference Standard, cal-

culated on the anhydrous basisWT: Amount (g) of the sample

Internal standard solution—A solution of anhydrouscaŠeine in methanol (3 in 10,000).Operating conditions—


Time span of measurement: About 3.5 times as long as theretention time of cefotetan.System suitability—

Test for required detectability: Measure exactly 15 mL ofthe standard solution, and add methanol to make exactly100 mL. Conˆrm that the peak area of cefotetan obtainedfrom 5 mL of this solution is equivalent to 12 to 18z of thatfrom 5 mL of the standard solution.


System repeatability: When the test is repeated 6 timeswith 5 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio ofthe peak area of cefotetan to that of the internal standard isnot more than 2.0z.



Isomer ratio Dissolve 10 mg of Cefotetan in 20 mL ofmethanol, and use this solution as the sample solution. Per-form the test with 5 mL of the sample solution as directed un-der the Liquid Chromatography according to the followingconditions, and determine each peak area by the automaticintegration method. Calculate the amount of the adjacenttwo peaks appeared at around the retention time of 40minutes, one having shorter retention time is l-substance andanother having longer retention time is d-substance, by thearea percentage method: the amount of l-substance is notless than 35z and not more than 45z.Operating conditions—


Column: A stainless steel column 4 mm in inside diameter



Mobile phase: A mixture of 0.1 mol/L phosphate buŠersolution, pH 7.0, water and a solution of tetrabutylammoni-um hydrogensulfate in acetonitrile (1 in 150) (9:9:2).

Flow rate: Adjust the ‰ow rate so that the retention timeof l-substance is about 40 minutes.System suitability—

System performance: When the procedure is run with 5 mLof the sample solution under the above operating conditions,l-substance and d-substance are eluted in this order with theresolution between these peaks being not less than 1.5.

System repeatability: To exactly 1 mL of the sample solu-tion add methanol to make exactly 10 mL. When the test isrepeated 6 times with 5 mL of this solution under the aboveoperating conditions, the relative standard deviation of thepeak area of l-substance is not more than 5z.

Assay Weigh accurately an amount of Cefotetan andCefotetan Reference Standard, equivalent to about 50 mg(potency), and dissolve each in phosphate buŠer solution forantibiotics, pH 6.5 to make exactly 50 mL. Pipet 15 mL eachof these solutions, add exactly 10 mL of the internal stan-dard solution and phosphate buŠer solution for antibiotics,pH 6.5 to make 50 mL, and use these solutions as the samplesolution and the standard solution. Perform the test with5 mL each of the sample solution and the standard solutionas directed under the Liquid Chromatography according tothe following conditions, and determine the ratios, QT andQS, of the peak area of cefotetan to that of the internalstandard.


＝WS×QT

QS× 1000

WS: Amount [mg (potency)] of Cefotetan Reference Stan-dard

Internal standard solution—A solution of anhydrouscaŠeine (1 in 1000).Operating conditions—




Mobile phase: Dissolve 11.53 g of phosphoric acid in1000 mL of water. To 850 mL of this solution add 50 mL ofacetonitrile, 50 mL of acetic acid (100) and 50 mL ofmethanol.

Flow rate: Adjust the ‰ow rate so that the retention timeof cefotetan is about 17 minutes.System suitability—

System performance: When the procedure is run with 5 mL


of the standard solution under the above operating condi-tions, the internal standard and cefotetan are eluted in thisorder with the resolution between these peaks being not lessthan 8.

System repeatability: When the test is repeated 5 timeswith 5 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio ofthe peak area of cefotetan to that of the internal standard isnot more than 1.0z.

Containers and storage Containers—Tight containers.Storage—Light-resistant, and at a temperature not ex-

ceeding 59C.

Cefotiam Hexetil Hydrochloride塩酸セフォチアムヘキセチル


Cefotiam Hexetil Hydrochloride contains not lessthan 615 mg (potency) per mg, calculated on the anhy-drous basis. The potency of Cefotiam HexetilHydrochloride is expressed as mass (potency) ofcefotiam (C18H23N9O4S3: 525.63).

Description Cefotiam Hexetil Hydrochloride occurs as awhite to light yellow powder.

It is very soluble in water, in methanol and in ethanol (95),freely soluble in dimethylsulfoxide, and slightly soluble inacetonitrile.

It dissolves in 0.1 mol/L hydrochloric acid TS.It is hygroscopic.

Identiˆcation (1) Determine the absorption spectrum ofa solution of Cefotiam Hexetil Hydrochloride in 0.1 mol/Lhydrochloric acid TS (3 in 125,000) as directed under theUltraviolet-visible Spectrophotometry, and compare thespectrum with the Reference Spectrum or the spectrum of asolution of Cefotiam Hexetil Hydrochloride ReferenceStandard prepared in the same manner as the sample solu-tion: both spectra exhibit similar intensities of absorption atthe same wavelengths.

(2) Determine the spectrum of a solution of CefotiamHexetil Hydrochloride in deuterated dimethylsulfoxide fornuclear magnetic resonance spectroscopy (1 in 20) as direct-ed under the Nuclear Magnetic Resonance Spectroscopy(1H), using tetramethylsilane for nuclear magnetic resonancespectroscopy as an internal reference compound: it exhibitstwo single signals, A and B, at around d2.8 ppm and ataround d6.6 ppm, and a multiple signal, C, at around d6.9ppm. The ratio of the integrated intensity of each signal,A:B:C, is about 6:1:1.

(3) To a solution of Cefotiam Hexetil Hydrochloride(1 in 200) add 2 mL of dilute nitric acid and 1 mL of silvernitrate TS, and mix: a white precipitate is formed.


anhydrous basis, 0.1 mol/L hydrochloric acid TS, 10 mL,100 mm).

Purity (1) Heavy metals—Proceed with 2.0 g of Cefo-tiam Hexetil Hydrochloride according to Method 2, andperform the test. Prepare the control solution with 2.0 mLof Standard Lead Solution (not more than 10 ppm).

(2) Arsenic—Prepare the test solution with 2.0 g ofCefotiam Hexetil Hydrochloride according to Method 3,and perform the test, using a solution of magnesium nitratehexahydrate in ethanol (95) (1 in 5) (not more than 1 ppm).

(3) Related substance 1—Weigh accurately about 50 mgof Cefotiam Hexetil Hydrochloride, and dissolve in a mix-ture of diluted phosphoric acid (1 in 100) and acetonitrile(4:1) to make exactly 50 mL. Pipet 10 mL of this solution,add a mixture of diluted phosphoric acid (1 in 100) andacetonitrile (4:1) to make exactly 25 mL, and use this solu-tion as the sample solution. Separately, weigh accuratelyabout 50 mg of Cefotiam Hexetil Hydrochloride ReferenceStandard, and dissolve in a mixture of diluted phosphoricacid (1 in 100) and acetonitrile (4:1) to make exactly 50 mL.Pipet 1 mL of this solution, add a mixture of diluted phos-phoric acid (1 in 100) and acetonitrile (4:1) to make exactly50 mL, and use this solution as the standard solution.Perform the test with exactly 10 mL each of the sample solu-tion and the standard solution as directed under the LiquidChromatography according to the following conditions, anddetermine each peak area by the automatic integrationmethod. Calculate the amount of the related substances bythe following equation: the amount of the related substancehaving the relative retention time of about 1.2 to one of thepeaks of cefotiam hexetil, which has the larger retentiontime, is not more than 2.0z, and each amount of the otherrelated substances is not more than 0.5z. For this calcula-tion, use the value of the peak area obtained by the automat-ic integration method of the related substance having the rel-ative retention time of about 1.2 to one of the peaks ofcefotiam hexetil, which has the larger retention time, aftermultiplying by its sensitivity coe‹cient, 0.78.

Amount (z) of each related substance＝ WS

WT×

AT

AS× 5

WS: Amount (g) of Cefotiam Hexetil HydrochlorideReference Standard

WT: Amount (g) of the sampleAS: Total of two peak areas of cefotiam hexetil from the

standard solutionAT: Each peak area of related substance from the sample

solution





Mobile phase A: A mixture of diluted 0.2 mol/L potassi-


um dihydrogen phosphate TS (1 in 2), acetonitrile and aceticacid (100) (72:28:1).

Mobile phase B: A mixture of acetonitrile, diluted0.2 mol/L potassium dihydrogen phosphate TS (1 in 2) andacetic acid (100) (60:40:1).

Flowing of the mobile phase: Adjust so that the mixingrate of the mobile phase A and the mobile phase B ischanged lineally from 1:0 to 0:1 for 30 minutes.

Flow rate: 0.7 mL per minute.Time span of measurement: As long as about 3 times of

the retention time of one of the cefotiam hexetil peaks,which appears ˆrst, after the solvent peak.System suitability—

Test for required detectability: Measure exactly 1 mL ofthe standard solution, and add a mixture of diluted phos-phoric acid (1 in 100) and acetonitrile (4:1) to make exactly50 mL. Conˆrm that each area of the two peaks of cefotiamhexetil obtained from 10 mL of this solution is equivalent to1.6 to 2.4z of that from 10 mL of the standard solution.

System performance: When the procedure is run with10 mL of the standard solution under the above operatingconditions, the resolution between the two peaks of cefotiamhexetil is not less than 2.0.

System repeatability: When the test is repeated 6 timeswith 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the total ofthe two peak areas of cefotiam hexetil is not more than2.0z.

(4) Related substance 2—Weigh accurately about 20 mgof Cefotiam Hexetil Hydrochloride, dissolve in exactly 2 mLof methanol, add a mixture of a solution of diammoniumhydrogen phosphate (79 in 20,000) and acetic acid (100)(200:3) to make exactly 50 mL, and use this solution as thesample solution. Separately, weigh accurately about 20 mgof Cefotiam Hydrochloride Reference Standard, and dis-solve in the mobile phase to make exactly 50 mL. Pipet 2 mLof this solution, add the mobile phase to make exactly50 mL, and use this solution as the standard solution. Per-form the test with exactly 10 mL each of the sample solutionand the standard solution as directed under the Liquid Chro-matography according to the following conditions, anddetermine each peak area by the automatic integrationmethod. Calculate the amount of the related substances bythe following equation: the amounts of the related sub-stances having the relative retention time of about 0.1 and0.9 to cefotiam are not more than 1.0z, respectively, andeach amount of the related substances other than the relatedsubstances having the relative retention time of about 0.1and 0.9 to cefotiam is not more than 0.5z. For this calcula-tion, use the value of the peak area of the related substancehaving the relative retention time of about 0.9 to cefotiamafter multiplying by its sensitivity coe‹cient, 0.76.

Amount (z) of each related substance＝WS

WT×

AT

AS× 4

WS: Amount (g) of Cefotiam Hydrochloride ReferenceStandard

WT: Amount (g) of the sample

AS: Peak area of cefotiam from the standard solutionAT: Each peak area from the sample solution





Mobile phase: A mixture of a solution of diammoniumhydrogen phosphate (79 in 20,000), methanol and acetic acid(100) (200:10:3).

Flow rate: Adjust the ‰ow rate so that the retention timeof cefotiam is about 15 minutes.

Time span of measurement: As long as about 2 times ofthe retention time of cefotiam after the solvent peak.System suitability—

Test for required detectability: Measure exactly 1 mL ofthe standard solution, and add the mobile phase to makeexactly 50 mL. Conˆrm that the peak area of cefotiamobtained from 10 mL of this solution is equivalent to 1.6 to2.4z of that from 10 mL of the standard solution.

System performance: To 1 mL of a solution ofacetaminophen in the mobile phase (1 in 50,000) add 3 mLof the standard solution, and mix well. When the procedureis run with 10 mL of this solution under the above operatingconditions, acetaminophen and cefotiam are eluted in thisorder with the resolution between these peaks being not lessthan 4.

System repeatability: When the test is repeated 6 timeswith 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peakarea of cefotiam is not more than 2.0z.

(5) Total amount of related substances—The total of theamount of related substances obtained in the Related sub-stance 1 and the Related substance 2 is not more than 6.5z.



Isomer ratio Proceed the test with 20 mL of the samplesolution obtained in the Assay as directed under the LiquidChromatography according to the conditions directed in theAssay, and determine the areas of the two peaks, Aa for thefaster peak and Ab for the later peak, closely appeared eachother at the retention time of around 10 minutes: Aa/(Aa＋

Ab) is not less than 0.45 and not more than 0.55.

Assay Weigh accurately an amount of Cefotiam HexetilHydrochloride and Cefotiam Hexetil Hydrochloride Refer-ence Standard, equivalent to about 30 mg (potency), anddissolve each in a mixture of diluted phosphoric acid (1 in100) and acetonitrile (4:1) to make exactly 50 mL. Measureexactly 5 mL each of these solutions, add exactly 5 mL of theinternal standard solution and a mixture of diluted phos-phoric acid (1 in 100) and acetonitrile (4:1) to make exactly


50 mL, and use these solutions as the sample solution andthe standard solution. Perform the test with 20 mL each ofthe sample solution and the standard solution as directedunder the Liquid Chromatography according to the follow-ing conditions, and determine the ratios, QT and QS, of thepeak area of cefotiam hexetil to that of the internal stand-ard. For this calculation, the total of the areas of the twopeaks appeared closely each other at the retention time ofaround 10 minutes is used as the peak area of cefotiamhexetil.

Amount [mg (potency)] of cefotiam (C18H23N9O4S3)

＝WS×QT

QS× 1000

WS: Amount [mg (potency)] of Cefotiam Hexetil Hydro-chloride Reference Standard

Internal standard solution—A solution of benzoic acid in amixture of diluted phosphoric acid (1 in 100) and acetonitrile(4:1) (7 in 10,000).Operating conditions—




Mobile phase: A mixture of diluted 0.2 mol/L potassiumdihydrogen phosphate TS (1 in 2), acetonitrile and aceticacid (100) (72:28:1).

Flow rate: Adjust the ‰ow rate so that the retention timeof the faster peak of cefotiam hexetil is about 9 minutes.System suitability—

System performance: When the procedure is run with20 mL of the standard solution under the above operatingconditions, the internal standard and cefotiam hexetil areeluted in this order with the resolution between the twopeaks of cefotiam hexetil being not less than 2.0.

System repeatability: When the test is repeated 6 timeswith 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratiosof the peak area of cefotiam hexetil to that of the internalstandard is not more than 1.0z.


Cefoxitin Sodiumセフォキシチンナトリウム


Cefoxitin Sodium contains not less than 927 mg(potency) and not more than 970 mg (potency) per mg,calculated on the dehydrated, de-acetone and de-methanol basis. The potency of Cefoxitin Sodiumis expressed as mass (potency) of cefoxitin(C16H17N3O7S2: 427.45).

Description Cefoxitin Sodium occurs as white to light yel-lowish white granules or powder.

It is very soluble in water, soluble in methanol, and slight-ly soluble in ethanol (95).

Identiˆcation (1) Determine the absorption spectrum ofa solution of Cefoxitin Sodium in 0.05 mol/L phosphatebuŠer solution, pH 7.0 (1 in 40,000) as directed under theUltraviolet-visible Spectrophotometry, and compare thespectrum with the Reference Spectrum: both spectra exhibitsimilar intensities of absorption at the same wavelengths.

(2) Determine the spectrum of a solution of CefoxitinSodium in heavy water for nuclear magnetic resonance spec-troscopy (1 in 10) as directed under the Nuclear MagneticResonance Spectroscopy (1H), using sodium 3-trimethylsilyl-propanesulfonate for nuclear magnetic resonance spec-troscopy as an internal reference compound: it exhibitssingle signals, A, B and C, at around d3.5 ppm, at aroundd3.9 ppm and at around d5.1 ppm, respectively, and a mul-tiple signal D between d6.9 ppm and d7.5 ppm. The ratio ofthe integrated intensity of each signal, A:B:C:D, is about3:2:1:3.

(3) A solution of Cefoxitin Sodium (1 in 10) responds tothe Qualitative Tests for sodium salt.


the dehydrated, de-acetone and de-methanol basis,methanol, 25 mL, 100 mm).

pH The pH of a solution obtained by dissolving 1.0 g ofCefoxitin Sodium in 10 mL of water is between 4.5 and 6.5.

Purity (1) Clarity and color of solution—A solutionobtained by dissolving 1.0 g of Cefoxitin Sodium in 10 mLof water is clear and a pale yellow to yellow.

(2) Heavy metals—Proceed with 1.0 g of CefoxitinSodium according to Method 2, and perform the test. Pre-pare the control solution with 2.0 mL of Standard LeadSolution (not more than 20 ppm).

(3) Arsenic—Prepare the test solution with 1.0 g ofCefoxitin Sodium according to Method 3, and perform thetest (not more than 2 ppm).

(4) Acetone and methanol—Dissolve 5.0 g of CefoxitinSodium in water to make exactly 50 mL. Pipet 3 mL of thissolution in a centrifuge tube, allow to stand in an ice-coldwater for 2 minutes, and add exactly 3 mL of diluted


hydrochloric acid (1 in 50). After mixing, centrifuge, and usethe supernatant liquid as the sample solution. Separately, toexactly 5 mL of acetone add water to make exactly 1000 mL,and use this solution as the standard stock solution (1).Separately, to exactly 5 mL of methanol add water to makeexactly 1000 mL, and use this solution as the standard stocksolution (2). To exactly 50 mL of the standard stock solution(1) and exactly 5 mL of the standard stock solution (2) addwater to make exactly 500 mL, and use this solution as thestandard solution. Perform the test with exactly 2 mL each ofthe sample solution and the standard solution as directed un-der the Gas Chromatography according to the followingconditions, and determine the peak areas of acetone, ATa

and ASa, and the peak areas of methanol, ATb and ASb, ofthese solutions: the amounts of acetone and methanol arenot more than 0.7z and not more than 0.1z, respectively.

Amount (z) of acetone＝ ATa

ASa× 0.791

Amount (z) of methanol＝ATb

ASb× 0.0791

Operating conditions—Detector: An hydrogen ‰ame-ionization detector.Column: A glass column 3 mm in inside diameter and 2 m

in length, packed with porous styrene-divinylbenzenecopolymer for gas chromatography (150 – 180 mm in particlediameter, 0.0085 mm in average pore size, 300 – 400 m2/g).



of acetone is about 7 minutes.System suitability—

System performance: When the procedure is run with 2 mLof the standard solution under the above operating condi-tions, methanol and acetone are eluted in this order, and thenumber of theoretical plates of these peaks are not less than160, respectively, and the symmetry constant of these peaksare not more than 2.3 and not more than 1.3, respectively.

System repeatability: When the test is repeated 5 timeswith 2 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peakarea of acetone is not more than 5z.

(5) Lactone substance—Dissolve 0.25 g of CefoxitinSodium in 10 mL of water, and use this solution as the sam-ple solution. Pipet 0.5 mL of the sample solution, add waterto make exactly 100 mL, and use this solution as the stan-dard solution. Perform the test with these solutions asdirected under the Thin-layer Chromatography. Spot 5 mLeach of the sample solution and the standard solution on aplate of silica gel with ‰uorescent indicator for thin-layerchromatography, develop with a mixture of ethyl acetate,acetone, water and acetic acid (100) (5:2:1:1) to a distance ofabout 15 cm, and air-dry the plate. Examine under ultravio-let light (main wavelength: 254 nm): the spot other than theprincipal spot from the sample solution is not more intensethan the principal spot from the standard solution.


Assay Weigh accurately an amount of Cefoxitin Sodiumand Cefoxitin Reference Standard, equivalent to about 0.2 g(potency), and dissolve each in phosphate buŠer solution,pH 6.0 to make exactly 100 mL. Pipet 15 mL each of thesesolutions, add exactly 5 mL of the internal standard solutionand phosphate buŠer solution, pH 6.0 to make 50 mL, anduse these solutions as the sample solution and the standardsolution, respectively. Perform the test with 10 mL each ofthe sample solution and the standard solution as directedunder the Liquid Chromatography according to the follow-ing conditions, and determine the ratios, QT and QS, of thepeak area of cefoxitin to that of the internal standard.

Amount [mg (potency)] of cefoxitin (C16H17N3O7S2)

＝WS×QT

QS× 1000

WS: Amount [mg (potency)] of Cefoxitin ReferenceStandard

Internal standard solution—A solution of anhydrouscaŠeine in phosphate buŠer solution, pH 6.0 (1 in 200).Operating conditions—





Flow rate: Adjust the ‰ow rate so that the retention timeof cefoxitin is about 7 minutes.System suitability—

System performance: When the procedure is run with10 mL of the standard solution under the above operatingconditions, the internal standard and cefoxitin are eluted inthis order with the resolution between these peaks being notless than 2.5.

System repeatability: When the test is repeated 5 timeswith 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratiosof the peak areas of cefoxitin to that of the internal standardis not more than 2.0z.



Cefpiramide Sodiumセフピラミドナトリウム


Cefpiramide Sodium contains not less than 900 mg(potency) per mg, calculated on the anhydrous basis.The potency of Cefpiramide Sodium is expressedas mass (potency) of cefpiramide (C25H24N8O7S2:612.64).

Description Cefpiramide Sodium occurs as white to yel-lowish white powder.

It is very soluble in dimethylsulfoxide, freely soluble inwater, sparingly soluble in methanol, and slightly soluble inethanol (95).

Identiˆcation (1) Determine the absorption spectrum ofa solution of Cefpiramide Sodium in 0.05 mol/L phosphatebuŠer solution, pH 7.0 (1 in 50,000) as directed under theUltraviolet-visible Spectrophotometry, and compare thespectrum with the Reference Spectrum: both spectra exhibitsimilar intensities of absorption at the same wavelengths.

(2) Determine the spectrum of a solution of CefpiramideSodium in deuterated dimethylsulfoxide for nuclear magnet-ic resonance spectroscopy (1 in 10) as directed under theNuclear Magnetic Resonance Spectroscopy (1H), using tetra-methylsilane for nuclear magnetic resonance spectroscopy asan internal reference compound: it exhibits single signals, A,B and C, at around d2.3 ppm, at around d3.9 ppm and ataround d8.2 ppm, respectively. The ratio of the integratedintensity of each signal, A:B:C, is about 3:3:1.

(3) Cefpiramide Sodium responds to the Qualitative Test(1) for sodium salt.


anhydrous basis, 0.05 mol/L phosphate buŠer solution, pH7.0, 10 mL, 100 mm).

pH The pH of a solution obtained by dissolving 2.0 g ofCefpiramide Sodium in 20 mL of water is between 5.5 and8.0.

Purity (1) Clarity and color of solution—Dissolve 1.0 gof Cefpiramide Sodium in 10 mL of 0.05 mol/L phosphatebuŠer solution, pH 7.0: the solution is clear, and colorless orlight yellow.

(2) Heavy metals—Proceed with 1.0 g of CefpiramideSodium according to Method 2, and perform the test.Prepare the control solution with 2.0 mL of Standard LeadSolution (not more than 20 ppm).

(3) Related substances—Weigh accurately about 25 mgof Cefpiramide Sodium, dissolve in 0.03 mol/L phosphatebuŠer solution, pH 7.5 to make exactly 50 mL, and use thissolution as the sample solution. Separately, weigh accuratelyabout 25 mg of 1-methyl-1H-tetrazole-5-thiol for liquidchromatography, previously dried in a desiccator (in vacu-um, silica gel) for 2 hours, and an amount of Cefpiramide

Reference Standard, equivalent to about 75 mg (potency),dissolve them in 0.03 mol/L phosphate buŠer solution, pH7.5 to make exactly 100 mL. Pipet 2 mL of this solution, add0.03 mol/L phosphate buŠer solution, pH 7.5 to make ex-actly 100 mL, and use this solution as the standard solution.Perform the test with exactly 5 mL of the sample solutionand the standard solution as directed under the Liquid Chro-matography according to the following conditions, and de-termine each peak area by the automatic integrationmethod. Calculate the amount of 1-methyl-1H-tetrazole-5-thiol, each of the other related substances and the total ofthe other related substances by the following equations: theamount of 1-methyl-1H-tetrazole-5-thiol, each of the otherrelated substances and the total of the other related sub-stances are not more than 1.0z, not more than 1.5z andnot more than 4.0z, respectively.

Amount (z) of 1-methyl-1H-tetrazole-5-thiol (C2H4N4S)

＝WSa

WT×

ATa

ASa

Amount (z) of each of other related substances

＝WSb

WT×

ATc

ASb

WSa: Amount (mg) of 1-methyl-1H-tetrazole-5-thiolWSb: Amount [mg (potency)] of Cefpiramide Reference

StandardWT: Amount (mg) of the sampleASa: Peak area of 1-methyl-1H-tetrazole-5-thiol from the

standard solutionASb: Peak area of cefpiramide from the standard solutionATa: Peak area of 1-methyl-1H-tetrazole-5-thiol from the

sample solutionATc: Area of each peak other than 1-methyl-1H-tetrazole-

5-thiol and cefpiramide from the sample solution



and 30 cm in length, packed with octylsilanized silica gel forliquid chromatography (10 mm in particle diameter).


Mobile phase: A mixture of 0.03 mol/L phosphate buŠersolution, pH 7.5 and methanol (3:1).

Flow rate: Adjust the ‰ow rate so that the retention timeof cefpiramide is about 11 minutes.

Time span of measurement: About 2 times as long as theretention time of cefpiramide.System suitability—

Test for required detectability: Measure exactly 5 mL ofthe standard solution, and add 0.03 mol/L phosphate buŠersolution, pH 7.5 to make exactly 50 mL. Conˆrm that thepeak area of 1-methyl-1H-tetrazole-5-thiol obtained from5 mL of this solution is equivalent to 8 to 12z of that from5 mL of the standard solution.

System performance: Dissolve 25 mg of CefpiramideReference Standard and 7 mg of cinnamic acid in the mobile


phase to make 50 mL. When the procedure is run with 5 mLof this solution under the above operating conditions, cin-namic acid and cefpiramide are eluted in this order with theresolution between these peaks being not less than 3.

System repeatability: When the test is repeated 6 timeswith 5 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peakarea of 1-methyl-1H-tetrazole-5-thiol is not more than2.0z.


Assay Weigh accurately an amount of CefpiramideSodium and Cefpiramide Reference Standard, equivalent toabout 50 mg (potency), add exactly 5 mL of the internalstandard solution to dissolve, then add the mobile phase tomake 100 mL, and use these solutions as the sample solutionand the standard solution. Perform the test with 5 mL eachof the sample solution and the standard solution as directedunder the Liquid Chromatography according to the follow-ing conditions, and determine the ratios, QT and QS, of thepeak area of cefpiramide to that of the internal standard.

Amount [mg (potency)] of cefpiramide (C25H24N8O7S2)

＝WS×QT

QS× 1000

WS: Amount [mg (potency)] of Cefpiramide ReferenceStandard

Internal standard solution—A solution of 4-dimethylamino-antipyrine (1 in 100).Operating conditions—


Column: A stainless steel column 4 mm in inside diameterand 30 cm in length, packed with octylsilanized silica gel forliquid chromatography (10 mm in particle diameter).


Mobile phase: A mixture of 0.01 mol/L phosphatebuŠer solution, pH 6.8, acetonitrile, methanol and tetra-hydrofuran (22:1:1:1).

Flow rate: Adjust the ‰ow rate so that the retention timeof cefpiramide is about 7 minutes.System suitability—

System performance: When the procedure is run with 5 mLof the standard solution under the above operating condi-tions, cefpiramide and the internal standard are eluted inthis order with the resolution between these peaks being notless than 7.

System repeatability: When the test is repeated 6 timeswith 5 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio ofthe peak area of cefpiramide to that of the internal standardis not more than 2.0z.

Containers and storage Containers—Tight containers.Storage—Light-resistant, and at a temperature not ex-

ceeding 59C.

Add the following:

Cefpodoxime Proxetilセフポドキシムプロキセチル

C21H27N5O9S2: 557.60(1RS )-1-(Isopropoxycarbonyloxy)ethyl (6R,7R )-7-[(Z )-2-(2-aminothiazol-4-yl)-2-(methoxyimino)acetylamino]-3-methoxymethyl-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylate [87239-81-4]

Cefpodoxime Proxetil contains not less than 706 mg(potency) per mg, calculated on the anhydrous basis.The potency of Cefpodoxime Proxetil is expressedas mass (potency) of cefpodoxime (C15H17N5O6S2:427.46).

Description Cefpodoxime Proxetil occurs as a white tolight brownish white powder.

It is very soluble in acetonitrile, in methanol and in chlo-roform, freely soluble in ethanol (99.5), and very slightlysoluble in water.

Identiˆcation (1) Determine the absorption spectrum ofa solution of Cefpodoxime Proxetil in acetonitrile (3 in200,000) as directed under the Ultraviolet-visible Spec-trophotometry, and compare the spectrum with the Refer-ence Spectrum or the spectrum of a solution of Cef-podoxime Proxetil Reference Standard prepared in the samemanner as the sample solution: both spectra exhibit similarintensities of absorption at the same wavelengths.

(2) Determine the infrared absorption spectrum of Cef-podoxime Proxetil as directed in the potassium bromide diskmethod under the Infrared Spectrophotometry, and com-pare the spectrum with the Reference Spectrum or the spec-trum of Cefpodoxime Proxetil Reference Standard: bothspectra exhibit similar intensities of absorption at the samewave numbers.

(3) Determine the spectrum of a solution of Cef-podoxime Proxetil in deuterochloroform for nuclearmagnetic resonance spectroscopy (1 in 10) as directed underthe Nuclear Magnetic Resonance Spectroscopy (1H), usingtetramethylsilane for nuclear magnetic resonance spec-troscopy as an internal reference compound: it exhibits dou-ble signals, A and B, at around d1.3 ppm and at aroundd1.6 ppm, and single signals, C and D, at around d3.3 ppmand at around d4.0 ppm. The ratio of the integrated inten-sity of these signals, A:B:C:D, is about 2:1:1:1.

Optical rotation [a]20D : ＋24.0 –＋31.49(0.1 g calculated on

the anhydrous basis, acetonitrile, 20 mL, 100 mm).


Purity (1) Heavy metals—Proceed with 1.0 g of Cef-podoxime Proxetil according to Method 2, and perform thetest. Prepare the control solution with 2.0 mL of StandardLead Solution (not more than 20 ppm).

(2) Related substances—Dissolve 50 mg of CefpodoximeProxetil in 50 mL of a mixture of water, acetonitrile andacetic acid (100) (99:99:2), and use this solution as the sam-ple solution. Perform the test with 20 mL of the sample solu-tion as directed under the Liquid Chromatography accord-ing to the following conditions. If necessary, perform thetest in the same manner with 20 mL of the mixture of water,acetonitrile and acetic acid (100) (99:99:2) to compensate forthe base line. Determine each peak area by the automaticintegration method, and calculate the amounts of them bythe area percentage method: the peak, having the relativeretention time of about 0.8 with respect to the isomer B ofcefpodoxime proxetil, is not more than 2.0z, the peak otherthan cefpodoxime proxetil is not more than 1.0z, and thesum of the peaks other than cefpodoxime proxetil is notmore than 6.0z.Operating conditions—




Mobile phase A: A mixture of water, methanol and a solu-tion of formic acid (1 in 50) (11:8:1).

Mobile phase B: A mixture of methanol and a solution offormic acid (1 in 50) (19:1).



Mobile phaseA (z)

Mobile phaseB (z)

0 – 65 95 565 – 145 95→ 15 5→ 85

145 – 155 15 85

Flow rate: Adjust the ‰ow rate so that the retention timeof the isomer B of cefpodoxime proxetil is about 60 minutes.

Time span of measurement: About 2.5 times as long as theretention time of the isomer B of cefpodoxime proxetil afterthe solvent peak.System suitability—

Test for required detectability: Measure exactly 5 mL ofthe sample solution, add the mixture of water, acetonitrileand acetic acid (100) (99:99:2) to make exactly 200 mL, anduse this solution as the solution for required detectabilitytest. Pipet 2 mL of the solution for required detectabilitytest, and add the mixture of water, acetonitrile and aceticacid (100) (99:99:2) to make exactly 100 mL. Conˆrm thatthe peak areas of the isomer A and the isomer B of cef-podoxime proxetil obtained from 20 mL of this solution are

equivalent to 1.4 to 2.6z of them from 20 mL of the solutionfor required detectability test, respectively.

System performance: Dissolve 1 mg of CefpodoximeProxetil in 100 mL of the mixture of water, acetonitrile andacetic acid (100) (99:99:2). When the procedure is run with20 mL of this solution under the above operating conditions,the isomer A and the isomer B of cefpodoxime proxetil areeluted in this order with the resolution between these peaksbeing not less than 6.

System repeatability: Dissolve 1 mg of CefpodoximeProxetil in 100 mL of the mixture of water, acetonitrile andacetic acid (100) (99:99:2). When the test is repeated 5 timeswith 20 mL of this solution under the above operating condi-tions, the relative standard deviations of the peak areas ofthe isomer A and the isomer B are not more than 2.0z,respectively.



Isomer ratio Perform the test with 5 mL of the sample solu-tion obtained in the Assay as directed under the LiquidChromatography according to the following conditions, anddetermine the peak areas, Aa and Ab, of the two isomers ofcefpodoxime proxetil, having the smaller and larger reten-tion times, respectively, by the automatic integrationmethod: Ab/(Aa＋ Ab) is between 0.50 and 0.60.Operating conditions—

Detector, column, column temperature, mobile phase,and ‰ow rate: Proceed as directed in the operating condi-tions in the Assay.System suitability—


Assay Weigh accurately an amount of Cefpodoxime Prox-etil and Cefpodoxime Proxetil Reference Standard, equiva-lent to about 60 mg (potency), dissolve in 80 mL of acetoni-trile, add exactly 4 mL of the internal standard solution, addacetonitrile to make 100 mL, and use these solutions as thesample solution and the standard solution. Perform the testwith 5 mL each of the sample solution and the standard solu-tion as directed under the Liquid Chromatography accord-ing to the following conditions, and determine the ratios,QT1, QS1, QT2 and QS2, of the areas of the two peaks of theisomers of cefpodoxime proxetil to the peak area of the in-ternal standard.

Amount [mg (potency)] of cefpodoxime (C15H17N5O6S2)

＝WS×QT1＋ QT2

QS1＋ QS2× 1000

WS: Amount [mg (potency)] of Cefpodoxime ProxetilReference Standard

Internal standard solution—Dissolve 0.3 g of ethyl para-hydroxybenzoate in a solution of citric acid in acetonitrile (1in 2000) to make 100 mL.






Mobile phase: A mixture of water and methanol (11:9).Flow rate: Adjust the ‰ow rate so that the retention time

of the internal standard is about 11 minutes.System suitability—

System performance: When the procedure is run with 5 mLof the standard solution under the above operating condi-tions, the internal standard, the isomer A and the isomer Bare eluted in this order with the resolution between thesepeaks being not less than 4.

System repeatability: When the test is repeated 5 timeswith 5 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratiosof the peak area of the isomer B of cefpodoxime proxetil tothat of the internal standard is not more than 1.0z.


Cefroxadineセフロキサジン


Cefroxadine contains not less than 921 mg (potency)per mg, calculated on the dehydrated basis. Thepotency of Cefroxadine is expressed as mass (potency)of cefroxadine (C16H19N3O5S: 365.41).

Description Cefroxadine occurs as pale yellowish white tolight yellow, crystalline particles or powder.

It is very soluble in formic acid, slightly soluble in waterand in methanol, and very slightly soluble in acetonitrileand in ethanol (95).

It dissolves in 0.001 mol/L hydrochloric acid TS and indilute acetic acid.

Identiˆcation (1) Determine the absorption spectrum ofa solution of Cefroxadine in 0.001 mol/L hydrochloric acidTS (1 in 50,000) as directed under the Ultraviolet-visibleSpectrophotometry, and compare the spectrum with theReference Spectrum or the spectrum of a solution of Cefrox-adine Reference Standard prepared in the same manner asthe sample solution: both spectra exhibit similar intensitiesof absorption at the same wavelengths.

(2) Determine the spectrum of a solution of Cefroxadinein deuterated formic acid for nuclear magnetic resonancespectroscopy (1 in 10) as directed under the Nuclear Magnet-ic Resonance Spectroscopy (1H), using tetramethylsilane for

nuclear magnetic resonance spectroscopy as an internalreference compound: it exhibits three sharp single signals,A, B and C, at around d2.8 ppm, at around d4.1 ppm and ataround d6.3 ppm. The ratio of the integrated intensity ofeach signal, A:B:C, is about 4:3:1.

Optical rotation [a]20D :＋95 –＋1089(0.1 g calculated on the

dehydrated basis, diluted acetic acid (100) (3 in 25), 100 mL,100 mm).

Purity (1) Heavy metals—Weigh 1.0 g of Cefroxadine ina porcelain crucible, add 10 mL of a solution of magnesiumnitrate hexahydrate in ethanol (95) (1 in 10), mix, burn theethanol, and carbonize by gently heating. After cooling, add2 mL of nitric acid, heat carefully, and incinerate by ignitionat 500 – 6009C. If a carbonized substance is still remained,moisten it with a small amount of nitric acid, and incinerateagain by ignition. After cooling, add 6 mL of hydrochloricacid, and evaporate on a water bath to dryness. Moisten theresidue with 3 drops of hydrochloric acid, and add 10 mL ofhot water to dissolve the residue by heating on a water bath.After cooling, adjust the pH between 3 and 4 with ammoniaTS, add 2 mL of dilute acetic acid, ˆlter if necessary, trans-fer to a Nessler tube, wash the crucible with 10 mL of water,and add the washing and water to the tube to make 50 mL.Perform the test with this solution. Prepare the control solu-tion as follows: Put 2.0 mL of Standard Lead Solution and10 mL of a solution of magnesium nitrate hexahydrate inethanol (95) (1 in 10) in a porcelain crucible, and proceed asdirected for the preparation of the test solution (not morethan 20 ppm).

(2) Arsenic—Prepare the test solution with 1.0 g ofCefroxadine according to Method 4, and perform the test(not more than 2 ppm).

(3) Related substances—Dissolve 10 mg of Cefroxadinein 100 mL of the mobile phase, and use this solution as thesample solution. Pipet 1 mL of the sample solution, add themobile phase to make exactly 100 mL, and use this solutionas the standard solution. Perform the test with exactly 40 mLeach of the sample solution and the standard solution asdirected under the Liquid Chromatography according to thefollowing conditions, and determine each peak area ob-tained from the chromatograms of these solutions by the au-tomatic integration method: the areas of the peaks appearedat the relative retention times of 0.07, 0.6 and 0.8 against thepeak of cefroxadine from the sample solution are not morethan 2 times, 4 times and 1 time of the peak area of cefroxa-dine from the standard solution, respectively, and any peakarea other than cefroxadine and other than the peaks men-tioned above from the sample solution is not more than 1/2of the peak area of cefroxadine from the standard solution,and the total area of the peaks other than cefroxadine fromthe sample solution is not more than 6 times of the peak areaof cefroxadine from the standard solution.Operating conditions—


Column: A stainless steel column 4.6 mm in inside di-ameter and 10 cm in length, packed with octadecylsilanized


silica gel for liquid chromatography (5 mm in particle di-ameter).


Mobile phase: Dissolve 1.4 g of sodium perchlorate in1000 mL of a mixture of water and acetonitrile (489:11).

Flow rate: Adjust the ‰ow rate so that the retention timeof cefroxadine is about 20 minutes.

Time span of measurement: About 2 times as long as theretention time of cefroxadine.System suitability—

Test for required detectability: Measure exactly 2 mL ofthe standard solution, and add the mobile phase to makeexactly 20 mL. Conˆrm that the peak area of cefroxadineobtained from 40 mL of this solution is equivalent to 7 to13z of that obtained from 40 mL of the standard solution.

System performance: Dissolve 3 mg of Cefroxadine and15 mg of orcin in 100 mL of the mobile phase. When theprocedure is run with 40 mL of this solution under the aboveoperating conditions, orcin and cefroxadine are eluted inthis order with the resolution between these peaks being notless than 3.

System repeatability: When the test is repeated 6 timeswith 40 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peakarea of cefroxadine is not more than 2.0z.

Water Not less than 8.5z and not more than 12.0z

(0.1 g, volumetric titration, direct titration).

Assay Weigh accurately an amount of Cefroxadine andCefroxadine Reference Standard, equivalent to about 50 mg(potency), dissolve each in a suitable amount of a mixture ofdilute acetic acid and phosphoric acid (500:1), add exactly5 mL of the internal standard solution and a mixture ofdilute acetic acid and phosphoric acid (500:1) to make200 mL, and use these solutions as the sample solution andthe standard solution. Perform the test with 10 mL each ofthe sample solution and the standard solution as directedunder the Liquid Chromatography according to the follow-ing conditions, and determine the ratios, QT and QS, of thepeak area of cefroxadine to that of the internal standard.


＝WS×QT

QS× 1000

WS: Amount [mg (potency)] of Cefroxadine ReferenceStandard

Internal standard solution—Dissolve 1.6 g of vanillin in5 mL of methanol, and add a mixture of dilute acetic acidand phosphoric acid (500:1) to make 100 mL.Operating conditions—




Mobile phase: A mixture of a solution of ammonium sul-fate (1 in 50) and acetonitrile (97:3).

Flow rate: Adjust the ‰ow rate so that the retention timeof cefroxadine is about 10 minutes.System suitability—

System performance: When the procedure is run with10 mL of the standard solution under the above operatingconditions, cefroxadine and the internal standard are elutedin this order with the resolution between these peaks beingnot less than 1.5.

System repeatability: When the test is repeated 6 timeswith 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratiosof the peak areas of cefroxadine to that of the internal stan-dard is not more than 1.0z.


Cefteram Pivoxilセフテラムピボキシル


Cefteram Pivoxil contains not less than 743 mg(potency) per mg, calculated on the anhydrous basis.The potency of Cefteram Pivoxil is expressed as mass(potency) of cefteram (C16H17N9O5S2: 479.49).

Description Cefteram Pivoxil occurs as a white to paleyellowish white powder.

It is very soluble in acetonitrile, freely soluble inmethanol, in ethanol (95) and in chloroform, and practicallyinsoluble in water.

Identiˆcation (1) Determine the absorption spectrum ofa solution of Cefteram Pivoxil in 0.05 mol/L hydrochloricacid-methanol TS (1 in 100,000) as directed under theUltraviolet-visible Spectrophotometry, and compare thespectrum with the Reference Spectrum: both spectra exhibitsimilar intensities of absorption at the same wavelengths.

(2) Determine the spectrum of a solution of CefteramPivoxil in deuterated chloroform for nuclear magneticresonance spectroscopy (1 in 10) as directed under theNuclear Magnetic Resonance Spectroscopy (1H), usingtetramethylsilane for nuclear magnetic resonance spec-troscopy as an internal reference compound: it exhibitssingle signals A, B and C, at around d1.2 ppm, at aroundd2.5 ppm and at around d4.0 ppm, respectively. The ratioof the integrated intensity of these signals, A:B:C, is about3:1:1.


anhydrous basis, methanol, 20 mL, 100 mm).

Purity (1) Heavy metals—Proceed with 1.0 g of Cefter-


am Pivoxil according to Method 2, and perform the test.Prepare the control solution with 2.0 mL of Standard LeadSolution (not more than 20 ppm).

(2) Arsenic—Prepare the test solution with 1.0 g ofCefteram Pivoxil according to Method 4, and perform thetest (not more than 2 ppm).

(3) Related substances—Dissolve 50 mg of CefteramPivoxilin in 50 mL of the mobile phase, and use this solutionas the sample solution. Pipet 1 mL of the sample solution,add the mobile phase to make exactly 50 mL, and use thissolution as the standard solution. Perform the test with ex-actly 10 mL each of the sample solution and the standard so-lution as directed under the Liquid Chromatography accord-ing to the following conditions, and determine each peakarea by the automatic integration method: the area of thepeak, having the relative retention time of about 0.9 withrespect to cefteram pivoxil from the sample solution is notmore than 1.25 times the peak area of cefteram pivoxil fromthe standard solution, the area of the peak, having the rela-tive retention time of about 0.1 with respect to cefterampivoxil from the sample solution is not more than 0.25 timesthe peak area of cefteram pivoxil from the standard solu-tion, and the total area of the peaks other than cefterampivoxil from the sample solution is not more than 2.75 timesthe peak area of cefteram pivoxil from the standard solu-tion. For the above calculation, use the area of the peak,having the relative retention time of about 0.1, after mul-tiplying by its sensitivity coe‹cient, 0.74.Operating conditions—


Time span of measurement: About 2 times as long as theretention time of cefteram pivoxil.System suitability—

Test for required detectability: Measure exactly 1 mL ofthe standard solution, and add the mobile phase to makeexactly 10 mL. Conˆrm that the peak area of cefterampivoxil obtained from 10 mL of this solution is equivalent to7 to 13z of that from 10 mL of the standard solution.

System performance: When the procedure is run with10 mL of the standard solution under the above operatingconditions, the number of theoretical plates and the symmet-ry coe‹cient of the peak of cefteram pivoxil are not less than5000 and not more than 1.5, respectively.

System repeatability: When the test is repeated 6 timeswith 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peakarea of cefteram pivoxil is not more than 3.0z.

Water Not more than 3.0z (0.3 g, coulometric titration).

Assay Weigh accurately an amount of Cefteram Pivoxiland Cefteram Pivoxil Mesitylenesulfonate Reference Stan-dard, equivalent to about 40 mg (potency), dissolve each in20 mL of diluted acetonitrile (1 in 2), add exactly 5 mL ofthe internal standard solution and diluted acetonitrile (1 in 2)to make 50 mL, and use these solutions as the sample solu-tion and the standard solution. Perform the test with 10 mL

each of the sample solution and the standard solution asdirected under the Liquid Chromatography according to thefollowing conditions, and determine the ratios, QT and QS,of the peak area of cefteram pivoxil to that of the internalstandard.

Amount [mg (potency)] of cefteram (C16H17N9O5S2)

＝WS×QT

QS× 1000

WS: Amount [mg (potency)] of Cefteram Pivoxil Mesity-lenesulfonate Reference Standard

Internal standard solution—A solution of methyl para-hydroxybenzoate in diluted acetonitrile (1 in 2) (1 in 1000).Operating conditions—




Mobile phase: To 100 mL of acetic acid-sodium acetatebuŠer solution, pH 5.0 add 375 mL of acetonitrile and waterto make 1000 mL.

Flow rate: Adjust the ‰ow rate so that the retention timeof cefteram pivoxil is about 14 minutes.System suitability—

System performance: When the procedure is run with10 mL of the standard solution under the above operatingconditions, the internal standard and cefteram pivoxil areeluted in this order with the resolution between these peaksbeing not less than 3.

System repeatability: When the test is repeated 6 timeswith 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratiosof the peak area of cefteram pivoxil to that of the internalstandard is not more than 1.0z.

Containers and storage Containers—Tight containers.Storage—In a cold place.

Ceftibutenセフチブテン

Change the Identiˆcation (1) to read:

Identiˆcation (1) Determine the absorption spectrum ofa solution of Ceftibuten in 0.1 mol/L phosphate buŠer solu-tion for antibiotics, pH 8.0 (1 in 50,000) as directed underthe Ultraviolet-visible Spectrophotometry: it exhibits a max-imum between 261 nm and 265 nm.

Change the Absorbance to read:

Absorbance E 1z1 cm (263 nm): 320 – 345 (20 mg calculated on

the anhydrous basis, 0.1 mol/L phosphate buŠer solution


for antibiotics, pH 8.0, 1000 mL).

Change the Optical rotation to read:


the anhydrous basis, 0.1 mol/L phosphate buŠer solutionfor antibiotics, pH 8.0, 50 mL, 100 mm).


Assay Weigh accurately an amount of Ceftibuten andCeftibuten Hydrochloride Reference Standard, equivalentto about 10 mg (potency), dissolve each in 36 mL of0.1 mol/L phosphate buŠer solution for antibiotics, pH 8.0,add exactly 4 mL each of the internal standard solution,shake, and use these solutions as the sample solution and thestandard solution. Perform the test with 5 mL of the samplesolution and the standard solution as directed under the Liq-uid Chromatography according to the following conditions,and calculate the ratios, QT and QS, of the peak area ofceftibuten to that of the internal standard. Keep the samplesolution and the standard solution at 59C or below and usewithin 2 hours.


＝WS×QT

QS× 1000

WS: Amount [mg (potency)] of Ceftibuten HydrochlorideReference Standard

Internal standard solution—A solution of methyl p-hydrox-ybenzoate in acetonitrile (3 in 4000).Operating conditions—




Mobile phase: A mixture of 0.005 mol/L n-decyltrimethylammonium bromide TS and acetonitrile (4:1).

Flow rate: Adjust the ‰ow rate so that the retention timeof ceftibuten is about 10 minutes.System suitability—

System performance: Dissolve 5 mg of Ceftibuten in1 mol/L Hydrochloric acid TS to make 50 mL, and allow tostand for 4 hours at room temperature. To 10 mL of thissolution add 0.1 mol/L phosphate buŠer solution forantibiotics, pH 8.0 to make 25 mL. When the procedure isrun with 5 mL of this solution under the above operatingconditions, trans-isomer and ceftibuten are eluted in thisorder with the resolution between these peaks being not lessthan 1.5.

System repeatability: When the test is repeated 6 timeswith 5 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratiosof the peak area of ceftibuten to that of the internal standardis not more than 1.0z.

Ceftriaxone Sodiumセフトリアキソンナトリウム


Identiˆcation (1) Determine the absorption spectrum ofa solution of Ceftriaxone Sodium (1 in 100,000) as directedunder the Ultraviolet-visible Spectrophotometry, and com-pare the spectrum with the Reference Spectrum or the spec-trum of a solution of Ceftriaxone Sodium Reference Stan-dard prepared in the same manner as the sample solution:both spectra exhibit similar intensities of absorption at thesame wavelengths.


Purity(4) Related substances 1—Dissolve 20 mg of Ceftriaxone

Sodium in 10 mL of the mobile phase, and use this solutionas the sample solution. Pipet 1 mL of the sample solution,add a mixture of water and acetonitrile for liquid chro-matography (11:9) to make exactly 100 mL, and use this so-lution as the standard solution. Perform the test with exactly10 mL each of the sample solution and the standard solutionas directed under the Liquid Chromatography according tothe following conditions, and determine each peak area bythe automatic integration method: the peak areas of theimpurity 1 having the relative retention time of about 0.5and the impurity 2 having the relative retention time ofabout 1.3 to ceftriaxone from the sample solution are notmore than the peak area of ceftriaxone from the standardsolution. In this case, these peak areas for the impurity 1 andthe impurity 2 are used after multiplying by 0.9 and 1.2,respectively.Operating conditions—




Mobile phase: Dissolve 5.796 g of anhydrous disodiumhydrogen phosphate and 3.522 g of potassium dihydrogenphosphate in water to make exactly 1000 mL, and use thissolution as the solution A. Separately, dissolve 20.256 g ofcitric acid monohydrate and 7.840 g of sodium hydroxide inwater to make exactly 1000 mL, and use this solution as thesolution B. Dissolve 4.00 g of tetra-n-heptylammoniumbromide in 450 mL of acetonitrile for liquid chromato-graphy, add 55 mL of the solution A, 5 mL of the solution Band 490 mL of water.

Flow rate: Adjust the ‰ow rate so that the retention timeof ceftriaxone is about 7 minutes.

Time span of measurement: About 2 times as long as theretention time of ceftriaxone.


System suitability—Test for required detectability: Measure exactly 5 mL of

the sample solution, add a mixture of water and acetonitrilefor liquid chromatography (11:9) to make exactly 200 mL,and use this solution as the solution for system suitabilitytest. Pipet 1 mL of the solution for system suitability test,and add a mixture of water and acetonitrile for liquid chro-matography (11:9) to make exactly 100 mL. Conˆrm thatthe peak area of ceftriaxone obtained from 10 mL of thissolution is equivalent to 0.9 to 1.1z of that from 10 mL ofthe solution for system suitability test.

System performance: Dissolve 10 mg of CeftriaxoneSodium in a mixture of water and acetonitrile for liquidchromatography (11:9) to make 5 mL, add 5 mL of a solu-tion of diethyl terephthalate in a mixture of water andacetonitrile for liquid chromatography (11:9) (9 in 5000),and add a mixture of water and acetonitrile for liquid chro-matography (11:9) to make 200 mL. When the procedure isrun with 10 mL of this solution under the above operatingconditions, ceftriaxone and diethyl terephthalate are elutedin this order, with the resolution between these peaks beingnot less than 6.

System repeatability: When the test is repeated 6 timeswith 10 mL of the solution for system suitability test underthe above operating conditions, the relative standard devia-tion of the peak area of ceftriaxone is not more than 1.0z.

(5) Related substances 2—Dissolve 10 mg of CeftriaxoneSodium in 10 mL of the mobile phase, and use this solutionas the sample solution. Pipet 1 mL of the sample solution,add a mixture of acetonitrile for liquid chromatography andwater (23:11) to make exactly 100 mL, and use this solutionas the standard solution. Perform the test with exactly 10 mLeach of the sample solution and the standard solution asdirected under the Liquid Chromatography according to thefollowing conditions, and determine each peak area by theautomatic integration method: the each peak area of the im-purities which appear after the peak of ceftriaxone from thesample solution is not more than the peak area ofceftriaxone from the standard solution, and the total peakarea of these impurities is not more than 2.5 times of thepeak area from the standard solution.Operating conditions—




Mobile phase: Dissolve 5.796 g of anhydrous disodiumhydrogen phosphate and 3.522 g of potassium dihydrogenphosphate in water to make exactly 1000 mL, and use thissolution as the solution A. Separately, dissolve 20.256 g ofcitric acid monohydrate and 7.840 g of sodium hydroxide inwater to make exactly 1000 mL, and use this solution as thesolution B. Dissolve 4.00 g of tetra-n-heptylammoniumbromide in 450 mL of acetonitrile for liquid chro-

matography, and add 55 mL of the solution A, 5 mL of thesolution B, 490 mL of water and 700 mL of acetonitrile forliquid chromatography.

Flow rate: Adjust the ‰ow rate so that the retention timeof ceftriaxone is about 3 minutes.

Time span of measurement: About 10 times as long as theretention time of ceftriaxone.System suitability—

Test for required detectability: Measure exactly 5 mL ofthe sample solution, add a mixture of acetonitrile for liquidchromatography and water (23:11) to make exactly 100 mL,and use this solution as the solution for system suitabilitytest. Measure exactly 1 mL of the solution for systemsuitability test, and add a mixture of acetonitrile for liquidchromatography and water (23:11) to make exactly 100 mL.Conˆrm that the peak area of ceftriaxone obtained from10 mL of this solution is equivalent to 0.9 to 1.1z of thatfrom 10 mL of the solution for system suitability test.

System performance: Dissolve 10 mg of CeftriaxoneSodium in a mixture of acetonitrile for liquid chro-matography and water (23:11) to make 5 mL, add 5 mL of asolution of diethyl terephthalate in a mixture of water andacetonitrile for liquid chromatography (11:9) (9 in 5000),and add a mixture of acetonitrile for liquid chromatographyand water (23:11) to make 200 mL. When the procedure isrun with 10 mL of this solution under the above operatingconditions, ceftriaxone and diethyl terephthalate are elutedin this order with the resolution between these peaks beingnot less than 3.

System repeatability: When the test is repeated 6 timeswith 10 mL of the solution for system suitability test underthe above operating conditions, the relative standard devia-tion of the peak area of ceftriaxone is not more than 1.0z.


Assay Weigh accurately an amount of Ceftriaxone Sodiumand Ceftriaxone Sodium Reference Standard, equivalent toabout 0.1 g (potency), dissolve each in a mixture of waterand acetonitrile for liquid chromatography (11:9) to makeexactly 50 mL. Pipet 5 mL of each solution, add exactly5 mL of the internal standard solution and a mixture ofwater and acetonitrile for liquid chromatography (11:9) tomake 200 mL, and use these solutions as the sample solutionand the standard solution, respectively. Perform the testwith 10 mL each of these solutions as directed under the Liq-uid Chromatography according to the following conditions,and calculate the ratios, QT and QS, of the peak area ofceftriaxone to that of the internal standard.

Amount [mg (potency)] of ceftriaxone (C18H18N8O7S3)

＝WS×QT

QS× 1000

WS: Amount [mg (potency)] of Ceftriaxone Sodium Ref-erence Standard

Internal standard solution—A solution of diethyl terephtha-late in a mixture of water and acetonitrile for liquid chro-matography (11:9) (9 in 5000).






Mobile phase: Dissolve 5.796 g of anhydrous disodiumhydrogen phosphate and 3.522 g of potassium dihydrogenphosphate in water to make exactly 1000 mL, and use thissolution as solution A. Dissolve 20.256 g of citric acidmonohydrate and 7.840 g of sodium hydroxide in water tomake exactly 1000 mL, and use this solution as solution B.Dissolve 4.00 g of tetra-n-heptylammonium bromide in450 mL of acetonitrile for liquid chromatography, and add490 mL of water, 55 mL of solution A, and 5 mL of solutionB.

Flow rate: Adjust the ‰ow rate so that the retention timeof ceftriaxone is about 7 minutes.System suitability—

System performance: When the procedure is run with10 mL of the standard solution under the above operatingconditions, ceftriaxone and the internal standard are elutedin this order with the resolution between these peaks beingnot less than 6.

System repeatability: When the test is repeated 6 timeswith 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratiosof the peak area of ceftriaxone to that of the internal stan-dard is not more than 1.0z.

Cefuroxime Axetilセフロキシムアキセチル


Cefuroxime Axetil contains not less than 760 mg(potency) per mg, calculated on the anhydrous basisand corrected by the amount of acetone. The potencyof Cefuroxime Axetil is expressed as mass (potency) ofcefuroxime (C16H16N4O8S: 424.39).

Description Cefuroxime Axetil occurs as white to yellow-ish white, non-crystalline powder.

It is freely soluble in dimethylsulfoxide, soluble inmethanol, sparingly soluble in ethanol (95), and very slightlysoluble in water.

Identiˆcation (1) Determine the absorption spectrum ofa solution of Cefuroxime Axetil in methanol (3 in 200,000)as directed under the Ultraviolet-visible Spectrophotometry,and compare the spectrum with the Reference Spectrum orthe spectrum of a solution of Cefuroxime Axetil ReferenceStandard prepared in the same manner as the sample solu-

tion: both spectra exhibit similar intensities of absorption atthe same wavelengths.

(2) Determine the infrared absorption spectrum ofCefuroxime Axetil as directed in the potassium bromide diskmethod under the Infrared Spectrophotometry, and com-pare the spectrum with the Reference Spectrum or the spec-trum of Cefuroxime Axetil Reference Standard: both spec-tra exhibit similar intensities of absorption at the same wavenumbers.

(3) Determine the spectrum of a solution of CefuroximeAxetil in deuterated dimethylsulfoxide for nuclear magneticresonance spectroscopy (1 in 20) as directed under theNuclear Magnetic Resonance Spectroscopy (1H), usingtetramethylsilane for nuclear magnetic resonance spec-troscopy as an internal reference compound: it exhibits adouble signal or a pair of double signals A at aroundd1.5 ppm, a pair of single signals B at around d2.1 ppm,and a single signal C at around d3.9 ppm. The ratio of theintegrated intensity of each signal, A:B:C, is about 1:1:1.

Optical rotation [a]20D : ＋41 –＋479 (0.5 g, methanol,

50 mL, 100 mm).

Purity (1) Heavy metals—Proceed with 2.0 g ofCefuroxime Axetil according to Method 2, and perform thetest. Prepare the control solution with 2.0 mL of StandardLead Solution (not more than 10 ppm).

(2) Arsenic—Put 1.0 g of Cefuroxime Axetil in a cruci-ble, add 10 mL of a solution of magnesium nitrate hexahy-drate in ethanol (95) (1 in 10), burn the ethanol, then heatgradually to incinerate. If a carbonized substance remains,moisten with a small amount of nitric acid, and ignite to in-cinerate. Cool, add 10 mL of dilute hydrochloric acid to theresidue, dissolve by warming on a water bath, and performthe test using this solution as the test solution (not more than2 ppm).

(3) Related substance—Dissolve 25 mg of CefuroximeAxetil in 4 mL of methanol, add a solution of ammoniumdihydrogen phosphate (23 in 1000) to make 10 mL, and usethis solution as the sample solution. Pipet 1 mL of thesample solution, add 40 mL of methanol and a solution ofammonium dihydrogen phosphate (23 in 1000) to make ex-actly 100 mL, and use this solution as the standard solution.Perform the test with exactly 2 mL each of the sample solu-tion and the standard solution as directed under the LiquidChromatography according to the following conditions, anddetermine each peak area by the automatic integrationmethod: the area of the peak other than cefuroxime axetilobtained from the sample solution is not more than 1.5 timesthe sum area of two peaks of cefuroxime axetil obtainedfrom the standard solution, and the sum area of the peaksother than cefuroxime axetil from the sample solution is notmore than 4 times the sum area of two peaks of cefuroximeaxetil from the standard solution.Operating conditions—


Time span of measurement: About 3 times as long as the


retention time of the peak having the larger retention time ofthe two peaks of cefuroxime axetil after the solvent peak.System suitability—

Test for required detectability: Measure exactly 1 mL ofthe standard solution, and add 4 mL of methanol and a solu-tion of ammonium dihydrogen phosphate (23 in 1000) tomake exactly 10 mL. Conˆrm that the sum area of the twopeaks of cefuroxime axetil obtained from 2 mL of this solu-tion is equivalent to 7 to 13z of that obtained from 2 mL ofthe standard solution.


System repeatability: When the test is repeated 6 timeswith 2 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the sumarea of the two peaks of cefuroxime axetil is not more than2.0z.

(4) Acetone—Weigh accurately about 1.0 g ofCefuroxime Axetil, add exactly 0.2 mL of the internal stan-dard solution and dimethylsulfoxide to make exactly 10 mL,and use this solution as the sample solution. Separately,weigh accurately about 0.5 g of acetone, and add dimethyl-sulfoxide to make exactly 100 mL. Pipet 0.2 mL of this solu-tion, add exactly 0.2 mL of the internal standard solutionand dimethylsulfoxide to make exactly 10 mL, and use thissolution as the standard solution. Perform the test with 1 mLeach of the sample solution and the standard solution asdirected under the Gas Chromatography according to thefollowing conditions, determine each peak area by theautomatic integration method, and calculate the ratios, QT

and QS, of the peak area of acetone to that of the internalstandard: the amount of acetone is not more than 1.3z.

Amount (z) of acetone＝WS

WT×

QT

QS× 0.2

WS: Amount (g) of acetoneWT: Amount (g) of the sample

Internal standard solution—A solution of 1-propanol indimethylsulfoxide (1 in 200).Operating conditions—

Detector: A hydrogen ‰ame-ionization detector.Column: A glass column 3 mm in inside diameter and 2 m

in length, packed with siliceous earth for gas chromato-graphy coated with a mixture of polyethylene glycol 600 forgas chromatography and polyethylene glycol 1500 for gaschromatography (1:1) in the ratio of 20z (125 – 150 mm inparticle diameter).


Temperature of injection port: A constant temperature ofabout 1159C.


of the internal standard is about 4 minutes.System suitability—

System performance: When the procedure is run with 1 mLof the standard solution under the above operating condi-

tions, acetone and the internal standard are eluted in thisorder with the resolution between these peaks being not lessthan 5.

System repeatability: When the test is repeated 6 timeswith 1 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratiosof the peak area of acetone to that of the internal standard isnot more than 5.0z.



Isomer ratio Perform the test with 10 mL of the sample so-lution obtained in the Assay as directed under the LiquidChromatography according to the following conditions, anddetermine the area, Aa, of the peak having the smaller reten-tion time and the area, Ab, of the peak having the biggerretention time of the two peaks of cefuroxime axetil:Ab/(Aa＋ Ab) is between 0.48 and 0.55.Operating conditions—



Assay Weigh accurately an amount of Cefuroxime Axetiland Cefuroxime Axetil Reference Standard, equivalent toabout 50 mg (potency), and dissolve each in methanol tomake exactly 50 mL. Pipet 10 mL each of these solutions,add exactly 5 mL of the internal standard solution, 5 mL ofmethanol and a solution of ammonium dihydrogen phos-phate (23 in 1000) to make 50 mL, and use these solutions asthe sample solution and the standard solution. Perform thetest with 10 mL each of the sample solution and the standardsolution as directed under the Liquid Chromatographyaccording to the following conditions, and determine theratios, QT and QS, of the sum area of the two peaks ofcefuroxime axetil to the peak area of the internal standard.

Amount [mg (potency)] of cefuroxime (C16H16N4O8S)

＝WS×QT

QS× 1000

WS: Amount [mg (potency)] of Cefuroxime Axetil Refer-ence Standard

Internal standard solution—A solution of acetanilide inmethanol (27 in 5000).Operating conditions—


Column: A stainless steel column 4.6 mm in inside di-ameter and 20 cm in length, packed with trimethylsilanizedsilica gel for liquid chromatography (5 mm in particle di-ameter).



Mobile phase: A mixture of a solution of ammoniumdihydrogen phosphate (23 in 1000) and methanol (5:3).

Flow rate: Adjust the ‰ow rate so that the retention timeof the peak having the smaller retention time of the twopeaks of cefuroxime axetil is about 8 minutes.System suitability—

System performance: When the procedure is run with10 mL of the standard solution under the above operatingconditions, the internal standard and cefuroxime axetil areeluted in this order with the resolution between the twopeaks of cefuroxime axetil being not less than 1.5.

System repeatability: When the test is repeated 6 timeswith 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratiosof the sum area of the two peaks of cefuroxime axetil to thepeak area of the internal standard is not more than 1.0z.


Cetraxate Hydrochloride塩酸セトラキサート

Change the Purity (3) and (4) to read:

Purity(3) cis Isomer—Dissolve 0.10 g of Cetraxate Hydrochlo-

ride in 10 mL of water, and use this solution as the samplesolution. To exactly 5 mL of the sample solution add waterto make exactly 100 mL. To exactly 2 mL of this solutionadd water to make exactly 50 mL, and use this solution asthe standard solution. Perform the test with exactly 10 mLeach of the sample solution and the standard solution asdirected under the Liquid Chromatography according to thefollowing conditions. Determine each peak area of both so-lutions by the automatic integration method: the area of thepeak which has a retention time 1.3 to 1.6 times that ofcetraxate from the sample solution is not larger than thepeak area of cetraxate from the standard solution.Operating conditions—




Mobile phase: Adjust the pH of a mixture of water,methanol and 0.5 mol/L ammonium acetate TS (15:10:4) to6.0 with acetic acid (31).

Flow rate: Adjust the ‰ow rate so that the retention timeof cetraxate is about 10 minutes.System suitability—

System performance: Dissolve 0.02 g of CetraxateHydrochloride and 0.01 g of phenol in 100 mL of water. To2 mL of this solution add water to make 20 mL. When theprocedure is run with 10 mL of this solution under the above

operating conditions, cetraxate and phenol are eluted in thisorder with the resolution between these peaks being not lessthan 5.

System repeatability: When the test is repeated 6 timeswith 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peakarea of cetraxate is not more than 2.0z.

(4) 3-(p-Hydroxyphenyl)propionic acid—To 0.10 g ofCetraxate Hydrochloride add exactly 2 mL of the internalstandard solution and methanol to make 10 mL, and use thissolution as the sample solution. Separately, dissolve 25 mgof 3-(p-hydroxyphenyl)propionic acid in methanol to makeexactly 100 mL. To exactly 2 mL of this solution add exactly2 mL of the internal standard solution and methanol tomake 10 mL, and use this solution as the standard solution.Perform the test with 10 mL each of the sample solution andthe standard solution as directed under the Liquid Chro-matography according to the following conditions, andcalculate the ratios, QT and QS, of the peak area of 3-(p-hydroxyphenyl)propionic acid to that of the internal stan-dard: QT is not larger than QS.Internal standard solution—A solution of caŠeine inmethanol (1 in 4000).Operating conditions—




Mobile phase: Adjust the pH of a mixture of water,methanol and 0.5 mol/L ammonium acetate TS (15:5:2) to5.5 with acetic acid (31).

Flow rate: Adjust the ‰ow rate so that the retention timeof 3-(p-hydroxyphenyl)propionic acid is about 7 minutes.System suitability—

System performance: When the procedure is run with10 mL of the standard solution under the above operatingconditions, 3-(p-hydroxyphenyl)propionic acid and the in-ternal standard are eluted in this order with the resolutionbetween these peaks being not less than 5.

System repeatability: When the test is repeated 6 timeswith 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratiosof the peak area of 3-(p-hydroxyphenyl)propionic acid tothat of the internal standard is not more than 1.0z.


Chloramphenicolクロラムフェニコール


Chloramphenicol contains not less than 980 mg(potency) per mg, calculated on the dried basis. Thepotency of Chloramphenicol is expressed as mass(potency) of chloramphenicol (C11H12Cl2N2O5).

Description Chloramphenicol occurs as white to yellowishwhite, crystals or crystalline powder.

It is freely soluble in methanol and in ethanol (99.5), andslightly soluble in water.

Identiˆcation (1) Determine the absorption spectrum ofthe sample solution obtained in the Assay as directed underthe Ultraviolet-visible Spectrophotometry, and compare thespectrum with the Reference Spectrum or the spectrum of asolution of Chloramphenicol Reference Standard preparedin the same manner as the sample solution: both spectraexhibit similar intensities of absorption at the same wave-lengths.

(2) Determine the infrared absorption spectrum ofChloramphenicol as directed in the potassium bromide diskmethod under the Infrared Spectrophotometry, and com-pare the spectrum with the Reference Spectrum or the spec-trum of Chloramphenicol Reference Standard: both spectraexhibit similar intensities of absorption at the same wavenumbers.

Optical rotation [a]20D : ＋18.5 –＋21.59 (1.25 g, ethanol

(99.5), 25 mL, 100 mm).

Melting point 150 – 1559C

Purity (1) Heavy metals—Proceed with 1.0 g of Chlo-ramphenicol according to Method 2, and perform the test.Prepare the control solution with 2.5 mL of Standard LeadSolution (not more than 25 ppm).

(2) Arsenic—Prepare the test solution with 2.0 g ofChloramphenicol according to Method 4, and perform thetest (not more than 1 ppm).

(3) Related substances—Dissolve 0.10 g of Chloram-phenicol in 10 mL of methanol, and use this solution as thesample solution. Pipet 1 mL of the sample solution, addmethanol to make exactly 100 mL, and use this solution asthe standard solution (1). Pipet 10 mL of the standard solu-tion (1), add methanol to make exactly 20 mL, and use thissolution as the standard solution (2). Perform the test withthese solutions as directed under the Thin-layer Chro-matography. Spot 20 mL each of the sample solution and thestandard solutions (1) and (2) on a plate of silica gel with‰uorescent indicator for thin-layer chromatography, de-velop the plate with a mixture of chloroform, methanol andacetic acid (100) (79:14:7) to a distance of about 15 cm, andair-dry the plate. Examine under ultraviolet light (mainwavelength: 254 nm): the spots other than the principal spot

and the spot on the original obtained from the sample solu-tion are not more intense than the spot from the standardsolution (1), and the total amount of these spots is not morethan 2.0z.



Assay Weigh accurately an amount of Chloramphenicoland Chloramphenicol Reference Standard, equivalent toabout 0.1 g (potency), dissolve each in 20 mL of methanol,and add water to make exactly 100 mL. Pipet 20 mL each ofthese solutions, and add water to make exactly 100 mL.Pipet 10 mL each of these solutions, add water to make ex-actly 100 mL, and use these solutions as the sample solutionand the standard solution. Determine the absorbances, AT

and AS, at 278 nm of the sample solution and the standardsolution as directed under the Ultraviolet-visible Spec-trophotometry.

Amount [mg (potency)] of C11H12Cl2N2O5

＝WS×AT

AS× 1000

WS: Amount [mg (potency)] of Chloramphenicol Refer-ence Standard


Add the following:

Chloramphenicol Palmitateパルミチン酸クロラムフェニコール

C27H42Cl2N2O6: 561.54(2R,3R )-2-(Dichloroacetyl)amino-3-hydroxy-3-(4-nitrophenyl)propan-1-yl palmitate [530-43-8]

Chloramphenicol Palmitate contains not less than558 mg (potency) per mg, calculated on the dried basis.The potency of Chloramphenicol Palmitate is ex-pressed as mass (potency) of chloramphenicol(C11H12Cl2N2O5: 323.13).

Description Chloramphenicol Palmitate occurs as a whiteto grayish white, crystalline powder.

It is freely soluble in acetone, sparingly soluble inmethanol and in ethanol (99.5), and practically insoluble inwater.

Identiˆcation (1) Determine the absorption spectrum ofa solution of Chloramphenicol Palmitate in ethanol (99.5) (1in 33,000) as directed under the Ultraviolet-visible Spec-


trophotometry, and compare the spectrum with the Refer-ence Spectrum or the spectrum of a solution of Chloram-phenicol Palmitate Reference Standard prepared in the samemanner as the sample solution: both spectra exhibit similarintensities of absorption at the same wavelengths.

(2) Dissolve 5 mg each of Chloramphenicol Palmitateand Chloramphenicol Palmitate Reference Standard in1 mL of acetone, and use these solutions as the sample solu-tion and the standard soution. Perform the test with thesesolutions as directed under the Thin-layer Chromatography.Spot 5 mL each of the sample solution and the standard solu-tion on a plate of silica gel with ‰uorescent indicator forthin-layer chromatography. Develop the plate with a mix-ture of acetone and cyclohexane (1:1) to a distance of about10 cm, and air-dry the plate. Examine under ultraviolet light(main wavelength: 254 nm): the principal spot obtainedfrom the sample solution has the same Rf value as the spotfrom the standard solution.


dried basis, ethanol (99.5), 20 mL, 100 mm).


Purity (1) Heavy metals—Proceed with 1.0 g of Chlo-ramphenicol Palmitate according to Method 4, and performthe test. Prepare the control solution with 2.0 mL of Stan-dard Lead Solution (not more than 20 ppm).

(2) Arsenic—Prepare the test solution with 1.0 g ofChloramphenicol Palmitate according to Method 3, and per-form the test (not more than 2 ppm).

(3) Related substances—Dissolve 50 mg of Chloram-phenicol Palmitate in 50 mL of methanol, and use this solu-tion as the sample solution. Pipet 1 mL of the sample solu-tion, add methanol to make exactly 100 mL, and use thissolution as the standard solution. Perform the test with ex-actly 20 mL each of the sample solution and the standard so-lution as directed under the Liquid Chromatography accord-ing to the following conditions. The test should be per-formed within 30 minutes after the sample solution and thestandard solution are prepared. Determine each peak areaby the automatic integration method: the total area of thepeaks other than the peak of chloramphenicol palmitatefrom the sample solution is not more than 3.5 times the peakarea of chloramphenicol palmitate from the standard solu-tion. For this calculation, use the peak areas for chloram-phenicol, having the relative retention time of about 0.5 withrespect to chloramphenicol palmitate, and for chloram-phenicol dipalmitate, having the relative retention time ofabout 5.0 with respect to chloramphenicol palmitate, aftermultiplying by their response factors, 0.5 and 1.4, respec-tively.Operating conditions—




Mobile phase: MethanolFlow rate: Adjust the ‰ow rate so that the retention time

of chloramphenicol palmitate is about 5 minutes.Time span of measurement: About 6 times as long as the

retention time of chloramphenicol palmitate.System suitability—

Test for required detectability: Dissolve 50 mg of Chlo-ramphenicol Palmitate in 50 mL of methanol. Pipet 1 mL ofthis solution, add methanol to make exactly 100 mL, and usethis solution as the solution for system suitability test. Pipet5 mL of the solution for system suitabillity test, and addmethanol to make exactly 50 mL. Conˆrm that the peak areaof chloramphenicol palmitate obtained from 20 mL of thissolution is equivalent to 7 to 13z of that obtained from20 mL of the solution for system suitability test.

System performance: When the procedure is run with20 mL of the solution for system suitability test under theabove operating conditions, the number of theoretical platesof the peak of chloramphenicol palmitate is not less than5000.

System repeatability: When the test is repeated 6 timeswith 20 mL of the solution for system suitability test underthe above operating conditions, the relative standard devia-tion of the peak area of chloramphenicol palmitate is notmore than 1.0z.


Assay Weigh accurately an amount of ChloramphenicolPalmitate and Chloramphenicol Palmitate Reference Stan-dard, equivalent to about 37 mg (potency), dissolve each in40 mL of methanol and exactly 1 mL of acetic acid (100),and add methanol to make exactly 50 mL. Pipet 10 mL eachof these solutions, add the mobile phase to make exactly25 mL, and use these solutions as the sample solution andthe standard solution. Perform the test with exactly 10 mLeach of the sample solution and the standard solution asdirected under the Liquid Chromatography according to thefollowing conditions, and determine the peak areas, AT andAS.

Amount [mg (potency)] of chloramphenicol (C11H12Cl2N2O5)

＝WS×AT

AS× 1000

WS: Amount [mg (potency)] of Chloramphenicol Palmi-tate Reference Standard






Mobile phase: A mixture of methanol, water and aceticacid (100) (172:27:1).

Flow rate: Adjust the ‰ow rate so that the retention timeof chloramphenicol palmitate is about 7 minutes.System suitability—

System performance: When the procedure is run with10 mL of the standard solution under the above operatingconditions, the number of theoretical plates of the peak ofchloramphenicol palmitate is not less than 2400.

System repeatability: When the test is repeated 6 timeswith 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peakarea of chloramphenicol palmitate is not more than 1.0z.


Add the following:

Chloramphenicol Sodium Succinateコハク酸クロラムフェニコールナトリウム

C15H15Cl2N2NaO8: 445.18Monosodium (2R,3R )-2-(dichloroacetyl)amino-3-hydroxy-3-(4-nitrophenyl)propan-1-yl succinate[982-57-0]

Chloramphenicol Sodium Succinate contains notless than 711 mg (potency) per mg, calculated on theanhydrous basis. The potency of ChloramphenicolSodium Succinate is expressed as mass (potency) ofchloramphenicol (C11H12Cl2N2O5: 323.13).

Description Chloramphenicol Sodium Succinate occurs aswhite to yellowish white, crystals or crystalline powder.

It is very soluble in water, and freely soluble in methanoland in ethanol (99.5).

It is hygroscopic.

Identiˆcation (1) Determine the absorption spectrum ofa solution of Chloramphenicol Sodium Succinate (1 in50,000) as directed under the Ultraviolet-visible Spec-trophotometry, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities ofabsorption at the same wavelengths.

(2) Determine the infrared absorption spectrum of Chlo-ramphenicol Sodium Succinate as directed in the potassiumbromide disk method under the Infrared Spectrophotomet-ry, and compare the spectrum with the Reference Spectrum:both spectra exhibit similar intensities of absorption at thesame wave numbers.

(3) Chloramphenicol Sodium Succinate responds to theQualitative Test (1) for sodium salt.



pH The pH of a solution obtained by dissolving 1.4 g ofChloramphenicol Sodium Succinate in 5 mL of water isbetween 6.0 and 7.0.

Purity (1) Clarity and color of solution—Dissolve 1.0 gof Chloramphenicol Sodium Succinate in 10 mL of water:the solution is clear and colorless to yellowish.

(2) Heavy metals—Proceed with 1.0 g of Chloram-phenicol Sodium Succinate according to Method 2, andperform the test. Prepare the control solution with 2.0 mLof Standard Lead Solution (not more than 20 ppm).

(3) Arsenic—Prepare the test solution with 1.0 g ofChloramphenicol Sodium Succinate according to Method 1,and perform the test (not more than 2 ppm).


Assay Weigh accurately an amount of ChloramphenicolSodium Succinate, equivalent to about 20 mg (potency), dis-solve in water to make exactly 1000 mL, and use this solu-tion as the sample solution. Separately, weigh accurately anamount of Chloramphenicol Succinate Reference Standard,equivalent to about 20 mg (potency), add about 50 mL ofwater to make a suspension, and add gradually about 7 mLof 0.01 mol/L sodium hydroxide TS while stirring to adjustthe pH to 7.0. To this solution add water to make exactly1000 mL, and use this solution as the standard solution.Determine the absorbances, AT and AS, at 276 nm of thesample solution and the standard solution as directed underthe Ultraviolet-visible Spectrophotometry.

Amount [mg (potency)] of chloramphenicol (C11H12Cl2N2O5)

＝WS×AT

AS× 1000

WS: Amount [mg (potency)] of Chloramphenicol Suc-cinate Reference Standard


Chlordiazepoxide Powderクロルジアゼポキシド散


Assay Conduct this procedure without exposure to day-light, using light-resistant vessels. Weigh accurately a quan-tity of Chlordiazepoxide Powder, equivalent to about 0.1 gof Chlordiazepoxide (C16H14ClN3O), transfer to a glass-stoppered ‰ask, wet with exactly 10 mL of water, add exact-ly 90 mL of methanol, stopper, shake vigorously for 15minutes, and centrifuge. Pipet 10 mL of the supernatant liq-uid, add exactly 5 mL of the internal standard solution, add


methanol to make exactly 100 mL, and use this solution asthe sample solution. Separately, weigh accurately about 0.1g of Chlordiazepoxide Reference Standard, previously driedin a desiccator (in vacuum, phosphorus (V) oxide, 609C) for4 hours, and dissolve in exactly 10 mL of water and 90 mLof methanol. Pipet 10 mL of this solution, add exactly 5 mLof the internal standard solution, add methanol to make 100mL, and use this solution as the standard solution. Performthe test with 10 mL each of the sample solution and the stan-dard solution as directed under the Liquid Chromatographyaccording to the following conditions, and calculate the ra-tios, QT and QS, of the peak area of chlordiazepoxide to thatof the internal standard.

Amount (mg) of chlordiazepoxide (C16H14ClN3O)

＝WS×QT

QS× 10

WS: Amount (mg) of Chlordiazepoxide ReferenceStandard

Internal standard solution—A solution of isobutyl salicylatein methanol (1 in 20).Operating conditions—




Mobile phase: A mixture of methanol and 0.02 mol/Lammonium dihydrogen phosphate TS (7:3).

Flow rate: Adjust the ‰ow rate so that the retention timeof chlordiazepoxide is about 5 minutes.System suitability—

System performance: When the procedure is run with10 mL of the standard solution under the above operatingconditions, chlordiazepoxide and the internal standard areeluted in this order with the resolution between these peaksbeing not less than 9.

System repeatability: When the test is repeated 6 timeswith 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratiosof the peak area of chlordiazepoxide to that of the internalstandard is not more than 1.0z.

Chlordiazepoxide Tabletsクロルジアゼポキシド錠


Assay Conduct this procedure without exposure to day-light, using light-resistant vessels. Weigh accurately a quan-tity of Chlordiazepoxide Tablets, equivalent to about 0.1 gof Chlordiazepoxide (C16H14ClN3O), transfer to a glass-stoppered ‰ask, add 10 mL of water, and shake well to disin-tegrate. Add 60 mL of methanol, shake well, add methanol

to make exactly 100 mL, and centrifuge. Pipet 10 mL of thesupernatant liquid, add exactly 5 mL of the internal stan-dard solution, add methanol to make exactly 100 mL, anduse this solution as the sample solution. Separately, weighaccurately about 10 mg of Chlordiazepoxide Reference Stan-dard, previously dried in a desiccator (in vacuum, phospho-rus (V) oxide, 609C) for 4 hours, dissolve in 1 mL of waterand a suitable amount of methanol, add exactly 5 mL of theinternal standard solution, add methanol to make 100 mL,and use this solution as the standard solution. Perform thetest with 10 mL each of the sample solution and the standardsolution as directed under the Liquid Chromatography ac-cording to the following conditions, and calculate the ratios,QT and QS, of the peak area of chlordiazepoxide to that ofthe internal standard.

Amount (mg) of chlordiazepoxide (C16H14ClN3O)

＝WS×QT

QS× 10

WS: Amount (mg) of Chlordiazepoxide ReferenceStandard

Internal standard solution—A solution of isobutyl salicylatein methanol (1 in 20).Operating conditions—




Mobile phase: A mixture of methanol and 0.02 mol/Lammonium dihydrogen phosphate TS (7:3).

Flow rate: Adjust the ‰ow rate so that the retention timeof chlordiazepoxide is about 5 minutes.System suitability—

System performance: When the procedure is run with10 mL of the standard solution under the above operatingconditions, chlordiazepoxide and the internal standard areeluted in this order with the resolution between these peaksbeing not less than 9.

System repeatability: When the test is repeated 6 timeswith 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratiosof the peak area of chlordiazepoxide to that of the internalstandard is not more than 1.0z.

Chlormadinone Acetate酢酸クロルマジノン


Purity(3) Other steroids—Dissolve 20 mg of Chlormadinone

Acetate in 10 mL of acetonitrile, and use this solution as thesample solution. Pipet 1 mL of the sample solution, add


acetonitrile to make exactly 100 mL, and use this solution asthe standard solution. Perform the test with exactly 10 mLeach of the sample solution and the standard solution asdirected under the Liquid Chromatography according to thefollowing conditions, and determine each peak area by theautomatic integration method: the total area of peaks otherthan the peak of chlormadinone acetate from the sample so-lution is not larger than the peak area of chlormadinoneacetate from the standard solution.Operating conditions—




Mobile phase: A mixture of acetonitrile and water (13:7).Flow rate: Adjust the ‰ow rate so that the retention time

of chlormadinone acetate is about 10 minutes.Time span of measurement: About 1.5 times as long as the

retention time of chlormadinone acetate after the solventpeak.System suitability—

Test for required detection: To exactly 5 mL of the stan-dard solution add acetonitorile to make exactly 50 mL. Con-ˆrm that the peak area of chlormadinone acetate obtainedfrom 10 mL of this solution is equivalent to 7 to 13z of thatof chlormadinone acetate obtained from 10 mL of the stan-dard solution.

System performance: Dissolve 8 mg of ChlormadinoneAcetate and 2 mg of butyl parahydroxybenzoate in 100 mLof acetonitrile. When the procedure is run with 10 mL of thissolution under the above operating conditions, butyl para-hydroxybenzoate and chlormadinone acetate are eluted inthis order with the resolution between these peaks being notless than 8.

System repeatability: When the test is repeated 6 timeswith 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peakarea of chlormadinone acetate is not more than 1.0z.

Chlorphenesin Carbamateカルバミン酸クロルフェネシン


Purity(3) Related substances—(i) Chlorphenesin-2-carba-

mate: Dissolve 0.10 g of Chlorphenesin Carbamate in 20 mLof a mixture of hexane for liquid chromatography and 2-propanol (7:3), and use this solution as the sample solution.Perform the test with 10 mL of the sample solution as direct-ed under the Liquid Chromatography according to thefollowing conditions. Determine the peak area, Aa, of chlor-phenesin carbamate and the peak area, Ab, of chlor-

phenesin-2-carbamate by the automatic integration method:the ratio, Ab/(Aa＋ Ab), is not larger than 0.007.Operating conditions—


Column: A stainless steel column 4 mm in inside diameterand 30 cm in length, packed with silica gel for liquid chro-matography (5 mm in particle diameter).


Mobile phase: A mixture of hexane for liquid chro-matography, 2-propanol and acetic acid (100) (700:300:1).

Flow rate: Adjust the ‰ow rate so that the retention timeof chlorphenesin carbamate is about 9 minutes.System suitability—

Test for required detection: Pipet 1 mL of the samplesolution, add a mixture of hexane for liquid chromato-graphy and 2-propanol (7:3) to make exactly 100 mL, anduse this solution as the solution for system suitability test.To exactly 5 mL of the solution for system suitability testadd a mixture of hexane for liquid chromatography and 2-propanol (7:3) to make exactly 10 mL. Conˆrm that thepeak area of chlorphenesin carbamate obtained from 10 mLof this solution is equivalent to 40 to 60z of that of chlor-phenesin carbamate obtained from 10 mL of the solution forsystem suitability test.

System performance: Dissolve 0.1 g of ChlorphenesinCarbamate in 50 mL of methanol. To 25 mL of this solutionadd 25 mL of dilute sodium hydroxide TS, and warm at609C for 20 minutes. To 20 mL of this solution add 5 mL of1 mol/L hydrochloric acid TS, shake well with 20 mL ofethyl acetate, and allow to stand to separate the ethyl acetatelayer. When the procedure is run with 10 mL of this layerunder the above operating conditions, chlorphenesin, chlor-phenesin carbamate and chlorphenesin-2-carbamate areeluted in this order, with the ratios of the retention time ofchlorphenesin and chlorphenesin-2-carbamate with respectto chlorphenesin carbamate are about 0.7 and about 1.2,respectively, and with the resolution between the peaks ofchlorphenesin and chlorphenesin carbamate being not lessthan 2.0.

System repeatability: When the test is repeated 6 timeswith 10 mL of the solution for system suitability test underthe above operating conditions, the relative standard devia-tion of the peak area of chlorphenesin carbamate is not morethan 2.0z.

(ii) Other related substances: Dissolve 0.10 g of Chlor-phenesin Carbamate in 10 mL of ethanol (95), and use thissolution as the sample solution. Pipet 1 mL of the samplesolution, add ethanol (95) to make exactly 20 mL. Pipet2 mL of this solution, add ethanol (95) to make exactly20 mL, and use this solution as the standard solution. Per-form the test with these solutions as directed under the Thin-layer Chromatography. Spot 50 mL each of the sample solu-tion and the standard solution on a plate of silica gel forthin-layer chromatography. Develop the plate with a mix-ture of ethyl acetate, methanol and ammonia solution (28)(17:2:1) to a distance of about 10 cm, and air-dry the plate.


Allow the plate to stand in iodine vapor for 20 minutes: thespots other than the principal spot from the sample solutionare not more intense than the spot from the standard solu-tion.

Ciclacillinシクラシリン


Ciclacillin contains not less than 920 mg (potency)per mg, calculated on the dehydrated basis. Thepotency of Ciclacillin is expressed as mass (potency) ofciclacillin (C15H23N3O4S).

Description Ciclacillin occurs as white to light yellowishwhite crystalline powder.

It is sparingly soluble in water, slightly soluble inmethanol, and practically insoluble in acetonitrile and inethanol (99.5).

Identiˆcation Determine the infrared absorption spectrumof Ciclacillin as directed in the potassium bromide diskmethod under the Infrared Spectrophotometry, and com-pare the spectrum with the Reference Spectrum or the spec-trum of Ciclacillin Reference Standard: both spectra exhibitsimilar intensities of absorption at the same wave numbers.

Optical rotation [a]20D : ＋300 –＋3159(2 g, water, 100 mL,

100 mm).

Purity (1) Heavy metals—Proceed with 1.0 g of Ciclacil-lin according to Method 2, and perform the test. Prepare thecontrol solution with 2.0 mL of Standard Lead Solution (notmore than 20 ppm).

(2) Arsenic—Prepare the test solution with 1.0 g ofCiclacillin according to Method 3, and perform the test (notmore than 2 ppm).


Assay Weigh accurately an amount of Ciclacillin andCiclacillin Reference Standard, equivalent to about 50 mg(potency), dissolve each in a suitable amount of the mobilephase, add exactly 5 mL of the internal standard solutionand the mobile phase to make 50 mL, and use these solutionsas the sample solution and the standard solution. Performthe test with 10 mL each of the sample solution and the stan-dard solution as directed under the Liquid Chromatographyaccording to the following conditions, and determine theratios, QT and QS, of the peak area of ciclacillin to that ofthe internal standard.


＝WS×QT

QS× 1000

WS: Amount [mg (potency)] of Ciclacillin Reference

Standard

Internal standard solution—A solution of orcin in themobile phase (1 in 500).Operating conditions—




Mobile phase: Dissolve 0.771 g of ammonium acetate inabout 900 mL of water, adjust the pH to 4.0 with acetic acid(100), and add water to make 1000 mL. To 850 mL of thissolution add 150 mL of acetonitrile.

Flow rate: Adjust the ‰ow rate so that the retention timeof ciclacillin is about 4 minutes.System suitability—

System performance: When the procedure is run with10 mL of the standard solution under the above operatingconditions, ciclacillin and the internal standard are eluted inthis order with the resolution between these peaks being notless than 8.

System repeatability: When the test is repeated 6 timeswith 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratiosof the peak areas of ciclacillin to that of the internal stan-dard is not more than 1.0z.


Citric Acidクエン酸


Citric Acid contains not less than 99.5z and notmore than 100.5z of anhydrous citric acid (C6H8O7:192.12), calculated on the anhydrous basis.

Description Citric Acid occurs as colorless crystals, whitegranules or crystalline powder.

It is very soluble in water, and freely soluble in ethanol(95).

It is eŒorescent in dry air.

Identiˆcation Determine the infrared absorption spectrumof Citric Acid, previously dried at 1059C for 2 hours, asdirected in the potassium bromide disk method under theInfrared Spectrophotometry, and compare the spectrumwith the Reference Spectrum: both spectra exhibit similarintensities of absorption at the same wave numbers.

Purity (1) Clarity and color of solution—Dissolve 2.0 gof Citric Acid in water to make 10 mL: the solution is clearand has no more color than the following control solutions


(1), (2) or (3).Control solution (1): To 1.5 mL of Cobalt (II) Chloride

Colorimetric Stock Solution and 6.0 mL of Iron (III)Chloride Colorimetric Stock Solution add water to make1000 mL.

Control solution (2): To 0.15 mL of Cobalt (II) ChlorideColorimetric Stock Solution, 7.2 mL of Iron (III) ChlorideColorimetric Stock Solution and 0.15 mL of Copper (II)Sulfate Colorimetric Stock Solution add water to make1000 mL.


(2) Sulfates—Dissolve 2.0 g of Citric Acid in water tomake 30 mL, and use this solution as the sample solution.Separately, dissolve 0.181 g of potassium sulfate in dilutedethanol (99.5) (3 in 10) to make exactly 500 mL. Pipet 5 mLof this solution, and add diluted ethanol (99.5) (3 in 10) tomake exactly 100 mL. To 4.5 mL of this solution add 3 mLof a solution of barium chloride dihydrate (1 in 4), shake,and allow to stand for 1 minute. To 2.5 mL of this solutionadd 15 mL of the sample solution and 0.5 mL of acetic acid(31), and allow to stand for 5 minutes: the solution has nomore turbidity than the following control solution.

Control solution: Dissolve 0.181 g of potassium sulfate inwater to make exactly 500 mL. Pipet 5 mL of this solution,add water to make exactly 100 mL, and proceed in the samemanner as above using this solution instead of the samplesolution.

(3) Oxalate—Dissolve 0.80 g of Citric Acid in 4 mL ofwater, add 3 mL of hydrochloric acid and 1 g of zinc, andboil for 1 minute. After allowing to stand for 2 minutes, takethe supernatant liquid, add 0.25 mL of a solution of phenyl-hydrazinium hydrochloride (1 in 100), heat to boil, and thencool quickly. To this solution add the equal volume ofhydrochloric acid and 0.25 mL of a solution of potassiumhexacyanoferrate (III) (1 in 20), mix, and allow to stand for30 minutes: the solution has no more color than the follow-ing control solution prepared at the same time.

Control solution: To 4 mL of a solution of oxalic aciddihydrate (1 in 10,000) add 3 mL of hydrochloric acid and1 g of zinc, and proceed in the same manner as the test solu-tion.

(4) Heavy metals—Proceed with 2.0 g of Citric Acidaccording to Method 2, and perform the test. Prepare thecontrol solution with 2.0 mL of Standard Lead Solution (notmore than 10 ppm).

(5) Readily carbonizable substances—Perform the testwith 0.5 g of Citric Acid, provided that the solution is heatedat 909C for 1 hour and then cool quickly: the solution has nomore color than Matching Fluid K.

Water Not less than 7.5z and not more than 9.0z (0.5 g,volumetric titration, direct titration).


Assay Weigh accurately about 0.55 g of Citric Acid,dissolve in 50 mL of water, and titrate with 1 mol/L sodiumhydroxide VS (indicator: 2 drops of phenolphthalein TS).

Each mL of 1 mol/L sodium hydroxide VS＝ 64.04 mg of C6H8O7


Anhydrous Citric Acid無水クエン酸


Anhydrous Citric Acid contains not less than 99.5zand not more than 100.5z of C6H8O7, calculated onthe anhydrous basis.

Description Anhydrous Citric Acid occurs as colorlesscrystals, white granules or crystalline powder.

It is very soluble in water, and freely soluble in ethanol(95).

Identiˆcation Determine the infrared absorption spectrumof Anhydrous Citric Acid, previously dried at 1059C for 24hours, as directed in the potassium bromide disk methodunder the Infrared Spectrophotometry, and compare thespectrum with the Reference Spectrum: both spectra exhibitsimilar intensities of absorption at the same wave numbers.

Purity (1) Clarity and color of solution—Dissolve 2.0 gof Anhydrous Citric Acid in water to make 10 mL: the solu-tion is clear and has no more color than the following con-trol solutions (1), (2) or (3).

Control solution (1): To 1.5 mL of Cobalt (II) ChlorideColorimetric Stock Solution and 6.0 mL of Iron (III)Chloride Colorimetric Stock Solution add water to make1000 mL.


Control solution (3): To 2.5 mL of Cobalt (II) ChlorideColorimetric Stock Solution, 6.0 mL of Iron (III) ChlorideColorimetric Stock Solution and 1.0 mL of Copper (II)Sulfate Colorimetric Stock Solution add water to make 1000mL.

(2) Sulfates—Dissolve 2.0 g of Anhydrous Citric Acid inwater to make 30 mL, and use this solution as the samplesolution. Separately, dissolve 0.181 g of potassium sulfate indiluted ethanol (99.5) (3 in 10) to make exactly 500 mL.Pipet 5 mL of this solution, and add diluted ethanol (99.5) (3in 10) to make exactly 100 mL. To 4.5 mL of this solutionadd 3 mL of a solution of barium chloride dihydrate (1 in 4),shake, and allow to stand for 1 minute. To 2.5 mL of this so-lution add 15 mL of the sample solution and 0.5 mL of acet-


ic acid (31), and allow to stand for 5 minutes: the solutionhas no more turbidity than the following control solution.

Control solution: Dissolve 0.181 g of potassium sulfate inwater to make exactly 500 mL. Pipet 5 mL of this solution,add water to make exactly 100 mL, and proceed in the samemanner as above using this solution instead of the samplesolution.

(3) Oxalate—Dissolve 0.80 g of Anhydrous Citric Acidin 4 mL of water, add 3 mL of hydrochloric acid and 1 g ofzinc, and boil for 1 minute. After allowing to stand for 2minutes, take the supernatant liquid, add 0.25 mL of a solu-tion of phenylhydrazinium hydrochloride (1 in 100), heat toboil, and then cool quickly. To this solution add the equalvolume of hydrochloric acid and 0.25 mL of a solution ofpotassium hexacyanoferrate (III) (1 in 20), mix, and allow tostand for 30 minutes: the solution has no more color thanthe following control solution prepared at the same time.

Control solution: To 4 mL of a solution of oxalic aciddihydrate (1 in 10,000) add 3 mL of hydrochloric acid and1 g of zinc, and proceed in the same manner as the test solu-tion.

(4) Heavy metals—Proceed with 2.0 g of AnhydrousCitric Acid according to Method 2, and perform the test.Prepare the control solution with 2.0 mL of Standard LeadSolution (not more than 10 ppm).

(5) Readily carbonizable substances—Perform the testwith 0.5 g of Anhydrous Citric Acid, provided that the solu-tion is heated at 909C for 1 hour and then cool quickly: thesolution has no more color than Matching Fluid K.



Assay Weigh accurately about 0.55 g of Anhydrous CitricAcid, dissolve in 50 mL of water, and titrate with 1 mol/Lsodium hydroxide VS (indicator: 2 drops of phenolphthaleinTS).

Each mL of 1 mol/L sodium hydroxide VS＝ 64.04 mg of C6H8O7


Add the following:

Clindamycin Hydrochloride塩酸クリンダマイシン

C18H33ClN2O5S.HCl: 461.44Methyl 7-chloro-6,7,8-trideoxy-6-[(2S,4R )-1-methyl-4-propylpyrrolidine-2-carboxamido]-1-thio-L-threo-a-D-galacto-octopyranoside monohydrochloride [21462-39-5]

Clindamycin Hydrochloride contains not less than759 mg (potency) per mg. The potency of ClindamycinHydrochloride is expressed as mass (potency) of clin-damycin (C18H33ClN2O5S: 424.98).

Description Clindamycin Hydrochloride occurs as white tograyish white, crystals or crystalline powder.

It is freely soluble in water and in methanol, and slightlysoluble in ethanol (95).

Identiˆcation Dissolve 0.1 g of Clindamycin Hydrochlo-ride in 5 mL of water, add 2 mL of sodium hydroxide TS,and mix: a white turbidity is produced. To this solution add0.3 mL of sodium pentacyanonitrosylferrate (III) TS, mix,allow to stand at 60 to 659C for 10 minutes, and add 2 mL ofdilute hydrochloric acid: a blue-green color develops.





(1) Test organism—Micrococcus luteus ATCC 9341(2) Culture medium—Use the medium i in 3) Medium

for other organisms under (1) Agar media for seed and baselayer.

(3) Standard solutions—Weigh accurately an amount ofClindamycin Hydrochloride Reference Standard, equivalentto about 25 mg (potency), dissolve in 0.1 mol/L phosphatebuŠer solution, pH 7.0 to make exactly 250 mL, and use thissolution as the standard stock solution. Keep the standardstock solution at a temperature not exceeding 159C and usewithin 14 days. Take exactly a suitable amount of the stan-dard stock solution before use, add 0.1 mol/L phosphatebuŠer solution, pH 7.0 to make solutions so that each mLcontains 2 mg (potency) and 1 mg (potency), and use thesesolutions as the high concentration standard solution andthe low concentration standard solution, respectively.

(4) Sample solutions—Weigh accurately an amount of


Clindamycin Hydrochloride, equivalent to about 25 mg(potency), and dissolve in 0.1 mol/L phosphate buŠer solu-tion, pH 7.0 to make exactly 250 mL. Take exactly a suitableamount of this solution, add 0.1 mol/L phosphate buŠersolution, pH 7.0 to make solutions so that each mL contains2 mg (potency) and 1 mg (potency), and use these solutions asthe high concentration sample solution and the low concen-tration sample solution, respectively.


Clindamycin Phosphateリン酸クリンダマイシン


Clindamycin Phosphate contains not less than758 mg (potency) per mg, calculated on the anhydrousbasis. The potency of Clindamycin Phosphate isexpressed as mass (potency) of clindamycin(C18H33ClN2O5S: 424.98).

Description Clindamycin Phosphate occurs as a white topale yellowish white crystalline powder.

It is freely soluble in water, sparingly soluble in methanol,and practically insoluble in ethanol (95).

Identiˆcation Determine the infrared absorption spectrumof Clindamycin Phosphate, previously dried at 1009C for 2hours, as directed in the paste method under the InfraredSpectrophotometry, and compare the spectrum with theReference Spectrum or the spectrum of Clindamycin Phos-phate Reference Standard previously dried at 1009C for 2hours: both spectra exhibit similar intensities of absorptionat the same wavelengths.



pH Dissolve 0.10 g of Clindamycin Phosphate in 10 mL ofwater. The pH of the solution is between 3.5 and 4.5.

Purity (1) Clarity and color of solution—Dissolve 1.0 gof Clindamycin Phosphate in 10 mL of freshly boiled andcooled water: the solution is clear and colorless.

(2) Heavy metals—Proceed with 2.0 g of ClindamycinPhosphate according to Method 4, and perform the test.Prepare the control solution with 1.0 mL of Standard LeadSolution (not more than 5 ppm).

(3) Arsenic—Prepare the test solution with 1.0 g of Clin-damycin Phosphate according to Method 4, and perform thetest (not more than 2 ppm).

(4) Related substances—Dissolve 0.1 g of ClindamycinPhosphate in 100 mL of the mobile phase, and use this solu-tion as the sample solution. Pipet 1 mL of the sample solu-tion, add the mobile phase to make exactly 100 mL, and usethis solution as the standard solution. Perform the test withexactly 20 mL each of the sample solution and the standard

solution as directed under the Liquid Chromatography ac-cording to the following conditions, and determine eachpeak area by the automatic integration method: the peakarea of clindamycin, having the relative retention time ofabout 1.8 with respect to clindamycin phosphate, obtainedfrom the sample solution is not more than 1/2 of the peakarea of clindamycin phosphate from the standard solution,and the total area of the peaks other than clindamycin phos-phate from the sample solution is not more than 4 times thepeak area of clindamycin phosphate from the standard solu-tion.Operating conditions—


Time span of measurement: About 2 times as long as theretention time of clindamycin phosphate after the solventpeak.System suitability—

Test for required detectability: Measure exactly 1 mL ofthe standard solution, and add the mobile phase to makeexactly 10 mL. Conˆrm that the peak area of clindamycinphosphate obtained from 20 mL of this solution is equivalentto 7 to 13z of that from 20 mL of the standard solution.



Assay Weigh accurately an amount of Clindamycin Phos-phate and Clindamycin Phosphate Reference Standard,equivalent to about 20 mg (potency), add exactly 25 mL ofthe internal standard solution and the mobile phase to make100 mL, and use these solutions as the sample solution andthe standard solution. Perform the test with 20 mL each ofthe sample solution and the standard solution as directed un-der the Liquid Chromatography according to the followingconditions, and determine the ratios, QT and QS, of the peakarea of clindamycin phosphate to that of the internal stan-dard.

Amount [mg (potency)] of clindamycin (C18H33ClN2O5S)

＝WS×QT

QS× 1000

WS: Amount [mg (potency)] of Clindamycin PhosphateReference Standard

Internal standard solution—A solution of methyl para-hydroxybenzoate in the mobile phase (3 in 50,000).Operating conditions—




Mobile phase: Dissolve 10.54 g of potassium dihydrogen


phosphate in 775 mL of water, adjust the pH to 2.5 withphosphoric acid, and add 225 mL of acetonitrile.

Flow rate: Adjust the ‰ow rate so that the retention timeof clindamycin phosphate is about 8 minutes.System suitability—

System performance: When the procedure is run with20 mL of the standard solution under the above operatingconditions, clindamycin phosphate and the internal standardare eluted in this order with the resolution between thesepeaks being not less than 4.

System repeatability: When the test is repeated 6 timeswith 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratiosof the peak area of clindamycin phosphate to that of theinternal standard is not more than 2.5z.


Clomifene Citrateクエン酸クロミフェン

Change the Isomer ratio to read:

Isomer ratio To 0.10 g of Clomifene Citrate add 10 mL ofwater and 1 mL of sodium hydroxide TS, and extract withthree 15-mL portions of diethyl ether. Wash the combineddiethyl ether extracts with 20 mL of water, add 10 g of anhy-drous sodium sulfate to the combined diethyl ether extracts,shake for 1 minute, ˆlter, and evaporate the diethyl ether ofthe ˆltrate. Dissolve the residue in 10 mL of chloroform, anduse this solution as the sample solution. Perform the testwith 2 mL of the sample solution as directed under the GasChromatography according to the following conditions.Determine the areas of two adjacent peaks, Aa and Ab, hav-ing retention times of about 20 minutes, where Aa is the peakarea of shorter retention time and Ab is the peak area oflonger retention time: AbW(Aa＋ Ab) is between 0.3 and 0.5.Operating conditions—

Detector: A hydrogen ‰ame-ionization detector.Column: A column 3 mm in inside diameter and 1 m in

length, having methylsilicone polymer coated at the ratioof 1z on siliceous earth for gas chromatography (125 to150 mm in particle diameter).



of the ˆrst peak of clomifene citrate is about 20 minutes.System suitability—

System performance: When the procedure is run with 2 mLof the sample solution under the above operating conditions,the resolution between the two peaks is not less than 1.3.

System repeatability: When the test is repeated 5 timeswith 2 mL of the sample solution under the above operatingconditions, the relative standard deviation of Ab/(Aa＋ Ab)is not more than 5.0z.

Cloxacillin Sodiumクロキサシリンナトリウム


Description Cloxacillin Sodium occurs as white to lightyellowish white, crystals or crystalline powder.

It is freely soluble in water, in N,N-dimethylformamideand in methanol, and sparingly soluble in ethanol (95).


Identiˆcation (1) Determine the absorption spectrum ofa solution of Cloxacillin Sodium in methanol (1 in 2500) asdirected under the Ultraviolet-visible Spectrophotometry,and compare the spectrum with the Reference Spectrum orthe spectrum of a solution of Cloxacillin Sodium ReferenceStandard prepared in the same manner as the sample solu-tion: both spectra exhibit similar intensities of absorption atthe same wavelength.




Change the pH to read:

pH Dissolve 1.0 g of Cloxacillin Sodium in 10 mL ofwater: the pH of the solution is between 6.0 and 7.5.


Purity (1) Clarity and color of solution—A solutionobtained by dissolving 1.0 g of Cloxacillin Sodium in 10 mLof water is clear and colorless to light yellow.

(2) Heavy metals—Proceed with 1.0 g of CloxacillinSodium according to Method 2, and perform the test. Pre-pare the control solution with 2.0 mL of Standard LeadSolution (not more than 20 ppm).

(3) Arsenic—Prepare the test solution with 1.0 g ofCloxacillin Sodium according to Method 5, and perform thetest (not more than 2 ppm).

(4) Related substances—Dissolve 50 mg of CloxacillinSodium in 50 mL of the mobile phase, and use this solutionas the sample solution. Pipet 1 mL of the sample solution,add the mobile phase to make exactly 100 mL, and use thissolution as the standard solution. Perform the test with ex-actly 10 mL each of the sample solution and the standard so-lution as directed under the Liquid Chromatography accord-ing to the following conditions, and determine each peakarea by the automatic integration method: the area of thepeak other than cloxacillin obtained from the sample solu-tion is not more than the peak area of cloxacillin obtainedfrom the standard solution.Operating conditions—


Column: A stainless steel column 6 mm in inside diameter




Mobile phase: Dissolve 4.953 g of diammonium hydrogenphosphate in 700 mL of water, and add 250 mL of acetoni-trile. Adjust the pH to 4.0 with phosphoric acid, and addwater to make exactly 1000 mL.

Flow rate: Adjust the ‰ow rate so that the retention timeof cloxacillin is about 24 minutes.

Time span of measurement: About 3 times as long as theretention time of cloxacillin.System suitability—

Test for required detectability: Measure exactly 1 mL ofthe standard solution, and add the mobile phase to makeexactly 10 mL. Conˆrm that the peak area of cloxacillinobtained from 10 mL of this solution is equivalent to 7 to13z of that obtained from the standard solution.

System performance: Weigh accurately about 50 mg ofCloxacillin Sodium Reference Standard, dissolve in a suita-ble amount of the mobile phase, add 5 mL of a solution ofguaifenesin in the mobile phase (1 in 200), then add themobile phase to make exactly 50 mL, and use this solution asthe solution for system suitability test. When the procedureis run with 10 mL of the solution for system suitability testunder the above operating conditions, guaifenesin and clox-acillin are eluted in this order with the resolution betweenthese peaks being not less than 25.

System repeatability: When the test is repeated 6 timeswith 10 mL of the solution for system suitability test underthe above operating conditions, the relative standard devia-tion of the ratios of the peak area of cloxacillin to that ofguaifenesin is not more than 1.0z.

Change the Water to read:

Water 3.0 – 4.5z (0.2 g, volumetric titration, direct titra-tion).

1z Codeine Phosphate Powderリン酸コデイン散 1


Assay Weigh accurately about 5 g of 1z Codeine Phos-phate Powder, dissolve in water to make exactly 100 mL,then pipet 10 mL of this solution, add exactly 10 mL of theinternal standard solution, and use this solution as the sam-ple solution. Separately, weigh accurately about 50 mg ofcodeine phosphate for assay, separately determined thewater in the same manner as Codeine Phosphate, dissolve inwater to make exactly 100 mL, then pipet 10 mL of this solu-tion, add exactly 10 mL of the internal standard solution,and use this solution as the standard solution. Perform thetest with 20 mL each of the sample solution and the standardsolution as directed under the Liquid Chromatography ac-cording to the following conditions, and calculate the ratios,

QT and QS, of the peak area of codeine to that of the inter-nal standard.

Amount (mg) of codeine phosphate(C18H21NO3.H3PO4.1/2H2O)

＝WS×QT

QS× 1.0227

WS: Amount (mg) of codeine phosphate for assay, calcu-lated on the anhydrous basis

Internal standard solution—A solution of etilefrinehydrochloride (3 in 10,000).Operating conditions—




Mobile phase: Dissolve 1.0 g of sodium lauryl sulfate in500 mL of diluted phosphoric acid (1 in 1000), and adjustthe pH to 3.0 with sodium hydroxide TS. To 240 mL of thissolution add 70 mL of tetrahydrofuran, and mix.

Flow rate: Adjust the ‰ow rate so that the retention timeof codeine is about 10 minutes.System suitability—

System performance: When the procedure is run with20 mL of the standard solution under the above operatingconditions, codeine and the internal standard are eluted inthis order with the resolution between these peaks being notless than 4.

System repeatability: When the test is repeated 5 timeswith 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratiosof the peak area of codeine to that of the internal standard isnot more than 1.0z.

10z Codeine Phosphate Powderリン酸コデイン散 10


Assay Weigh accurately about 2.5 g of 10z Codeine Phos-phate Powder, dissolve in water to make exactly 100 mL,then pipet 2 mL of this solution, add exactly 10 mL of theinternal standard solution and water to make 20 mL, anduse this solution as the sample solution. Separately, weighaccurately about 50 mg of codeine phosphate for assay,separately determined the water in the same manner asCodeine Phosphate, dissolve in water to make exactly100 mL, then pipet 10 mL of this solution, add exactly10 mL of the internal standard solution, and use this solu-tion as the standard solution. Perform the test with 20 mLeach of the sample solution and the standard solution asdirected under the Liquid Chromatography according to the


following conditions, and calculate the ratios, QT and QS, ofthe peak area of codeine to that of the internal standard:


＝WS×QT

QS× 5× 1.0227










Codeine Phosphate Tabletsリン酸コデイン錠


Assay Weigh accurately and powder not less than 20Codeine Phosphate Tablets. Weigh accurately a portion ofthe powder, equivalent to about 0.1 g of codeine phosphate(C18H21NO3.H3PO4.1/2H2O), add 30 mL of water, shake,add 20 mL of diluted dilute sulfuric acid (1 in 20), treat themixture with ultrasonic waves for 10 minutes, and add waterto make exactly 100 mL. Filter the solution, then pipet 5 mLof the ˆltrate, add exactly 10 mL of the internal standardsolution and water to make 20 mL, and use this solution asthe sample solution. Separately, weigh accurately about 50mg of codeine phosphate for assay, separately determined itswater content in the same manner as Codeine Phosphate,dissolve in water to make exactly 100 mL, then pipet 10 mL

of this solution, add exactly 10 mL of the internal standardsolution, and use this solution as the standard solution. Per-form the test with 20 mL each of the sample solution and thestandard solution as directed under the Liquid Chro-matography according to the following conditions, and cal-culate the ratios, QT and QS, of the peak area of codeine tothat of the internal standard.


＝WS×QT

QS× 2× 1.0227











Add the following:

Colistin Sulfate硫酸コリスチン

Colistin A Sulfate C53H100N16O13.212

H2SO4: 1414.66

Colistin B Sulfate C52H98N16O13.212

H2SO4: 1400.63

[1264-72-8]

Colistin Sulfate, when dried, contains not less than16,000 units per mg. The potency of Colistin Sulfate isexpressed as unit calculated from the amount ofcolistin A (C53H100N16O13: 1169.46). One unit ofColistin Sulfate is equivalent to 0.04 mg of colistin A(C53H100N16O13).

Description Colistin Sulfate occurs as a white to light yel-lowish white powder.


It is hygroscopic.

Identiˆcation (1) Dissolve 20 mg of Colistin Sulfate in2 mL of water, add 0.5 mL of sodium hydroxide TS, thenadd 5 drops of copper (II) sulfate TS while shaking: a purplecolor develops.

(2) Dissolve 50 mg of Colistin Sulfate in 10 mL of dilut-ed hydrochloric acid (1 in 2). Transfer 1 mL of this solutionin a tube for hydrolysis, seal, and heat at 1359C for 5 hours.After cooling, open the tube, and evaporate the content todryness until the odor of hydrochloric acid is no moreperceptible. Dissolve the residue in 0.5 mL of water, and usethis solution as the sample solution. Separately, dissolve20 mg each of L-leucine, L-threonine, phenylalanine and L-serine in 10 mL of water, and use these solutions as the stan-dard solution (1), the standard solution (2), the standardsolution (3) and the standard solution (4). Perform the testwith these solutions as directed under the Thin-layer Chro-matography. Spot 1 mL each of the sample solution and thestandard solutions on a plate of silica gel for thin-layer chro-matography. Develop the plate with a mixture of 1-butanol,acetic acid (100), water, pyridine and ethanol (99.5)(60:15:10:6:5) to a distance of about 10 cm, and dry the plateat 1059C for 10 minutes. Spray evenly ninhydrin TS on theplate, and heat at 1109C for 5 minutes: three principal spotsare obtained from the sample solution, the Rf values of twospots of them are the same with those of the correspondingspots obtained from the standard solution (1) and the stan-

dard solution (2), and the Rf value of the rest principal spotis about 0.1. No spot is observed at the position corre-sponding to the spots obtained from the standard solution(3) and the standard solution (4).

(3) A solution of Colistin Sulfate (1 in 20) responds tothe Qualitative Test (1) for sulfate.

Optical rotation [a]20D : －63 –－739(1.25 g, after drying,


pH The pH of a solution obtained by dissolving 0.10 g ofColistin Sulfate in 10 mL of water is between 4.0 and 6.0.

Purity (1) Sulfuric acid—Weigh accurately about 0.25 gof previously dried Colistin Sulfate, dissolve in a suitableamount of water, adjust the pH to 11 with ammonia solu-tion (28), and add water to make 100 mL. To this solutionadd exactly 10 mL of 0.1 mol/L barium chloride VS and50 mL of ethanol (99.5), and titrate with 0.1 mol/L disodi-um dihydrogen ethylenediamine tetraacetate VS until theblue-purple color of the solution disappears (indicator:0.5 mg of phthalein purple): the amount of sulfuric acid(SO4) is 16.0 to 18.0z.

Each mL of 0.1 mol/L barium chloride VS＝ 9.606 mg of SO4

(2) Related substances－Dissolve 50 mg of Colistin Sul-fate in 10 mL of water, and use this solution as the samplesolution. Pipet 1 mL of the sample solution, add water tomake exactly 50 mL, and use this solution as the standardsolution. Perform the test with these solutions as directedunder the Thin-layer Chromatography. Spot 1 mL each ofthe sample solution and the standard solution on a plate ofsilica gel for thin-layer chromatography. Develop the platewith a mixture of pyridine, 1-butanol, water and acetic acid(100) (6:5:4:1) to a distance of about 10 cm, and dry theplate at 1009C for 30 minutes. Spray evenly ninhydrin-butanol TS on the plate, and heat at 1009C for about 20minutes: the spot other than the principal spot from the sam-ple solution is not more intense than the spot from the stan-dard solution.




(1) Test organism—Escherichia coli NIHJ(2) Culture medium—Dissolve 10.0 g of peptone, 30.0 g

of sodium chloride, 3.0 g of meat extract and 15.0 g of agarin 1000 mL of water, adjust the pH with sodium hydroxideTS so that the solution will be 6.5 to 6.6 after sterilization,and use as the agar media for seed layer and for base layer.

(3) Standard solutions—Weigh accurately an amountof Colistin Sulfate Reference Standard, previously dried,equivalent to about 1,000,000 units, dissolve in phosphatebuŠer solution, pH 6.0 to make exactly 10 mL, and use thissolution as the standard stock solution. Keep the standard


stock solution at not exceeding 109C, and use within 7 days.Take exactly a suitable amount of the standard stock solu-tion before use, add phosphate buŠer solution, pH 6.0 tomake solutions so that each mL contains 10,000 units and2500 units, and use these solutions as the high concentrationstandard solution and the low concentration standard solu-tion, respectively.

(4) Sample solutions—Weigh accurately an amount ofColistin Sulfate, previously dried, equivalent to about1,000,000 units, and dissolve in phosphate buŠer solution,pH 6.0 to make exactly 10 mL. Take exactly a suitableamount of this solution, add phosphate buŠer solution, pH6.0 to make solutions so that each mL contains 10,000 unitsand 2500 units, and use these solutions as the high concen-tration sample solution and the low concentration samplesolution, respectively.


Add the following:

Daunorubicin Hydrochloride塩酸ダウノルビシン

C27H29NO10.HCl: 563.98(2S,4S )-2-Acetyl-4-(3-amino-2,3,6-trideoxy-a-L-lyxo-hexopyranosyloxy)-1,2,3,4-tetrahydro-2,5,12-trihydroxy-7-methoxynaphthacene-6,11-dionemonohydrochloride [23541-50-6]

Daunorubicin Hydrochloride contains not less than940 mg (potency) per mg, calculated on the dried basis.The potency of Daunorubicin Hydrochloride is ex-pressed as mass (potency) of daunorubicin hydrochlo-ride (C27H29NO10.HCl).

Description Daunorubicin Hydrochloride occurs as a redpowder.

It is soluble in water and in methanol, and slightly solublein ethanol (99.5).

Identiˆcation (1) Determine the absorption spectrum ofa solution of Daunorubicin Hydrochloride in methanol (1 in100,000) as directed under the Ultraviolet-visible Spec-trophotometry, and compare the spectrum with the Refer-ence Spectrum or the spectrum of a solution of Daunorubi-cin Hydrochloride Reference Standard prepared in the samemanner as the sample solution: both spectra exhibit similarintensities of absorption at the same wavelengths.

(2) A solution of Daunorubicin Hydrochloride (1 in 50)responds to the Qualitative Test (2) for chloride.

Optical rotation [a]20D : ＋250 –＋2759(15 mg calculated on

the dried basis, methanol, 10 mL, 100 mm).

pH Dissolve 0.15 g of Daunorubicin Hydrochloride in30 mL of water: the pH of the solution is between 4.5 and6.0.

Purity (1) Clarity and color of solution－Dissolve 20 mgof Daunorubicin Hydrochloride in 10 mL of water: the solu-tion is clear and red.

(2) Heavy metals—Proceed with 1.0 g of DaunorubicinHydrochloride according to Method 2, and perform the test.Prepare the control solution with 2.0 mL of Standard LeadSolution (not more than 20 ppm).

(3) Related substances—Dissolve 10 mg of Daunorubi-cin Hydrochloride in 5 mL of methanol, and use this solu-tion as the sample solution. Pipet 3 mL of the sample solu-tion, add methanol to make exactly 100 mL, and use thissolution as the standard solution. Perform the test with thesesolutions as directed under the Thin-layer Chromatography.Spot 10 mL each of the sample solution and the standardsolution on a plate of silica gel for thin-layer chromato-graphy. Develop the plate with a mixture of chloroform,methanol, water and acetic acid (100) (15:5:1:1) to a distanceof about 10 cm, and air-dry the plate. Examine the spotswith the naked eye: the spot other than the principal spotobtained from the sample solution is not more intense thanthe spot from the standard solution.


Assay Weigh accurately an amount of DaunorubicinHydrochloride and Daunorubicin Hydrochloride ReferenceStandard, equivalent to about 20 mg (potency), dissolveeach in a suitable amount of the mobile phase, add exactly4 mL of the internal standard solution and the mobile phaseto make 20 mL, and use these solutions as the sample solu-tion and the standard solution, respectively. Perform the testwith 5 mL each of the sample solution and the standard solu-tion as directed under the Liquid Chromatography accord-ing to the following conditions, and determine the ratios, QT

and QS, of the peak area of daunorubicin to that of the inter-nal standard.

Amount [mg (potency)] of C27H29NO10.HCl

＝WS×QT

QS× 1000

WS: Amount [mg (potency)] of Daunorubicin Hydrochlo-ride Reference Standard

Internal standard solution—A solution of 2-naphthalenesul-fonic acid in the mobile phase (1 in 100).Operating conditions—






Mobile phase: Adjust the pH of a mixture of water andacetonitrile (31:19) to 2.2 with phosphoric acid.

Flow rate: Adjust the ‰ow rate so that the retention timeof daunorubicin is about 9 minutes.System suitability—

System performance: When the procedure is run with 5 mLof the standard solution under the above operating condi-tions, the internal standard and daunorubicin are eluted inthis order with the resolution between these peaks being notless than 2.0.

System repeatability: When the test is repeated 6 timeswith 5 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratiosof the peak area of daunorubicin to that of the internal stan-dard is not more than 2.0z.Containers and storage Containers—Tight containers.

Add the following:

DemethylchlortetracyclineHydrochloride塩酸デメチルクロルテトラサイクリン

C21H21ClN2O8.HCl: 501.31(4S,4aS,5aS,6S,12aS )-7-Chloro-4-dimethylamino-1,4,4a,5,5a,6,11,12a-octahydro-3,6,10,12,12a-pentahydroxy-1,11-dioxonaphthacene-2-carboxamidemonohydrochloride [64-73-3]

Demethylchlortetracycline Hydrochloride containsnot less than 900 mg (potency) per mg, calculated onthe dried basis. The potency of Demethylchlortetra-cycline Hydrochloride is expressed as mass (potency)of demethylchlortetracycline hydrochloride(C21H21ClN2O8.HCl).

Description Demethylchlortetracycline Hydrochloride oc-curs as a yellow crystalline powder.

It is soluble in water, and slightly soluble in ethanol (99.5).

Identiˆcation (1) Dissolve 40 mg of Demethylchlor-tetracycline Hydrochloride in 250 mL of water. To 10 mL ofthis solution add 85 mL of water and 5 mL of a solution ofsodium hydroxide (1 in 5). Determine the absorption spec-trum of this solution as directed under the Ultraviolet-visibleSpectrophotometry, and compare the spectrum with the

Reference Spectrum or the spectrum of a solution of De-methylchlortetracycline Hydrochloride Reference Standardprepared in the same manner as the sample solution: bothspectra exhibit similar intensities of absorption at the samewavelengths.

(2) Determine the infrared absorption spectrum of De-methylchlortetracycline Hydrochloride as directed in thepotassium chloride disk method under the Infrared Spec-trophotometry, and compare the spectrum with the Refer-ence Spectrum or the spectrum of DemethylchlortetracyclineHydrochloride Reference Standard: both spectra exhibitsimilar intensities of absorption at the same wave numbers.

(3) A solution of Demethylchlortetracycline Hydrochlo-ride (1 in 100) responds to the Qualitative Test (2) for chlo-ride.

Optical rotation [a]20D : －248 –－2639(0.25 g calculated on

the dried basis, 0.1 mol/L hydrochloric acid TS, 25 mL,100 mm).

pH Dissolve 1.0 g of Demethylchlortetracycline Hydro-chloride in 100 mL of water: the pH of the solution is be-tween 2.0 and 3.0.

Purity (1) Heavy metals—Proceed with 1.0 g of De-methylchlortetracycline Hydrochloride according to Method2, and perform the test. Prepare the control solution with2.0 mL of Standard Lead Solution (not more than 20 ppm).

(2) Arsenic—Prepare the test solution with 1.0 g of De-methylchlortetracycline Hydrochloride according to Method4, and perform the test (not more than 2 ppm).

(3) Related substances—Dissolve 25 mg of Demethyl-chlortetracycline Hydrochloride in 50 mL of 0.01 mol/Lhydrochloric acid TS, and use this solution as the sample so-lution. Pipet 5 mL of the sample solution, add 0.01 mol/Lhydrochloric acid TS to make exactly 100 mL, and use thissolution as the standard solution. Perform the test with20 mL each of the sample solution and the standard solutionas directed under the Liquid Chromatography accordingto the following conditions. Determine each peak areaobtained from the chromatograms of these solutions by theautomatic integration method: the peak area other thandemethylchlortetracycline obtained from the sample solu-tion is not more than 6/5 times that of demethylchlortetracy-cline from the standard solution, and the sum of the areas ofthe peaks other than demethylchlortetracycline from thesample solution is not more than 2 times the peak area ofdemethylchlortetracycline from the standard solution.Operating conditions—


Time span of measurement: About 2 times as long as theretention time of demethylchlortetracycline after the solventpeak.System suitability—

Test for required detectability: Measure exactly 10 mL ofthe standard solution, add 0.01 mol/L hydrochloric acid TSto make exactly 50 mL, and use this solution as the solution


for system suitability test. Pipet 5 mL of the solution for sys-tem suitability test, and add 0.01 mol/L hydrochloric acidTS to make exactly 50 mL. Conˆrm that the peak area of de-methylchlortetracycline obtained from 20 mL of this solutionis equivalent to 7 to 13z of that from 20 mL of the solutionfor system suitability test.


System repeatability: When the test is repeated 6 timeswith 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peakarea of demethylchlortetracycline is not more than 1.0z.



Assay Weigh accurately an amount of Demethylchlor-tetracycline Hydrochloride and DemethylchlortetracyclineHydrochloride Reference Standard, equivalent to about25 mg (potency), dissolve each in 0.01 mol/L hydrochloricacid TS to make exactly 50 mL, and use these solutions asthe sample solution and the standard solution, respectively.Perform the test with exactly 20 mL each of the sample solu-tion and the standard solution as directed under the LiquidChromatography according to the following conditions, anddetermine the peak areas, AT and AS, of demethylchlor-tetracycline.

Amount [mg (potency)] of C21H21ClN2O8.HCl

＝WS×AT

AS× 1000

WS: Amount [mg (potency)] of DemethylchlortetracyclineHydrochloride Reference Standard



ameter and 25 cm in length, packed with styrene-divinylben-zene copolymer for liquid chromatography (10 mm in parti-cle diameter).


Mobile phase: Dissolve 3.5 g of dipotassium hydrogenphosphate, 1.5 g of tetrabutylammonium hydrogensulfateand 0.4 g of disodium dihydrogen ethylenediamine tetraa-cetate dihydrate in 300 mL of water, and adjust the pH to8.5 with sodium hydroxide TS. To this solution add 75.0 gof t-butanol and water to make 1000 mL.

Flow rate: Adjust the ‰ow rate so that the retention timeof demethylchlortetracycline is about 8 minutes.System suitability—

System performance: Heat 10 mL of the standard solutionon a water bath for 60 minutes. When the procedure is runwith 20 mL of this solution so obtained under the aboveoperating conditions, 4-epidemethylchlortetracycline anddemethylchlortetracycline are eluted in this order with theresolution between these peaks being not less than 3. The rel-

ative retention time of 4-epidemethylchlortetracycline withrespect to demethylchlortetracycline is about 0.7.

System repeatability: When the test is repeated 6 timeswith 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peakarea of demethylchlortetracycline is not more than 1.0z.


Dibekacin Sulfate硫酸ジベカシン


Dibekacin Sulfate contains not less than 640 mg(potency) per mg, calculated on the dried basis. Thepotency of Dibekacin Sulfate is expressed as mass(potency) of dibekacin (C18H37N5O8: 451.52).

Description Dibekacin Sulfate occurs as a white to yellow-ish white powder.

It is very soluble in water, and practically insoluble inethanol (99.5).

Identiˆcation (1) Dissolve 20 mg each of Dibekacin Sul-fate and Dibekacin Sulfate Reference Standard in 1 mL ofwater, and use these solutions as the sample solution and thestandard solution. Perform the test with these solutions asdirected under the Thin-layer Chromatography. Spot 5 mLeach of the sample solution and the standard solution on aplate of silica gel for thin-layer chromatography. Developthe plate with a mixture of ammonia solution (28) andmethanol (1:1) to a distance of about 10 cm, and air-dry theplate. Spray evenly 0.2z ninhydrin-water saturated 1-butanol TS, and heat at 1009C for 10 minutes: the principalspots obtained from the sample solution and the standardsolution show a purple-brown color and the same R f value.

(2) To 5 mL of a solution of Dibekacin Sulfate (1 in 50)add 1 drop of barium chloride TS: a white precipitate is pro-duced.



pH The pH of a solution obtained by dissolving 1.0 g ofDibekacin Sulfate in 20 mL of water is between 6.0 and 8.0.

Purity (1) Clarity and color of solution—Dissolve 1.0 gof Dibekacin Sulfate in 10 mL of water: the solution is clearand colorless to pale yellow.

(2) Heavy metals—Proceed with 1.0 g of Dibekacin Sul-fate according to Method 1, and perform the test. Preparethe control solution with 2.0 mL of Standard Lead Solution(not more than 20 ppm).






(3) Standard solutions－Weigh accurately an amount ofDibekacin Sulfate Reference Standard, previously dried,equivalent to about 20 mg (potency), dissolve in dilutedphosphate buŠer solution, pH 6.0 (1 in 2) to make exactly50 mL, and use this solution as the standard stock solution.Keep the standard stock solution at 5 to 159C and use within30 days. Take exactly a suitable amount of the standardstock solution before use, add 0.1 mol/L phosphate buŠersolution, pH 8.0 to make solutions so that each mL contains20 mg (potency) and 5 mg (potency), and use these solutionsas the high concentration standard solution and the low con-centration standard solution, respectively.

(4) Sample solutions—Weigh accurately an amount ofDibekacin Sulfate, equivalent to about 20 mg (potency), anddissolve in water to make exactly 50 mL. Take exactly a suit-able amount of this solution, add 0.1 mol/L phosphatebuŠer solution, pH 8.0 to make solutions so that each mLcontains 20 mg (potency) and 5 mg (potency), and use thesesolutions as the high concentration sample solution and thelow concentration sample solution, respectively.


Diclofenamideジクロフェナミド


Purity(4) Related substances—Dissolve 0.10 g of Diclofena-

mide in 50 mL of the mobile phase, and use this solution asthe sample solution. Pipet 2 mL of the sample solution, addthe mobile phase to make exactly 100 mL, and use this solu-tion as the standard solution. Perform the test with exactly10 mL each of the sample solution and the standard solutionas directed under the Liquid Chromatography according tothe following conditions, and determine each peak area bythe automatic integration method: the total area of the peaksother than the peak of diclofenamide from the sample solu-tion is not larger than the peak area of diclofenamide fromthe standard solution.Operating conditions—


Time span of measurement: About 5 times as long as theretention time of diclofenamide.System suitability—

Test for required detection: To exactly 5 mL of the stand-ard solution add the mobile phase to make exactly 100 mL.

Conˆrm that the peak area of diclofenamide obtained from10 mL of this solution is equivalent to 3.5 to 6.5z of that ofdiclofenamide obtained from 10 mL of the standard solu-tion.


System repeatability: When the test is repeated 6 timeswith 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peakarea of diclofenamide is not more than 1.0z.


Assay Weigh accurately about 50 mg each of Diclofena-mide and Diclofenamide Reference Standard, previouslydried, and dissolve each in 30 mL of the mobile phase. Toeach add exactly 10 mL of the internal standard solution andthe mobile phase to make 50 mL, and use these solutions asthe sample solution and the standard solution. Perform thetest with 10 mL each of the sample solution and the standardsolution as directed under the Liquid Chromatography ac-cording to the following conditions, and calculate the ratios,QT and QS, of the peak area of diclofenamide to that of theinternal standard, respectively.

Amount (mg) of C6H6Cl2N2O4S2＝WS×QT

QS

WS: Amount (mg) of Diclofenamide Reference Standard

Internal standard solution—A solution of butyl parahydroxybenzoate in the mobile phase (3 in 5000).Operating conditions—




Mobile phase: A mixture of sodium phosphate TS andacetonitrile (1:1).

Flow rate: Adjust the ‰ow rate so that the retention timeof diclofenamide is about 7 minutes.System suitability—

System performance: When the procedure is run with10 mL of the standard solution under the above operatingconditions, diclofenamide and the internal standard are elut-ed in this order with the resolution between these peaksbeing not less than 9.

System repeatability: When the test is repeated 6 timeswith 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratiosof the peak area of diclofenamide to that of the internalstandard is not more than 1.0z.


Diethylcarbamazine Citrate Tabletsクエン酸ジエチルカルバマジン錠


Assay Weigh accurately and powder not less than 20Diethylcarbamazine Citrate Tablets. Weigh accurately aportion of the powder, equivalent to about 50 mg of diethyl-carbamazine citrate (C10H21N3O.C6H8O7), add 10 mL ofwater, shake well, add 5 mL of sodium hydroxide TS, thenadd exactly 20 mL of the internal standard solution, andshake vigorously for 10 minutes. Centrifuge, discard theaqueous layer, and use the chloroform layer as the samplesolution. Separately, weigh accurately about 50 mg ofDiethylcarbamazine Citrate Reference Standard, previouslydried at 1059C for 4 hours, dissolve in 10 mL of water, add5 mL of sodium hydroxide TS, proceed in the same manneras the preparation of the sample solution, and use the chlo-roform layer as the standard solution. Perform the test with2 mL of the sample solution and the standard solution asdirected under the Gas Chromatography according to thefollowing conditions, and calculate the ratios, QT and QS, ofthe peak area of diethycarbamazine to that of the internalstandard, respectively.

Amount (mg) of diethylcarbamazine citrate(C10H21N3O.C6H8O7)

＝WS×QT

QS

WS: Amount (mg) of Diethylcarbamazine Citrate Refer-ence Standard

Internal standard solution—A solution of n-octadecane inchloroform (1 in 1250).Operating conditions—

Detector: A hydrogen ‰ame-ionization detector.Column: A glass tube 3 mm in inside diameter and 1 m in

length, packed with silanized siliceous earth for gas chro-matography (180 to 250 mm in particle diameter) coated with35z methylphenyldimethyl silicone polymer for gas chro-matography in the ratio of 3z.



of diethylcarbamazine is about 4 minutes.System suitability—

System performance: When the procedure is run with 2 mLof the standard solution under the above operating condi-tions, diethylcarbamazine and the internal standard are elut-ed in this order with the resolution between these peaksbeing not less than 5.

System repeatability: When the test is repeated 6 timeswith 2 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratiosof the peak area of diethylcarbamazine to that of the inter-nal standard is not more than 1.5z.

1z Dihydrocodeine PhosphatePowderリン酸ジヒドロコデイン散 1


Assay Weigh accurately about 5 g of 1z DihydrocodeinePhosphate Powder, dissolve in water to make exactly100 mL, then pipet 10 mL of this solution, add exactly10 mL of the internal standard solution, and use this solu-tion as the sample solution. Separately, weigh accuratelyabout 50 mg of dihydrocodeine phosphate for assay,separately determined its loss on drying (1059C, 4 hours),dissolve in water to make exactly 100 mL, then pipet 10 mLof this solution, add exactly 10 mL of the internal standardsolution, and use this solution as the standard solution. Per-form the test with 20 mL each of the sample solution and thestandard solution as directed under the Liquid Chro-matography according to the following conditions, andcalculate the ratios, QT and QS, of the peak area of dihydro-codeine to that of the internal standard.

Amount (mg) of dihydrocodeine phosphate(C18H23NO3.H3PO4)

＝WS×QT

QS

WS: Amount (mg) of dihydrocodeine phosphate for as-say, calculated on the dried basis

Internal standard solution—A solution of ethylefurinhydrochloride (3 in 10,000).Operating conditions—




Mobile phase: Dissolve 1.0 g of sodium lauryl sulfate in500 mL of diluted phosphoric acid (1 in 1000), and adjustthe pH to 3.0 with sodium hydroxide TS. To 240 mL of thissolution add 70 mL of tetrahydrofuran.

Flow rate: Adjust the ‰ow rate so that the retention timeof dihydrocodeine is about 9 minutes.System suitability—

System performance: When the procedure is run with20 mL of the standard solution under the above operatingconditions, dihydrocodeine and the internal standard areeluted in this order with the resolution between these peaksbeing not less than 4.

System repeatability: When the test is repeated 5 timeswith 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratiosof the peak area of dihydrocodeine to that of the internalstandard is not more than 1.0z.


10z Dihydrocodeine PhosphatePowderリン酸ジヒドロコデイン散 10


Assay Weigh accurately about 2.5 g of 10z Dihydro-codeine Phosphate Powder, dissolve in water to make ex-actly 100 mL, then pipet 2 mL of this solution, add exactly10 mL of the internal standard solution and water to make20 mL, and use this solution as the sample solution.Separately, weigh accurately about 50 mg of dihydrocodeinephosphate for assay, separately determined its loss on drying(1059C, 4 hours), dissolve in water to make exactly 100 mL,then pipet 10 mL of this solution, add exactly 10 mL of theinternal standard solution, and use this solution as the stan-dard solution. Perform the test with 20 mL each of the sam-ple solution and the standard solution as directed under theLiquid Chromatography according to the following condi-tions, and calculate the ratios, QT and QS, of the peak areaof dihydrocodeine to that of the internal standard.

Amount (mg) of dihydrocodeine phosphate(C18H23NO3.H3PO4)

＝WS×QT

QS× 5

WS: Amount (mg) of dihydrocodeine phosphate for as-say, calculated on the dried basis

Internal standard solution—A solution of ethylefrinehydrochloride (3 in 10,000).Operating conditions—




Mobile phase: Dissolve 1.0 g of sodium lauryl sulfate in500 mL of diluted phosphoric acid (1 in 1000), and adjustthe pH to 3.0 with sodium hydroxide TS. To 240 mL of thissolution add 70 mL of tetrahydrofuran.

Flow rate: Adjust the ‰ow rate so that the retention timeof dihydrocodeine is about 9 minutes.System suitability—

System performance: When the procedure is run with20 mL of the standard solution under the above operatingconditions, dihydrocodeine and the internal standard areeluted in this order with the resolution between these peaksbeing not less than 4.

System repeatability: When the test is repeated 5 timeswith 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratiosof the peak area of dihydrocodeine to that of the internalstandard is not more than 1.0z.

Dihydroergotoxine Mesilateメシル酸ジヒドロエルゴトキシン


Assay (1) Dihydroergotoxine mesilate—Weigh accurate-ly about 30 mg each of Dihydroergotoxine Mesilate and Di-hydroergotoxine Mesilate Reference Standard, and dissolvethem separately in a suitable amount of a mixture of waterand acetonitrile (3:1). To these solutions add exactly 10 mLof the internal standard solution and an amount of a mixtureof water and acetonitrile (3:1) to make 50 mL, and use thesesolutions as the sample solution and the standard solution.Perform the test with 20 mL of the sample solution and thestandard solution as directed under the Liquid Chro-matography according to the following conditions, and cal-culate the ratios of the peak areas of dihydroergocornine, di-hydro-a-ergocryptine, dihydroergocristine and dihydro-b-ergocryptine to the peak area of the internal standard ofthese solutions.

Amount (mg) of dihydroergotoxine mesilate

＝WS×MTA＋MTB＋MTC＋MTD

MSA＋MSB＋MSC＋MSD

WS: Amount (mg) of Dihydroergotoxine Mesilate Refer-ence Standard, calculated on the anhydrous basis

MTA: Ratio of the peak area of dihydroergocornine tothat of the internal standard of the sample solution× 659.80

MTB: Ratio of the peak area of dihydro-a-ergocryptine tothat of the internal standard of the sample× 673.83

MTC: Ratio of the peak area of dihydroergocristine to thatof the internal standard of the sample solution ×707.85

MTD: Ratio of the peak area of dihydro-b-ergocryptine tothat of the internal standard of the sample solution× 673.83

MSA: Ratio of the peak area of dihydroergocornine to thatof the internal standard of the standard solution ×659.80

MSB: Ratio of the peak area of dihydro-a-ergocryptine tothat of the internal standard of the standard solution× 673.83

MSC: Ratio of the peak area of dihydroergocristine to thatof the internal standard of the standard solution ×707.85

MSD: Ratio of the peak area of dihydro-b-ergocryptine tothat of the internal standard of the standard solu-tion × 673.83

Internal standard solution—Dissolve 0.04 g of chloram-phenicol in a mixture of water and acetonitrile (3:1) to make250 mL.Operating conditions—






Mobile phase: A mixture of water, acetonitrile andtriethylamine (30:10:1).

Flow rate: Adjust the ‰ow rate so that the retention timeof chloramphenicol is about 5 minutes.System suitability—

System performance: When the procedure is run with20 mL of the standard solution under the above operatingconditions, the internal standard, dihydroergocornine, di-hydro-a-ergocryptine, dihydroergocristine and dihydro-b-ergocryptine are eluted in this order with the resolutionbetween the peaks of dihydro-a-ergocryptine and dihydroer-gocristine being not less than 1.5.

System repeatability: When the test is repeated 6 timeswith 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratiosof the peak area of dihydroergocornine, dihydro-a-ergocryptine, dihydroergocristine and dihydro-b-ergocryp-tine to that of the internal standard is not more than 0.5z.

Diltiazem Hydrochloride塩酸ジルチアゼム


Purity(5) Related substances—Dissolve 50 mg of Diltiazem

Hydrochloride in 50 mL of diluted ethanol (99.5) (4 in 5),and use this solution as the sample solution. Measure exactly1 mL of the sample solution, add diluted ethanol (99.5) (4 in5) to make exactly 200 mL, and use this solution as the stan-dard solution. Perform the test with exactly 20 mL each ofthe sample solution and the standard solution as directed un-der the Liquid Chromatography according to the followingconditions. Determine each peak area of both solutions byautomatic integration method: the total peak area of peaksother than the peak of diltiazem obtained from the samplesolution is not more than 3/5 times the peak area of diltia-zem obtained from the standard solution.Operating conditions—


Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silicagel for liquid chromatography (5 mm in particle diameter).


Mobile phase: Dissolve 8 g of sodium acetate trihydrateand 1.5 g of d-camphorsulfonic acid in 500 mL of water,and ˆlter using a membrane ˆlter (0.4 mm in pore size). Add250 mL each of acetonitrile and methanol to the ˆltrate, andadjust the solution to a pH of 6.6 by adding sodium acetatetrihydrate.

Flow rate: Adjust the ‰ow rate so that the retention timeof diltiazem is about 9 minutes.

Time span of measurement: About twice as long as theretention time of diltiazem after the solvent peak.System suitability—

Test for required detection: To exactly 2 mL of the stan-dard solution add diluted ethanol (99.5) (4 in 5) to makeexactly 10 mL. Conˆrm that the peak area of diltiazemobtained from 20 mL of this solution is equivalent to 15 to25z of that of diltiazem obtained from 20 mL of the stan-dard solution.

System performance: Dissolve 0.03 g of DiltiazemHydrochloride, 0.02 g of d-3-hydroxy-cis-2,3-dihydro-5-[2-(dimethylamino)ethyl]-2-(4-methoxyphenyl)-1, 5-benzothia-zepin-4-(5H)-one hydrochloride and 0.02 g of phenylbenzo-ate in 160 mL of ethanol (99.5), and add water to make 200mL. Perform the test with 20 mL of this solution as directedunder the Liquid Chromatography under the above operat-ing conditions: d-3-hydroxy-cis-2,3-dihydro-5-[2-(dimethyl-amino)ethyl] - 2 - (4 - methoxyphenyl) - 1, 5 - benzothiazepin-4(5H )-one, diltiazem and phenyl benzoate are eluted in thisorder with the resolutions between the peaks of d-3-hydroxy-cis - 2, 3 - dihydro - 5 - [2 - (dimethylamino)ethyl] - 2 - (4 -methoxyphenyl)-1,5-benzothiazepin-4(5H )-one and diltia-zem and between the peaks of diltiazem and phenyl benzoatebeing not less than 2.5, respectively.

System repeatability: When the test is repeated 6 timeswith 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peakarea of diltiazem is not more than 2.0z.

Dipyridamoleジピリダモール


Purity(4) Related substances—Dissolve 50 mg of Dipyrida-

mole in 50 mL of the mobile phase, and use this solution asthe sample solution. Pipet 0.5 mL of the sample solution,add the mobile phase to make exactly 100 mL, and use thissolution as the standard solution. Perform the test with ex-actly 20 mL each of the sample solution and the standard so-lution as directed under the Liquid Chromatography accord-ing to the following conditions, and determine each peakarea by the automatic integration method: the total area ofthe peaks other than the peak of dipyridamole from the sam-ple solution is not larger than the peak area of dipyridamolefrom the standard solution.Operating conditions—



Column temperature: A constant temperature of about


409C.Mobile phase: Dissolve 0.2 g of potassium dihydrogen

phosphate in 200 mL of water, and add 800 mL ofmethanol.

Flow rate: Adjust the ‰ow rate so that the retention timeof dipyridamole is about 4 minutes.

Time span of measurement: About 5 times as long as theretention time of dipyridamole.System suitability—

Test for required detection: To exactly 5 mL of the stan-dard solution add the mobile phase to make exactly 25 mL.Conˆrm that the peak area of dipyridamole obtained from20 mL of this solution is equivalent to 15 to 25z of that ofdipyridamole obtained from 20 mL of the standard solution.

System performance: Dissolve 7 mg of Dipyridamole and3 mg of terphenyl in 50 mL of methanol. When the proce-dure is run with 20 mL of this solution under the above oper-ating conditions, dipyridamole and terphenyl are eluted inthis order with the resolution between these peaks being notless than 5.

System repeatability: When the test is repeated 6 timeswith 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peakarea of dipyridamole is not more than 1.0z.

Dobutamine Hydrochloride塩酸ドブタミン

Change the pH to read:

pH Dissolve 1.0 g of Dobutamine Hydrochloride in100 mL of water: the pH of this solution is between 4.0 and5.5.


Assay Weigh accurately about 0.1 g each of DobutamineHydrochloride and Dobutamine Hydrochloride ReferenceStandard, each previously dried, dissolve separately in ex-actly 10 mL of the internal standard solution, add dilutedmethanol (1 in 2) to make 50 mL, and use these solutions asthe sample solution and the standard solution respectively.Perform the test with 5 mL each of the sample solution andthe standard solution as directed under the Liquid Chro-matography according to the following conditions, and cal-culate the ratios, QT and QS, of the peak area of dobutamineto that of the internal standard, respectively.

Amount (mg) of C18H23NO3.HCl＝WS×QT

QS

WS: Amount (mg) of Dobutamine Hydrochloride Refer-ence Standard

Internal standard solution—A solution of salicylamide indiluted methanol (1 in 2) (1 in 125).Operating conditions—





Mobile phase: A mixture of tartrate buŠer solution, pH3.0 and methanol (7:3).

Flow rate: Adjust the ‰ow rate so that the retention timeof dobutamine is about 7 minutes.System suitability—

System performance: When the procedure is run with 5 mLof the standard solution under the above operating condi-tions, dobutamine and internal standard are eluted in thisorder with the resolution between these peaks being not lessthan 5.

System repeatability: When the test is repeated 6 timeswith 5 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratiosof the peak area of dobutamine to that of the internal stan-dard is not more than 1.0z.

Dopamine Hydrochloride Injection塩酸ドパミン注射液


Assay To an exact volume of Dopamine HydrochlorideInjection, equivalent to about 30 mg of dopaminehydrochloride (C8H11NO2.HCl), add the mobile phase tomake exactly 50 mL. Pipet 2.5 mL of this solution, addexactly 2.5 mL of the internal standard solution and themobile phase to make 20 mL, and use this solution as thesample solution. Separately, weigh accurately about 30 mgof dopamine hydrochloride for assay, previously dried at1059C for 3 hours, dissolve in the mobile phase to makeexactly 50 mL. Pipet 2.5 mL of this solution, add exactly2.5 mL of the internal standard solution and the mobilephase to make 20 mL, and use this solution as the standardsolution. Perform the test with 10 mL each of the samplesolution and the standard solution as directed under the Liq-uid Chromatography according to the following conditions,and calculate the ratios, QT and QS, of the peak area ofdopamine to that of the internal standard.

Amount (mg) of dopamine hydrochloride (C8H11NO2.HCl)

＝WS×QT

QS

WS: Amount (mg) of dopamine hydrochloride for assay

Internal standard solution—A solution of uracil in themobile phase (3 in 10,000).Operating conditions—


Column: A stainless steel column 4.6 mm in inside di-




Mobile phase: Disodium hydrogen phosphate-citric acidbuŠer solution, pH 3.0

Flow rate: Adjust the ‰ow rate so that the retention timeof dopamine is about 10 minutes.System suitability—

System performance: When the procedure is run with10 mL of the standard solution under the above operatingconditions, the internal standard and dopamine are eluted inthis order with the resolution between these peaks being notless than 10.

System repeatability: When the test is repeated 6 timeswith 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratiosof peak area of dopamine to that of the internal standard isnot more than 1.0z.



Doxorubicin Hydrochloride塩酸ドキソルビシン


Doxorubicin Hydrochloride contains not less than980 mg (potency) per mg, calculated on the anhydrousbasis. The potency of Doxorubicin Hydrochlorideis expressed as mass (potency) of doxorubicinhydrochloride (C27H29NO11. HCl).

Description Doxorubicin Hydrochloride occurs as a red-orange crystalline powder.

It is sparingly soluble in water, slightly soluble inmethanol, very slightly soluble in ethanol (99.5), and practi-cally insoluble in acetonitrile.

Identiˆcation (1) Determine the absorption spectrum ofa solution of Doxorubicin Hydrochloride in methanol (1 in100,000) as directed under the Ultraviolet-visible Spec-trophotometry, and compare the spectrum with the Refer-ence Spectrum or the spectrum of a solution of DoxorubicinHydrochloride Reference Standard prepared in the samemanner as the sample solution: both spectra exhibit similarintensities of absorption at the same wavelengths.

(2) Determine the infrared absorption spectrum of Dox-orubicin Hydrochloride as directed in the potassium chloridedisk method under the Infrared Spectrophotometry, andcompare the spectrum with the Reference Spectrum or thespectrum of Doxorubicin Hydrochloride Reference Stan-

dard: both spectra exhibit similar intensities of absorption atthe same wave numbers.

(3) A solution of Doxorubicin Hydrochloride (1 in 200)responds to the Qualitative Test (1) for chloride.


the anhydrous basis, methanol, 20 mL, 100 mm).

pH The pH of a solution obtained by dissolving 50 mg ofDoxorubicin Hydrochloride in 10 mL of water is between4.0 and 5.5.

Purity (1) Clarity and color of solution—Dissolve 50 mgof Doxorubicin Hydrochloride in 10 mL of water: the solu-tion is clear and red.

(2) Related substances—Dissolve 25 mg of DoxorubicinHydrochloride in 100 mL of the mobile phase, and use thissolution as the sample solution. Pipet 2 mL of the samplesolution, add the mobile phase to make exactly 100 mL, anduse this solution as the standard solution. Perform the testwith exactly 20 mL each of the sample solution and the stan-dard solution as directed under the Liquid Chromatographyaccording to the following conditions, and determine eachpeak area by the automatic integration method: the area ofthe peak other than doxorubicin obtained from the samplesolution is not more than 1/4 times the peak area of dox-orubicin from the standard solution, and the total area ofthe peaks other than doxorubicin is not more than the peakarea of doxorubicin from the standard solution.Operating conditions—




Mobile phase: Dissolve 3 g of sodium lauryl sulfate in1000 mL of diluted phosphoric acid (7 in 5000), and add1000 mL of acetonitrile.

Flow rate: Adjust the ‰ow rate so that the retention timeof doxorubicin is about 8 minutes.

Time span of measurement: About 3 times as long as theretention time of doxorubicin.System suitability—

Test for required detectability: Measure 1 mL of the stan-dard solution, and add the mobile phase to make exactly20 mL. Conˆrm that the peak area of doxorubicin obtainedfrom 20 mL of this solution is equivalent to 3.5 to 6.5z ofthat from 20 mL of the standard solution.

System performance: Dissolve 5 mg of DoxorubicinHydrochloride in 20 mL of water, add 1.5 mL of phosphoricacid, and allow to stand at room temperature for 30minutes. Adjust the pH of this solution to 2.5 with 2 mol/Lsodium hydroxide TS. When the procedure is run with 20 mLof this solution under the above operating conditions, dox-orubicinone, having the relative retention time of about 0.6with respect to doxorubicin, and doxorubicin are eluted in


this order with the resolution between these peaks being notless than 5.

System repeatability: When the test is repeated 6 timeswith 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peakarea of doxorubicin is not more than 2.0z.


Assay Weigh accurately an amount of DoxorubicinHydrochloride and Doxorubicin Hydrochloride ReferenceStandard, equivalent to about 10 mg (potency), dissolveeach in water to make exactly 25 mL. Pipet 5 mL each ofthese solutions, add water to make exactly 100 mL, and usethese solutions as the sample solution and the standard solu-tion, respectively. Determine the absorbances at 495 nm, AT

and AS, of the sample solution and the standard solution asdirected under the Ultraviolet-visible Spectrophotometry.


＝WS×AT

AS× 1000

WS: Amount [mg (potency)] of Doxorubicin Hydrochlo-ride Reference Standard


Doxycycline Hydrochloride塩酸ドキシサイクリン


Doxycycline Hydrochloride contains not less than880 mg (potency) per mg, calculated on the anhydrousbasis and corrected by the amount of ethanol. Thepotency of Doxycycline Hydrochloride is expressed asmass (potency) of doxycycline (C22H24N2O8: 444.43).

Description Doxycycline Hydrochloride occurs as yellowto dark yellow, crystals or crystalline powder.

It is freely soluble in water and in methanol, and slightlysoluble in ethanol (99.5).

Identiˆcation (1) Determine the infrared absorptionspectrum of Doxycycline Hydrochloride as directed in thepotassium bromide disk method under the Infrared Spec-trophotometry, and compare the spectrum with the Refer-ence Spectrum or the spectrum of Doxycycline Hydrochlo-ride Reference Standard: both spectra exhibit similar intensi-ties of absorption at the same wave numbers.

(2) Dissolve 10 mg of Doxycycline Hydrochloride in10 mL of water, and add silver nitrate TS: a white turbidityis produced.

Absorbance E 1z1 cm (349 nm): 285 – 315 (10 mg, 0.01 mol/L

hydrochloric acid-methanol TS, 500 mL).

Optical rotation [a]20D : －105 –－1209 (0.25 g calculated

on the anhydrous basis and correcred by the amount ofethanol, 0.01 mol/L hydrochloric acid-methanol TS, 25 mL,100 mm). Determine within 5 minutes after the sample solu-tion is prepared.

Purity (1) Heavy metals—Proceed with 1.0 g of Doxycy-cline Hydrochloride according to Method 2, and performthe test. Prepare the control solution with 5.0 mL of Stan-dard Lead Solution (not more than 50 ppm).

(2) Related substance—Dissolve 20 mg of DoxycyclineHydrochloride in 0.01 mol/L hydrochloric acid TS to makeexactly 25 mL, and use this solution as the sample solution.Separately, dissolve 20 mg of 6-epidoxycycline hydrochlo-ride in 0.01 mol/L hydrochloric acid TS to make exactly25 mL, and use this solution as 6-epidoxycycline hydrochlo-ride stock solution. Separately, dissolve 20 mg of metacy-cline hydrochloride in 0.01 mol/L hydrochloric acid TS tomake exactly 25 mL, and use this solution as metacyclinehydrochloride stock solution. Pipet 2 mL each of 6-epidoxy-cycline hydrochloride stock solution and metacyclinehydrochloride stock solution, add 0.01 mol/L hydrochloricacid TS to make exatly 100 mL, and use this solution as thestandard solution. Perform the test with exactly 20 mL eachof the sample solution and the standard solution as directedunder the Liquid Chromatography according to the follow-ing conditions, and determine each peak area by the auto-matic integration method: the peak areas of metacycline and6-epidoxycycline obtained from the sample solution are notmore than the peak areas of them obtained from the stan-dard solution, respectively, and the areas of the two peaks,appeared between the solvent peak and metacycline andbehind of doxycycline, obtained from the sample solutionare not more than 1/4 of the peak area of 6-epidoxycyclinefrom the standard solution.Operating conditions—


Column: A stainless steel column 4.6 mm in inside di-ameter and 25 cm in length, packed with styrene-divinylben-zene copolymer for liquid chromatography (8 mm in particlediameter).


Mobile phase: Mix 125 mL of 0.2 mol/L potassiumdihydrogen phosphate TS, 117 mL of 0.2 mol/L sodiumhydroxide TS, and add water to make 500 mL. To 400 mLof this solution add 50 mL of a solution of tetrabutylammo-nium hydrogensulfate (1 in 100), 10 mL of a solution of dis-odium dihydrogen ethylenediamine tetraacetate dihydrate (1in 25), 60 g of t-butanol and 200 mL of water, adjust the pHto 8.0 with 2 mol/L sodium hydroxide TS, and add water tomake 1000 mL.

Flow rate: Adjust the ‰ow rate so that the retention timeof doxycycline is about 19 minutes.

Time span of measurement: About 2.4 times as long as theretention time of doxycycline after the solvent peak.



the standard solution, and add 0.01 mol/L hydrochloricacid TS to make exactly 20 mL. Conˆrm that the peak areasof 6-epidoxycycline and metacycline obtained from 20 mL ofthis solution are equivalent to 3.5 to 6.5z of them obtainedfrom 20 mL of the standard solution, respectively.

System performance: To 8 mL of the sample solution,3 mL of 6-epidoxycycline hydrochloride stock solution and2 mL of metacycline hydrochloride stock solution add0.01 mol/L hydrochloric acid TS to make 50 mL. When theprocedure is run with 20 mL of this solution under the aboveoperating conditions, metacycline, 6-epidoxycycline anddoxycycline are eluted in this order with the resolutionsbetween the peaks, metacycline and 6-epidoxycycline, and 6-epidoxycycline and doxycycline, being not less than 1.3 andnot less than 2.0, respectively, and the symmetry coe‹cientof the peak of doxycycline is not more than 1.3.

System repeatability: When the test is repeated 6 timeswith 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviations of the peakarea of metacycline and 6-epidoxycycline are not more than3.0z and not more than 2.0z, respectively.

Ethanol Weigh accurately about 0.1 g of DoxycyclineHydrochloride, dissolve in the internal standard solution tomake exactly 10 mL, and use this solution as the samplesolution. Separately, weigh accurately about 0.4 g of ethanol(99.5), and add the internal standard solution to makeexactly 100 mL. Pipet 1 mL of this solution, add the internalstandard solution to make exactly 10 mL, and use this solu-tion as the standard solution. Perform the test with 1 mLeach of the sample solution and the standard solution asdirected under the Gas Chromatography according to thefollowing conditions, and determine the ratios, QT and QS,of the peak area of ethanol to that of the internal standard:the amount of ethanol is not less than 4.3z and not morethan 6.0z.

Amount (z) of ethanol＝WS

WT×

QT

QS

WS: Amount (mg) of ethanol (99.5)WT: Amount (mg) of the sample

Internal standard solution—A solution of 1-propanol (1 in2000).Operating conditions—

Detector: An hydrogen ‰ame-ionization detector.Column: A glass column 3.2 mm in inside diameter

and 1.5 m in length, packed with porous ethylvinylbenzene-divinylbenzene copolymer for gas chromatography(0.0075 mm in average pore size, 500 – 600 m2/g in speciˆcsurface area) (150 – 180 mm in particle diameter).



of ethanol is about 5 minutes.

System suitability—System performance: When the procedure is run with 1 mL

of the standard solution under the above operating condi-tions, ethanol and the internal standard are eluted in thisorder with the resolution between these peaks being not lessthan 2.0.

System repeatability: When the test is repeated 5 timeswith 1 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratiosof the peak area of ethanol to that of the internal standard isnot more than 2.0z.

Water Not less than 1.4z and not more than 2.8z (0.6 g,volumetric titration, direct titration).


Assay Weigh accurately an amount of DoxycyclineHydrochloride and Doxycycline Hydrochloride ReferenceStandard, equivalent to about 50 mg (potency), dissolveeach in water to make exactly 50 mL, and use these solutionsas the sample solution and the standard solution. Performthe test with exactly 10 mL each of the sample solution andthe standard solution as directed under the Liquid Chro-matography according to the following conditions, anddetermine the peak areas, AT and AS, of doxycycline.

Amount [mg (potency)] of doxycycline (C22H24N2O8)

＝WS×AT

AS× 1000

WS: Amount [mg (potency)] of Doxycycline Hydrochlo-ride Reference Standard





Mobile phase: Dissolve 7.0 g of sodium dihydrogen phos-phate dihydrate in 450 mL of water, add 553 mL of a mix-ture of methanol and N,N-dimethyl-n-octylamine (550:3),and adjust the pH to 8.0 with a solution of sodiumhydroxide (43 in 200).

Flow rate: Adjust the ‰ow rate so that the retention timeof doxycycline is about 6 minutes.System suitability—

System performance: When the procedure is run with10 mL of the standard solution under the above operatingconditions, the theoretical plates and the symmetricalcoe‹cient of the peak of doxycycline are not less than 1000and not more than 2.0, respectively.

System repeatability: When the test is repeated 6 timeswith 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peakarea of doxycycline is not more than 1.0z.



Droperidolドロペリドール


Description Droperidol occurs as a white to light yellowpowder.

It is freely soluble in acetic acid (100), soluble indichloromethane, slightly soluble in ethanol (99.5), andpractically insoluble in water.

It is gradually colored by light.


Identiˆcation (1) Put 30 mg of Droperidol in a brownvolumetric ‰ask, and dissolve in 10 mL of 0.1 mol/Lhydrochloric acid TS and ethanol (95) to make 100 mL.Transfer 5 mL of the solution to a brown volumetric ‰ask,and add 10 mL of 0.1 mol/L hydrochloric acid TS andethanol (95) to make 100 mL. Determine the absorptionspectrum of the solution as directed under the Ultraviolet-visible Spectrophotometry, and compare the spectrum withthe Reference Spectrum: both spectra exhibit similar intensi-ties of absorption at the same wavelengths.

(2) Determine the infrared absorption spectrum ofDroperidol, previously dried, as directed in the potassiumbromide disk method under the Infrared Spectrophotomet-ry, and compare the spectrum with the Reference Spectrum:both spectra exhibit similar intensities of absorption at thesame wave numbers. If any diŠerence appears between thespectra, dissolve Droperidol in acetone, evaporate the ace-tone, dry the residue in a desiccator (in vacuum, silica gel,709C) for 4 hours, and perform the test with the residue.

Delete the Melting point.

Delete the following Monographs:

Drostanolone Propionateプロピオン酸ドロスタノロン

Drostanolone Propionate Injectionプロピオン酸ドロスタノロン注射液

En‰uraneエンフルラン


Purity(4) Related substances—Proceed the test with 5 mL of

En‰urane as directed under the Gas chromatography ac-cording to the following conditions. Determine each peakarea other than the peak of air which appears soon after in-jection of the sample by the automatic integration method,and calculate the amount of each peak by the area percen-tage method: the amount of the substances other than en‰u-rane is not more than 0.10z.Operating conditions—

Detector: A thermal conductivity detector.Column: A column 3 mm in inside diameter and 3 m in

length, packed with siliceous earth for gas chromatography,180 to 250 mm in particle diameter, coated with diethyleneglycol succinate ester for gas chromatography in the ratio of20z.


Carrier gas: HeliumFlow rate: Adjust the ‰ow rate so that the retention time

of en‰urane is about 3 minutes.Time span of measurement: About three times as long as

the retention time of en‰urane.System suitability—

Test for required detection: Pipet exactly 1 mL of en‰u-rane add 2-propanol to make exactly 100 mL. To exactly2 mL of this solution add 2-propanol to make exactly10 mL, and use this solution as the solution for systemsuitability test. Pipet 1 mL of the solution, and add 2-propanol to make exactly 10 mL. Conˆrm that the peak areaof en‰urane obtained from 5 mL of this solution is equiva-lent to 7 to 13z of that of en‰urane obtained from 5 mL ofthe solution for system suitability test.

System performance: Mix 5 mL of En‰urane and 5 mL of2-propanol. When the procedure is run with 5 mL of thismixture under the above operating conditions, en‰urane and2-propanol are eluted in this order with the resolution be-tween these peaks being not less than 2.0.

System repeatability: When the test is repeated 6 timeswith 5 mL of the solution for system suitability test under theabove operating conditions, the relative standard deviationof the peak area of en‰urane is not more than 2.0z.


Add the following:

Enviomycin Sulfate硫酸エンビオマイシン

Tuberactinomycin N Sulfate

C25H43N13O10.112 H2SO4: 832.81

Tuberactinomycin O Sulfate

C25H43N13O9.112

H2SO4: 816.81

Tuberactinomycin N Sulfate(3R,4R )-N-[(3S,9S,12S,15S )-9,12-Bis(hydroxymethyl)-3-[(4R )-2-iminohexahydropyrimidin-4-yl]-2,5,8,11,14-pentaoxo-6-(Z )-ureidomethylene-1,4,7,10,13-pentaazacyclohexadec-15-yl]-3,6-diamino-4-hydroxyhexanamide sesquisulfate[33103-22-9, Tuberactinomycin N]

Tuberactinomycin O Sulfate(S )-N-[(3S,9S,12S,15S )-9,12-Bis(hydroxymethyl)-3-[(4R )-2-iminohexahydropyrimidin-4-yl]-2,5,8,11,14-pentaoxo-6-(Z )-ureidomethylene-1,4,7,10,13-pentaazacyclohexadec-15-yl]-3,6-diaminohexanamide sesquisulfate[33137-73-4, Tuberactinomycin O]

Enviomycin Sulfate contains not less than 770 mg(potency) per mg, calculated on the dried basis. Thepotency of Enviomycin Sulfate is expressed as mass(potency) of tuberactinomycin N (C25H43N13O10:685.69).

Description Enviomycin Sulfate occurs as a white powder.It is very soluble in water, and practically insoluble in

ethanol (99.5).

Identiˆcation (1) To 5 mL of a solution of EnviomycinSulfate (1 in 200) add 1.5 mL of sodium hydroxide TS, andadd 1 drop of a mixture of 0.01 mol/L citric acid TS andcopper (II) sulfate TS (97:3) : a blue-purple color develops.

(2) Determine the absorption spectrum of a solution ofEnviomycin Sulfate (1 in 100,000) as directed under theUltraviolet-visible Spectrophotometry, and compare thespectrum with the Reference Spectrum: both spectra exhibitsimilar intensities of absorption at the same wavelengths.

(3) To 2 mL of a solution of Enviomycin Sulfate (1 in

20) add 1 drop of barium chloride TS: a white precipitate isproduced.



pH The pH of a solution obtained by dissolving 2.0 g ofEnviomycin Sulfate in 20 mL of water is between 5.5 and7.5.

Content ratio of the active principle Dissolve 0.1 g of En-viomycin Sulfate in water to make 100 mL, and use this solu-tion as the sample solution. Perform the test with 3 mL of thesample solution as directed under the Liquid Chro-matography according to the following conditions, and de-termine the peak areas, AT1 and AT2, of tuberactinomycin Nand tuberactinomycin O, having the relative retention time,1.4± 0.4, with respect to tuberactinomycin N, by the auto-matic integration method: AT2/(AT1＋ AT2) is between 0.090and 0.150.Operating conditions—


Column: A stainless steel column 4.6 mm in inside di-ameter and 15 cm in length, packed with silica gel for liquidchromatography (5 mm in particle diameter).


Mobile phase: A mixture of ammonium acetate TS, 1,4-dioxane, tetrahydrofuran, water and ammonia solution (28)(100:75:50:23:2).

Flow rate: Adjust the ‰ow rate so that the retention timeof tuberactinomycin N is about 9 minutes.System suitability—

System performance: When the procedure is run with 3 mLof the sample solution under the above operating conditions,tuberactinomycin N and tuberactinomycin O are eluted inthis order with the resolution between these peaks being notless than 1.5.

System repeatability: When the test is repeated 6 timeswith 3 mL of the sample solution under the above operatingconditions, the relative standard deviation of the peak areaof tuberactinomycin N is not more than 2.0z.

Purity (1) Clarity and color of solution—Dissolve 1.0 gof Enviomycin Sulfate in 10 mL of water: the solution isclear and colorless.

(2) Heavy metals—Proceed with 2.0 g of EnviomycinSulfate according to Method 1, and perform the test. Pre-pare the control solution with 2.0 mL of Standard LeadSolution (not more than 10 ppm).

(3) Arsenic—Prepare the test solution with 2.0 g ofEnviomycin Sulfate according to Method 1, and perform thetest (not more than 1 ppm).

Loss on drying Not more than 4.0z (0.2 g, in vacuum,phosphorus (V) oxide, 609C, 3 hours).



(1) Test organism—Bacillius subtilis ATCC 6633(2) Culture medium—Use the medium i in 1) Medium


(3) Standard solutions—Weigh accurately an amount ofEnviomycin Sulfate Reference Standard, equivalent toabout 20 mg (potency), dissolve in water to make exactly20 mL, and use this solution as the standard stock solution.Keep the standard stock solution at a temperature not ex-ceeding 59C and use within 10 days. Take exactly a suitableamount of the standard stock solution before use, add0.1 mol/L phosphate buŠer solution, pH 8.0 to make solu-tions so that each mL contains 400 mg (potency) and 100 mg(potency), and use these solutions as the high concentrationstandard solution and the low concentration standard solu-tion, respectively.

(4) Sample solutions—Weigh accurately an amount ofEnviomycin Sulfate, equivalent to about 20 mg (potency),and dissolve in water to make exactly 20 mL. Take exactly asuitable amount of this solution, add 0.1 mol/L phosphatebuŠer solution, pH 8.0 to make solutions so that each mLcontains 400 mg (potency) and 100 mg (potency), and usethese solutions as the high concentration sample solutionand the low concentration sample solution, respectively.


Add the following:

Epirubicin Hydrochloride塩酸エピルビシン

C27H29NO11.HCl: 579.98(2S,4S )-4-(3-Amino-2,3,6-trideoxy-a-L-arabino-hexopyranosyloxy)-1,2,3,4-tetrahydro-2,5,12-trihydroxy-2-hydroxyacetylnaphthacene-6,11-dionemonohydrochloride [56390-09-1]

Epirubicin Hydrochloride contains not less than900 mg (potency) per mg, calculated on the anhydrousbasis and corrected by the amount of the residual sol-vent. The potency of Epirubicin Hydrochloride is ex-pressed as mass (potency) of epirubicin hydrochloride(C27H29NO11.HCl).

Description Epirubicin Hydrochloride occurs as a pale yel-lowish red to brownish red powder.

It is soluble in water and in methanol, slightly soluble in

ethanol (95), and practically insoluble in acetonitrile.It is hygroscopic.

Identiˆcation (1) Determine the absorption spectrum ofa solution of Epirubicin Hydrochloride in methanol (3 in200,000) as directed under the Ultraviolet-visible Spec-trophotometry, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities ofabsorption at the same wavelengths.

(2) Determine the infrared absorption spectrum ofEpirubicin Hydrochloride and Epirubicin HydrochlorideReference Standard as directed in the potassium bromidedisk method under the Infrared Spectrophotometry, andcompare these spectra: both spectra exhibit similar intensi-ties of absorption at the same wave numbers.


the anhydrous basis and collected by the amount of theresidual solvent, methanol, 20 mL, 100 mm).

pH Dissolve 10 mg of Epirubicin Hydrochloride in 2 mLof water: the pH of the solution is between 4.0 and 5.5.

Purity (1) Clarity and color of solution－Dissolve 50 mgof Epirubicin Hydrochloride in 5 mL of water: the solutionis clear and dark red.

(2) Heavy metals—Proceed with 1.0 g of EpirubicinHydrochloride according to Method 2, and perform the test.Prepare the control solution with 2.0 mL of Standard LeadSolution (not more than 20 ppm).

(3) Related substances—Perform the test with 10 mL ofthe sample solution obtained in the Assay as directed underthe Liquid Chromatography according to the following con-ditions, determine each peak area by the automatic integra-tion method, and calculate the sum amount of the peaksother than epirubicin and 2-naphthalenesulfonic acid by thearea percentage method: not more than 5.0z.Operating conditions—


Time span of measurement: About 3 times as long as theretention time of epirubicin after the solvent peak.System suitability—

Test for required detectability: To 1 mL of the samplesolution add the mobile phase to make 100 mL, and use thissolution as the solution for system suitability test. Pipet1 mL of the solution for system suitability test, and add themobile phase to make exactly 10 mL. Conˆrm that the peakarea of epirubicin obtained from 10 mL of this solution isequivalent to 7 to 13z of that obtained from 10 mL of thesolution for system suitability test.


(4) Residual solvents—Weigh accurately about 0.3 g ofEpirubicin Hydrochloride, add exactly 0.6 mL of the inter-nal standard solution, add N,N-dimethylformamide tomake 6 mL, and use this solution as the sample solution.Separately, pipet 1 mL of methanol, add N,N-dimethylfor-mamide to make exactly 25 mL, and use this solution as


methanol standard stock solution. Take exactly 125 mL ofacetone, 30 mL of ethanol (99.5), 32 mL of 1-propanol and17 mL of the methanol standard stock solution, add exactly10 mL of the internal standard solution and N,N-dimethyl-formamide to make 100 mL, and use this solution as thestandard solution. Perform the test with 1 mL each of thesample solution and the standard solution as directed underthe Gas Chromatography according to the following condi-tion, and determine the ratios of the peak areas of acetone,ethanol, 1-propanol and methanol to that of the internalstandard, QTA and QSA, QTB and QSB, QTC and QSC, and QTD

and QSD, respectively. Calculate the amounts of acetone,ethanol, 1-propanol and methanol by the following equa-tions: the amounts of acetone, ethanol, 1-propanol andmethanol are not more than 1.5z, not more than 0.5z, notmore than 0.5z and not more than 0.1z, respectively.

Amount (z) of acetone＝ 1WT×

QTA

QSA× 593

Amount (z) of ethanol＝1

WT×

QTB

QSB× 142

Amount (z) of 1-propanol＝1

WT×

QTC

QSC× 154

Amount (z) of methanol＝1

WT×

QTD

QSD× 2.23

WT: Amount (mg) of Epirubicin Hydrochloride

Internal standard solution—A solution of 1,4-dioxane inN,N-dimethylformamide (1 in 100).Operating conditions—

Detector: Hydrogen ‰ame-ionization detector.Column: A fused silica column 0.53 mm in inside di-

ameter and 30 m in length, coated with polyethylene glycolfor gas-chromatography 1 mm in thickness.

Column temperature: 409C for 11 minutes after injectionof the sample, then rise to 909C at a rate of 109C per minute.If necessary, rise to 1309C at a rate of 509C per minute andmaintain the temperature for 30 minutes.

Injection port temperature: A constant temperature ofabout 1209C.

Detector temperature: A constant temperature of about1509C.

Carrier gas: HeriumFlow rate: Adjust the ‰ow rate so that the retention time

of the internal standard is about 8 minutes.Split ratio: 1:15


of the standard solution under the above operating condi-tions, acetone, methanol, ethanol, 1-propanol and the inter-nal standard are eluted in this order with the resolution be-tween the peaks of acetone and the internal standard beingnot less than 30.

System repeatability: When the test is repeated 6 timeswith 1 mL of the standard solution under the above operat-ing conditions, the relative standard deviations of the peakareas of acetone, methanol, ethanol and 1-propanol are notmore than 4.0z, respectively.



Assay Weigh accurately an amount of Epirubicin Hydro-chloride and Epirubicin Hydrochloride Reference Standard,equivalent to about 50 mg (potency), dissolve each in theinternal standard solution to make exactly 50 mL, and usethese solutions as the sample solution and the standard solu-tion, respectively. Perform the test with 10 mL each of thesample solution and the standard solution as directed underthe Liquid Chromatography according to the following con-ditions, and determine the ratios, QT and QS, of the peakarea of epirubicin to that of the internal standard.


＝WS×QT

QS× 1000

WS: Amount [mg (potency)] of Epirubicin HydrochlorideReference Standard

Internal standard solution—A solution of sodium 2-naphthalene sulfonate in a mixture of water, acetonitrile,methanol and phosphoric acid (540:290:170:1) (1 in 2000).Operating conditions—


Column: A stainless steel column 4 mm in inside diameterand 25 cm in length, packed with trimethylsilanized silica gelfor liquid chromatography (6 mm in particle diameter).


Mobile phase: Dissolve 2 g of sodium lauryl sulfate in amixture of water, acetonitrile, methanol and phosphoricacid (540:290:170:1) to make 1000 mL.

Flow rate: Adjust the ‰ow rate so that the retention timeof epirubicin is about 9.5 minutes.System suitability—

System performance: When the procedure is run with10 mL of the standard solution under the above operatingconditions, the internal standard and epirubicin are eluted inthis order with the resolution between these peaks being notless than 20.

System repeatability: When the test is repeated 5 timeswith 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratiosof the peak area of epirubicin to that of the internal standardis not more than 1.0z.

Containers and storage Containers—Tight containers.Storage—At a temperature between 09C and 59C.


Ergocalciferolエルゴカルシフェロール


Assay Weigh accurately about 30 mg each of Ergocal-ciferol and Ergocalciferol Reference Standard, and dissolveeach in isooctane to make exactly 50 mL. Pipet 10 mL eachof these solutions, add exactly 3 mL each of the internalstandard solution, then add the mobile phase to make50 mL, and use these solutions as the sample solution andthe standard solution, respectively. Perform the test with 10to 20 mL each of the sample solution and the standard solu-tion as directed under the Liquid Chromatography accord-ing to the following conditions, and calculate the ratios, QT

and QS, of the peak area of ergocalciferol to that of theinternal standard. Perform the procedure rapidly avoidingcontact with air or other oxidizing agents and using light-resistant containers.

Amount (mg) of C28H44O＝WS×QT

QS

WS: Amount (mg) of Ergocalciferol Reference Standard

Internal standard solution—A solution of dimethyl phtha-late in isooctane (1 in 100).Operating conditions—


Column: A stainless steel column 4.6 mm in insidediameter and 25 cm in length, packed with a silica gel for liq-uid chromatography (10 mm particle diameter).


Mobile phase: A mixture of hexane and n-amylalcohol(997:3).

Flow rate: Adjust the ‰ow rate so that the retention timeof ergocalciferol is about 25 minutes.System suitability—

System performance: Dissolve 15 mg of ErgocalciferolReference Standard in 25 mL of isooctane. Transfer thissolution to a ‰ask, heat in an oil bath under a re‰ux con-denser for 2 hours, and cool immediately to room tempera-ture. Transfer the solution to a quartz test tube, and irradi-ate with a short-wave lamp (main wavelength: 254 nm) and along-wave lamp (main wavelength: 365 nm) for 3 hours. To10 mL of this solution add the mobile phase to make 50 mL.When the procedure is run with 10 mL of this solution underthe above operating conditions. Use a column with the ratiosof the retention time of previtamin D2, trans-vitamin D2 andtachysterol2 to that of ergocalciferol being about 0.5, about0.6 and about 1.1, respectively, and with resolution betweenprevitamin D2 and trans-vitamin D2 being not less than 0.7,and that between ergocalciferol and tachysterol2 being notless than 1.0.

System repeatability: When the test is repeated 6 timeswith 10 mL of the standard solution under the above operat-

ing conditions, the relative standard deviation of the ratiosof the peak area of ergocalciferol to that of the internal stan-dard is not more than 1.0z.

Erythromycinエリスロマイシン


Erythromycin contains not less than 930 mg(potency) per mg, calculated on the anhydrous basis.The potency of Erythromycin is expressed as mass(potency) of erythromycin (C37H67NO13).

Description Erythromycin occurs as a white to light yel-lowish white powder.

It is very soluble in N,N-dimethylformamide, freely solu-ble in methanol and in ethanol (95), and very slightly solublein water.

Identiˆcation (1) Determine the infrared absorptionspectrum of Erythromycin as directed in the potassiumbromide disk method under the Infrared Spectrophotomet-ry, and compare the spectrum with the Reference Spectrumor the spectrum of Erythromycin Reference Standard: bothspectra exhibit similar intensities of absorption at the samewave numbers.

(2) Dissolve 0.01 g each of Erythromycin andErythromycin Reference Standard in 1 mL of methanol, anduse these solutions as the sample solution and the standardsolution. Perform the test with these solutions as directedunder the Thin-layer Chromatography. Spot 5 mL each ofthe sample solution and the standard solution on a plate ofsilica gel for thin-layer chromatography. Develop the platewith a mixture of methanol and ammonia solution (28)(50:1) to a distance of about 10 cm, and air-dry the plate.Spray evenly 4-methoxybenzaldehyde-sulfuric acid TS onthe plate, and heat at 1009C for 15 minutes: the principalspot from the sample solution and the spot from the stan-dard solution are dark purple in color, and their R f valuesare the same.

Optical rotation [a]20D : －71 –－789(1 g calculated on the

anhydrous basis, ethanol (95), 50 mL, 100 mm).

Purity (1) Heavy metals—Proceed with 1.0 g ofErythromycin according to Method 4, and perform the test.Prepare the control solution with 2.0 mL of Standard LeadSolution (not more than 20 ppm).

(2) Arsenic—Prepare the test solution with 1.0 g ofErythromycin according to Method 5 using hydrochloricacid instead of diluted hydrochloric acid (1 in 2), and per-form the test (not more than 2 ppm).

(3) Related substances—Dissolve 40 mg of Erythromy-cin in 2 mL of methanol, add a mixture of phosphate buŠersolution, pH 7.0 and methanol (15:1) to make exactly10 mL, and use this solution as the sample solution.


Separately, dissolve 16 mg of Erythromycin Reference Stan-dard in 2 mL of methanol, add a mixture of phosphatebuŠer solution, pH 7.0 and methanol (15:1) to make exactly10 mL, and use this solution as the standard stock solution.Dissolve 5 mg each of erythromycin B and erythromycin Cin 2 mL of methanol, add exactly 2 mL of the standard stocksolution, add a mixture of phosphate buŠer solution, pH 7.0and methanol (15:1) to make exactly 25 mL, and use thissolution as the standard solution. Perform the test with ex-actly 100 mL each of the sample solution and the standardsolution as directed under the Liquid Chromatographyaccording to the following conditions, and determine eachpeak area by the automatic integration method: the peakareas of erythromycin B and erythromycin C from the sam-ple solution are not more than those of erythromycin B anderythromycin C from the standard solution, respectively,and each area of the peaks other than erythromycin,erythromycin B and erythromycin C is not more than thearea of the peak of erythromycin from the standard solu-tion.Operating conditions—




Mobile phase: Dissolve 3.5 g of dipotassium hydrogenphosphate in water to make 100 mL, and adjust the pH to9.0 with diluted phosphoric acid (1 in 10). To 50 mL of thissolution add 190 mL of t-butanol, 30 mL of acetonitrile andwater to make 1000 mL.

Flow rate: Adjust the ‰ow rate so that the retention timeof erythromycin is about 20 minutes.

Time span of measurement: About 4 times as long as theretention time of erythromycin after the solvent peak.System suitability—

System performance: Dissolve 2 mg of N-demethyl-erythromycin in 10 mL of the standard solution. When theprocedure is run with 100 mL of this solution under theabove operating conditions, N-demethylerythromycin,erythromycin C, erythromycin and erythromycin B are elut-ed in this order, with the resolution between the peaks of N-demethylerythromycin and erythromycin C being not lessthan 0.8, and with the resolution between the peaks of N-demethylerythromycin and erythromycin being not less than5.5.

System repeatability: When the test is repeated 3 timeswith 100 mL of the standard solution under the above oper-ating conditions, the relative standard deviation of the peakarea of erythromycin is not more than 3.0z.





(2) Culture medium—Use the medium i in 3) Mediumfor other organisms under (1) Agar media for seed and baselayer. Adjust the pH of the medium so that it will be 7.8 to8.0 after sterilization.

(3) Standard solutions—Weigh accurately an amount ofErythromycin Reference Standard, equivalent to about25 mg (potency), dissolve in 25 mL of methanol, add0.1 mol/L phosphate buŠer solution, pH 8.0 to makeexactly 100 mL, and use this solution as the standard stocksolution. Keep the standard stock solution at 59C or below,and use within 7 days. Take exactly a suitable amount of thestandard stock solution before use, add 0.1 mol/L phos-phate buŠer solution, pH 8.0 to make solutions so that eachmL contains 20 mg (potency) and 5 mg (potency), and usethese solutions as the high concentration standard solutionand the low concentration standard solution, respectively.

(4) Sample solutions—Weigh accurately an amount ofErythromycin, equivalent to about 25 mg (potency), dissolvein 25 mL of methanol, and add 0.1 mol/L phosphate buŠersolution, pH 8.0 to make exactly 100 mL. Take exactly asuitable amount of this solution, add 0.1 mol/L phosphatebuŠer solution, pH 8.0 to make solutions so that each mLcontains 20 mg (potency) and 5 mg (potency), and use thesesolutions as the high concentration sample solution and thelow concentration sample solution, respectively.

Containers and storage Containers—Well-closed contain-ers.


Add the following:

Erythromycin Lactobionateラクトビオン酸エリスロマイシン

C37H67NO13.C12H22O12: 1092.22(2R,3S,4S,5R,6R,8R,10R,11R,12S,13R )-5-(3,4,6-Trideoxy-3-dimethylamino-b-D-xylo-hexopyranosyloxy)-3-(2,6-dideoxy-3-C-methyl-3-O-methyl-a-L-ribo-hexopyranosyloxy)-6,11,12-trihydroxy-2,4,6,8,10,12-hexamethyl-9-oxopentadecan-13-olide mono(4-O-b-D-galactopyranosyl-D-gluconate) [3847-29-8]

Erythromycin Lactobionate contains not less than590 mg (potency) per mg, calculated on the anhydrousbasis. The potency of Erythromycin Lactobionateis expressed as mass (potency) of erythromycin(C37H67NO13: 733.93).

Description Erythromycin Lactobionate occurs as a whitepowder.

It is freely soluble in water, in methanol and in ethanol(99.5), and very slightly soluble in acetone.

Identiˆcation (1) To 3 mg of Erythromycin Lactobionateadd 2 mL of acetone, and add 2 mL of hydrochloric acid: anorange color is produced, and it changes immediately to redto deep purple.

(2) Transfer about 0.3 g of Erythromycin Lactobionateto a separator, add 15 mL of ammonia TS and 15 mL ofchloroform, shake, and take the separated aqueous layer.Wash the aqueous layer with three 15-mL portions of chlo-roform, and evaporate the aqueous liquid on a water bath todryness. Dissolve the residue in 10 mL of a mixture ofmethanol and water (3:2), and use this solution as the samplesolution. Separately, dissolve 0.10 g of lactobionic acid in10 mL of a mixture of methanol and water (3:2), and use thissolution as the standard solution. Perform the test with thesesolutions as directed under the Thin-layer Chromatography.Spot 10 mL each of the sample solution and the standardsolution on a plate of silica gel for thin-layer chromato-graph. Develop the plate with the upper layer obtained froma mixture of water, 1-butanol and acetic acid (100) (3:3:1) toa distance of about 10 cm, and air-dry the plate. Sprayevenly dilute sulfuric acid, and heat at 1059C for 20 minutes:the principal spot obtained from the sample solution shows adeep brown and the Rf value which are the same as those ofthe principal spot from the standard solution.

pH The pH of a solution obtained by dissolving 0.5 g ofErythromycin Lactobionate in 10 mL of water is between 5.0and 7.5.




(2) Culture medium—Use the medium i in 3) Mediumfor other organisms under (1) Agar media for seed and baselayer. Adjust the pH of the medium so that it will be 7.8 to8.0 after sterilization.

(3) Standard solutions—Weigh accurately an amountof Erythromycin Reference Standard, equivalent to about50 mg (potency), dissolve in 50 mL of methanol, add0.1 mol/L phosphate buŠer solution, pH 8.0 to makeexactly 100 mL, and use this solution as the standard stocksolution. Keep the standard stock solution at not exceeding59C and use within 7days. Take exactly a suitable amount ofthe standard stock solution before use, add 0.1 mol/L phos-phate buŠer solution, pH 8.0 to make solutions so that eachmL contains 20 mg (potency) and 5 mg (potency), and usethese solutions as the high concentration standard solutionand the low concentration standard solution, respectively.

(4) Sample solutions—Weigh accurately an amount ofErythromycin Lactobionate, equivalent to about 50 mg(potency), dissolve in 50 mL of methanol, and add0.1 mol/L phosphate buŠer solution, pH 8.0 to makeexactly 100 mL. Take exactly a suitable amount of this solu-tion, add 0.1 mol/L phosphate buŠer solution, pH 8.0 tomake solutions so that each mL contains 20 mg (potency) and5 mg (potency), and use these solutions as the high concentra-tion sample solution and the low concentration sample solu-tion, respectively.


Estradiol Benzoate安息香酸エストラジオール


Assay Weigh accurately about 10 mg each of EstradiolBenzoate and Estradiol Benzoate Reference Standard, previ-ously dried, and dissolve each in methanol to make exactly20 mL. Pipet 5 mL each of these solutions, add 5 mL of theinternal standard solution, then add methanol to make20 mL, and use these solutions as the sample solution andthe standard solution, respectively. Perform the test with5 mL of the sample solution and the standard solution asdirected under the Liquid Chromatography according to thefollowing conditions, and calculate the ratios, QT and QS, ofthe peak area of estradiol benzoate to that of the internalstandard.


Amount (mg) of C25H28O3＝WS×QT

QS

WS: Amount (mg) of Estradiol Benzoate ReferenceStandard

Internal standard solution—A solution of progesterone inmethanol (13 in 80,000).Operating conditions—





of estradiol benzoate is about 10 minutes.System suitability—

System performance: When the procedure is run with 5 mLof the standard solution under the above operating condi-tions, the internal standard and estradiol benzoate are elutedin this order with the resolution between these peaks beingnot less than 9.

System repeatability: When the test is repeated 6 timeswith 5 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratiosof the peak area of estradiol benzoate to that of the internalstandard is not more than 1.0z.

Estriolエストリオール


Assay Weigh accurately about 25 mg each of Estriol andEstriol Reference Standard, previously dried, and dissolveeach in methanol to make exactly 50 mL. Pipet 10 mL eachof these solutions, add exactly 5 mL of the internal standardsolution, add methanol to make 100 mL, and use these solu-tions as the sample solution and the standard solution,respectively. Perform the test with 10 mL each of the samplesolution and the standard solution as directed under the Liq-uid Chromatography according to the following conditions,and calculate the ratios, QT and QS, of the peak area ofestriol to that of the internal standard, respectively.


QS

WS: Amount (mg) of Estriol Reference Standard

Internal standard solution—A solution of methyl benzoatefor estriol limit test in methanol (1 in 1000).Operating conditions—





of estriol is about 10 minutes.System suitability—

System performance: When the procedure is run with10 mL of the standard solution under the above operatingconditions, estriol and the internal standard are eluted in thisorder with the resolution between these peaks being not lessthan 8.

System repeatability: When the test is repeated 6 timeswith 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratiosof the peak area of estriol to that of the internal standard isnot more than 1.0z.

Add the following:

Etizolamエチゾラム

C17H15ClN4S: 342.854-(2-Chlorophenyl)-2-ethyl-9-methyl-6H-thieno[3,2-f ][1,2,4]triazolo[4,3-a][1,4]diazepine[40054-69-1]

Etizolam contains not less than 98.5z and not morethan 101.0z of etizolam (C17H15ClN4S).

Description Etizolam occurs as a white to pale yellowishwhite crystalline powder.

It is soluble in ethanol (99.5), sparingly soluble in aceto-nitrile and in acetic anhydride, and practically insoluble inwater.

Identiˆcation (1) Determine the absorption spectrum ofa solution of Etizolam in ethanol (99.5) (1 in 100,000) asdirected under the Ultraviolet-visible Spectrophotometry,and compare the spectrum with the Reference Spectrum:both spectra exhibit similar intensities of absorption at thesame wavelengths.

(2) Determine the infrared absorption spectrum ofEtizolam as directed in the potassium bromide disk methodunder the Infrared Spectrophotometry, and compare thespectrum with the Reference Spectrum: both spectra exhibit


similar intensities of absorption at the same wave numbers.


Purity (1) Heavy metals—Proceed with 2.0 g of Etizolamaccording to Method 2, and perform the test. Prepare thecontrol solution with 2.0 mL of Standard Lead Solution (notmore than 10 ppm).

(2) Related substances—Dissolve 20 mg of Etizolam in50 mL of acetonitrile, and use this solution as the samplesolution. Pipet 1 mL of the sample solution, and addacetonitrile to make exactly 20 mL. Pipet 1 mL of this solu-tion, add acetonitrile to make exactly 50 mL, and use thissolution as the standard solution. Perform the test withexactly 10 mL each of the sample solution and the standardsolution as directed under the Liquid Chromatographyaccording to the following conditions, and determine eachpeak area by the automatic integration method: the area ofthe peak other than etizolam obtained from the sample solu-tion is not more than the peak area of etizolam from thestandard solution.Operating conditions—




Mobile phase: Dissolve 1.36 g of potassium dihydrogenphosphate in water to make 1000 mL, and adjust the pH to3.5 with potassium hydroxide TS. To 550 mL of this solu-tion add 450 mL of acetonitrile.

Flow rate: Adjust the ‰ow rate so that the retention timeof etizolam is about 6 minutes.

Time span of measurement: About 5 times as long as theretention time of etizolam after the solvent peak.System suitability—

Test for required detectability: Measure exactly 2 mL ofthe standard solution, and add acetonitrile to make exactly20 mL. Conˆrm that the peak area of etizolam obtainedfrom 10 mL of this solution is equivalent to 8 to 12z of thatfrom 10 mL of the standard solution.

System performance: Dissolve 0.02 g each of Etizolamand ethyl parahydroxybenzoate in the mobile phase to make50 mL. To 1 mL of this solution add the mobile phase tomake 50 mL. When the procedure is run with 10 mL of thissolution under the above operating conditions, etizolam andethyl parahydroxybenzoate are eluted in this order with theresolution between these peaks being not less than 3.

System repeatability: When the test is repeated 6 timeswith 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peakarea of etizolam is not more than 2z.



Assay Weigh accurately about 0.3 g of Etizolam, previous-ly dried, dissolve in 70 mL of a mixture of acetic anhydrideand acetic acid (100) (7:3), and titrate with 0.1 mol/L per-chloric acid VS (potentiometric titration). The end point isthe second equivalent point. Perform a blank determination,and make any necessary correction.

Each mL of 0.1 mol/L perchloric acid VS＝ 17.14 mg of C17H15ClN4S


Famotidine for Injection注射用ファモチジン


Purity(2) Related substances—Take a number of Famotidine

for Injection, equivalent to about 0.1 g of famotidine(C8H15N7O2S3), dissolve each content in water, wash theinside of the container with water, combine the solutions ofthe contents with the washings, add water to the combinedsolution to make exactly 100 mL, and use this solution as thesample solition. Pipet 1 mL of the sample solution, addwater to make exactly 100 mL, and use this solution as thestandard solution. Perform the test with exactly 5 mL each ofthe sample solution and the standard solution as directed un-der the Liquid Chromatography according to the followingconditions, and determine each peak area of these solutionsby the automatic integration method: the total area of thepeaks other than peak of famotidine from the sample solu-tion is not larger than peak area of famotidine from the stan-dard solution.Operating conditions—


Time span of measurement: About 2 times as long as theretention time of famotidine after the solvent peak.System suitability—

Test for required detection: To exactly 2 mL of the stan-dard solution add the water to make exactly 20 mL. Conˆrmthat the peak area of famotidine obtained from 5 mL of thissolution is equivalent to 8 to 12z of that of famotidineobtained from 5 mL of the standard solution.


System repeatability: When the test is repeated 6 timeswith 5 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peakarea of famotidine is not more than 2.0z.


Delete the following Monograph:

Floctafenineフロクタフェニン

Flomoxef Sodiumフロモキセフナトリウム


Flomoxef Sodium contains not less than 862 mg(potency) per mg, calculated on the anhydrous basis.The potency of Flomoxef Sodium is expressed as mass(potency) of ‰omoxef (C15H18F2N6O7S2: 496.47).

Description Flomoxef Sodium occurs as white to light yel-lowish white, powder or masses.

It is very soluble in water, freely soluble in methanol, andsparingly soluble in ethanol (99.5).

Identiˆcation (1) Decompose 0.01 g of Flomoxef Sodiumas directed under the Oxygen Flask Combustion Method,using a mixture of 0.5 mL of 0.01 mol/L sodium hydroxideTS and 20 mL of water as the absorbing liquid. To 2 mL ofthe test solution so obtained add 1.5 mL of a mixture ofalizarin complexone TS, acetic acid-potassium acetate buŠersolution, pH 4.3 and cerium (III) nitrate TS (1:1:1): blue-purple color develops.

(2) Determine the absorption spectrum of a solution ofFlomoxef Sodium (3 in 100,000) as directed under theUltraviolet-visible Spectrophotometry, and compare thespectrum with the Reference Spectrum: both spectra exhibitsimilar intensities of absorption at the same wavelengths.

(3) Determine the infrared absorption spectrum ofFlomoxef Sodium as directed in the potassium bromide diskmethod under the Infrared Spectrophotometry, and com-pare the spectrum with the Reference Spectrum: both spec-tra exhibit similar intensities of absorption at the same wavenumbers.

(4) Determine the spectrum of a solution of FlomoxefSodium in heavy water for nuclear magnetic resonance spec-troscopy (1 in 100) as directed under the Nuclear MagneticResonance Spectroscopy (1H), using sodium 3-trimethylsilyl-propanesulfonate for nuclear magnetic resonance spec-troscopy as an internal reference compound: it exhibits a sin-gle signal A at around d3.5 ppm, a single signal or a sharpmultiple signal B at around d3.7 ppm, and a single signal Cat around d5.2 ppm. The ratio of the integrated intensity ofthese signals, A:B:C, is about 3:2:1.

(5) Flomoxef Sodium responds to the Qualitative Test(1) for sodium salt.

Optical rotation [a]20D : －8 –－139(1 g calculated on the

anhydrous basis, a mixture of water and ethanol (99.5) (4:1),

50 mL, 100 mm).

pH The pH of a solution obtained by dissolving 0.5 g ofFlomoxef Sodium in 5 mL of water is between 4.0 and 5.5.

Purity (1) Clarity and color of solution—Dissolve 1.0 gof Flomoxef Sodium in 10 mL of water: the solution is clearand pale yellow.

(2) Heavy metals—Proceed with 1.0 g of FlomoxefSodium in a quartz crucible according to Method 2, andperform the test. Prepare the control solution with 2.0 mLof Standard Lead Solution (not more than 20 ppm).

(3) Arsenic—To 1.0 g of Flomoxef Sodium 5 mL of sul-furic acid and 5 mL of nitric acid, heat carefully until thesolution changes to colorless to light yellow with occasionaladdition of 2 mL of nitric acid. After cooling, add 10 mL ofammonium oxalate TS, heat until white fumes evolve, andconcentrate to 2 to 3 mL. After cooling, add water to make10 mL, and perform the test using this solution as the testsolution: the color is not darker than that of the controlsolution.

Control solution: Proceed to prepare a solution in thesame manner as the test solution without Flomoxef Sodium,and transfer 10 mL of the solution so obtained to the genera-tor bottle, add exactly 2 mL of Standard Arsenic Solution,and proceed in the same manner as the test solution (notmore than 2 ppm).

(4) 1-(2-Hydroxyethyl)-1H-tetrazol-5-thiol—Weigh ac-curately about 20 mg of 1-(2-hydroxyethyl)-1H-tetrazol-5-thiol, and dissolve in water to make exactly 100 mL. Pipet5 mL of this solution, add exactly 25 mL of the internal stan-dard solution and water to make 50 mL, and use this solu-tion as the standard solution. Perform the test with 5 mLeach of the sample solution obtained in the Assay and thestandard solution as directed under the Liquid Chro-matography according to the following conditions, anddetermine the ratios, QT and QS, of the peak area of 1-(2-hydroxyethyl)-1H-tetrazol-5-thiol to that of the internalstandard: the amount of 1-(2-hydroxyethyl)-1H-tetrazol-5-thiol is not more than 1.0z of the amount of FlomoxefSodium calculated on the anhydrous basis.

Amount (mg) of 1-(2-hydroxyethyl)-1H-tetrazol-5-thiol(C3H6N4OS)

＝WS×QT

QS×

110

WS: Amount (mg) of 1-(2-hydroxyethyl)-1H-tetrazol-5-thiol

Operating conditions—Proceed as directed in the operating conditions in the

Assay.System suitability—

Proceed as directed in the system suitability in the Assay.


Assay Weigh accurately an amount of Flomoxef Sodiumand Flomoxef Triethylammonium Reference Standard,


equivalent to about 50 mg (potency), and dissolve each in ex-actly 50 mL of the internal standard solution, add water tomake 100 mL, and use these solutions as the sample solutionand standard solution. Perform the test with 5 mL each ofthe sample solution and the standard solution as directed un-der the Liquid Chromatography according to the followingconditions, and determine the ratios, QT and QS, of the peakarea of ‰omoxef to that of the internal standard.

Amount [mg (potency)] of ‰omoxef (C15H18F2N6O7S2)

＝WS×QT

QS× 1000

WS: Amount [mg (potency)] of Flomoxef Triethylammo-nium Reference Standard

Internal standard solution—A solution of m-cresol (3 in1000).Operating conditions—


Column: A stainless steel column 4 mm in inside diameterand 20 cm in length, packed with octadecylsilanized silica gelfor liquid chromatography (5 – 10 mm in particle diameter).


Mobile phase: Dissolve 6.94 g of potassium dihydrogenphosphate, 3.22 g of disodium hydrogen phosphate 12-waterand 1.60 g of tetra-n-butylammonium bromide in water tomake 1000 mL. To 750 mL of this solution add 250 mL ofmethanol.

Flow rate: Adjust the ‰ow rate so that the retention timeof ‰omoxef is about 9 minutes.System suitability—

System performance: When the procedure is run with 5 mLof the standard solution under the above operating condi-tions, ‰omoxef and the internal standard are eluted in thisorder with the resolution between these peaks being not lessthan 10.

System repeatability: When the test is repeated 3 timeswith 5 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratiosof the peak area of ‰omoxef to that of the internal standardis not more than 1.0z.

Containers and storage Containers—Tight containers.Storage—At 59C or below.

Flurbiprofenフルルビプロフェン


Purity(3) Related substances—Dissolve 20 mg of Flurbiprofen

in 10 mL of a mixture of water and acetonitrile (11:9), anduse this solution as the sample solution. Pipet 1 mL of thesample solution, and add a mixture of water and acetonitrile

(11:9) to make exactly 200 mL, and use this solution as thestandard solution. Perform the test with exactly 20 mL eachof the sample solution and the standard solution as directedunder the Liquid Chromatography according to the follow-ing conditions. Determine each peak area of both solutionsby the automatic integration method: each area of the peaksother than the peak of ‰urbiprofen from the sample solutionis not larger than the peak area of ‰urbiprofen from thestandard solution, and the total area of these peaks is notlarger than twice the peak area of ‰urbiprofen from the stan-dard solution.Operating conditions—





Flow rate: Adjust the ‰ow rate so that the retention timeof ‰urbiprofen is about 20 minutes.

Time span of measurement: About twice as long as theretention time of ‰urbiprofen after the solvent peak.System suitability—

Test for required detection: To exactly 5 mL of the stan-dard solution add a mixture of water and acetonitrile (11:9)to make exactly 25 mL. Conˆrm that the peak area of ‰ur-biprofen obtained from 20 mL of this solution is equivalentto 16 to 24z of that of ‰urbiprofen obtained from 20 mL ofthe standard solution.

System performance: Dissolve 0.04 g of ‰urbiprofen and0.02 g of butyl parahydroxybenzoate in 100 mL of a mixtureof water and acetonitrile (11:9). To 5 mL of this solution adda mixture of water and acetonitrile (11:9) to make 50 mL.When the procedure is run with 20 mL of this solution underthe above operating conditions, butyl parahydroxybenzoateand ‰urbiprofen are eluted in this order with the resolutionbetween these peaks being not less than 12.

System repeatability: When the test is repeated 6 timeswith 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peakarea of ‰urbiprofen is not more than 2.0z.

Folic Acid葉酸


Water Not more than 8.5z (10 mg, coulometric titration).


Fradiomycin Sulfate硫酸フラジオマイシン


Fradiomycin Sulfate, when dried, contains not lessthan 623 mg (potency) per mg. The potency ofFradiomycin Sulfate is expressed as mass (potency) offradiomycin (C23H46N6O13: 614.64).

Description Fradiomycin Sulfate occurs as a white to lightyellow powder.

It is freely soluble in water, and practically insoluble inethanol (95).

It is hygroscopic.

Identiˆcation (1) Dissolve 0.05 g each of FradiomycinSulfate and Fradiomycin Sulfate Reference Standard in1 mL of water, and use these solutions as the sample solutionand the standard solution. Perform the test with these solu-tions as directed under the Thin-layer Chromatography.Spot 2 mL each of the sample solution and the standard solu-tion on a plate of silica gel for thin-layer chromatography.Develop the plate with a mixture of methanol, ammoniasolution (28) and dichloromethane (3:2:1) to a distance ofabout 10 cm, and air-dry the plate. Spray evenly a solutionof ninhydrin in acetone (1 in 50) on the plate, and heat at1109C for 15 minutes: the Rf values of the principal spotsfrom the sample solution and the standard solution are notdiŠerent each other.

(2) A solution of Fradiomycin Sulfate (1 in 20) respondsto the Qualitative Test (1) for sulfate.

Optical rotation [a]20D : ＋53.5 –＋59.09(1 g calculated on


pH The pH of a solution obtained by dissolving 1.0 g ofFradiomycin Sulfate in 10 mL of water is between 5.0 and7.5.

Purity (1) Heavy metals—Proceed with 1.0 g of Fra-diomycin Sulfate according to Method 2, and performthe test. Prepare the control solution with 2.0 mL of Stan-dard Lead Solution (not more than 20 ppm).

(2) Arsenic—Prepare the test solution with 1.0 g ofFradiomycin Sulfate according to Method 1, and performthe test (not more than 2 ppm).

(3) Related substances—Dissolve 0.63 g of FradiomycinSulfate in 5 mL of water, and use this solution as the samplesolution. Pipet 1 mL of the sample solution, add water tomake exactly 50 mL, and use this solution as the standardsolution. Perform the test with these solutions as directedunder the Thin-layer Chromatography. Spot 1 mL each ofthe sample solution and the standard solution on a plate ofsilica gel for thin-layer chromatography. Develop the platewith a mixture of methanol, ammonia solution (28) anddichloromethane (3:2:1) to a distance of about 10 cm, andair-dry the plate. Spray evenly a solution of ninhydrin in ace-

tone (1 in 50) on the plate, and heat at 1109C for 15 minutes:the spot at around R f 0.4 from the sample solution is notmore intense than the spot from the standard solution.

Loss on drying Not more than 8.0z (0.2 g, in vaccum,609C, 3 hours).


Assay Perform the test according to the Cylinder-platemethod as directed under the Microbial Assay for Antibio-tics according to the following conditions.


(2) Agar medium for seed and base layerGlucose 1.0 gPeptone 6.0 gMeat extract 1.5 gYeast extract 3.0 gSodium chloride 2.5 gAgar 15.0 gWater 1000 mL

Mix all the ingredients and sterilize. Adjust the pH aftersterilization to 7.8 – 8.0 with sodium hydroxide TS.

(3) Standard solutions－Weigh accurately an amount ofFradiomycin Sulfate Reference Standard, previously dried,equivalent to about 50 mg (potency), dissolve in 0.1 mol/Lphosphate buŠer solution for antibiotics, pH 8.0 to make ex-actly 50 mL, and use this solution as the standard stock solu-tion. Keep the standard stock solution at 59C or below anduse within 14 days. Take exactly a suitable amount of thestandard stock solution before use, add 0.1 mol/L phos-phate buŠer solution for antibiotics, pH 8.0 to make solu-tions so that each mL contains 80 mg (potency) and 20 mg(potency), and use these solutions as the high concentrationstandard solution and the low concentration standard solu-tion, respectively.

(4) Sample solutions—Weigh accurately an amount ofFradiomycin Sulfate, previously dried, equivalent to about50 mg (potency), dissolve in 0.1 mol/L phosphate buŠersolution for antibiotics, pH 8.0 to make exactly 50 mL. Takeexactly a suitable amount of this solution, add 0.1 mol/Lphosphate buŠer solution for antibiotics, pH 8.0 to make so-lutions so that each mL contains 80 mg (potency) and 20 mg(potency), and use these solutions as the high concentrationsample solution and the low concentration sample solution,respectively.



Gentamicin Sulfate硫酸ゲンタマイシン


Gentamicin Sulfate contains not less than 590 mg(potency) per mg, calculated on the dried basis. Thepotency of Gentamicin Sulfate is expressed as mass(potency) of gentamicin C1 (C21H43N5O7: 477.60).

Description Gentamicin Sulfate occurs as a white to lightyellowish white powder.


It is hygroscopic.

Identiˆcation (1) Dissolve 50 mg of Gentamicin Sulfatein 1 mL of water, and add 2 drops of a solution of 1-naphthol in ethanol (95) (1 in 500). Gently superimpose thissolution on 1 mL of sulfuric acid: a blue-purple color de-velops at the zone of contact.

(2) Dissolve 50 mg each of Gentamicin Sulfate and Gen-tamicin Sulfate Reference Standard in 10 mL of water, anduse these solutions as the sample solution and the standardsolution. Perform the test with these solutions as directedunder the Thin-layer Chromatography. Spot 20 mL of thesample solution and the standard solution on a plate of silicagel for thin-layer chromatography. Separately, shake a mix-ture of chloroform, ammonia solution (28) and methanol(2:1:1) in a separator, and allow the mixture to stand formore than 1 hour. To 20 mL of the lower layer so obtainedadd 0.5 mL of methanol, and use this as the developing sol-vent. Develop the plate with the developing solvent to a dis-tance of about 17 cm in a developing container with a cover,having an opening of about 20 mm2, and without putting aˆlter paper in the container, and air-dry the plate. Allow theplate to stand in iodine vapors: three principal spots ob-tained from the sample solution are the same with the corre-sponding spots from the standard solution in color tone andthe Rf value, respectively.

(3) Dissolve 50 mg of Gentamicin Sulfate in 5 mL ofwater, and add 0.5 mL of barium chloride TS: a whiteprecipitate is formed.



pH The pH of a solution obtained by dissolving 0.20 g ofGentamicin Sulfate in 5 mL of water is between 3.5 and 5.5.

Content ratio of the active principle Dissolve 50 mg ofGentamicin Sulfate in water to make 10 mL, and use thissolution as the sample solution. Perform the test with thissolution as directed under the Thin-layer Chromatography.Spot 20 mL of the sample solution on a plate of silica gel forthin-layer chromatography. Separately, shake a mixture ofchloroform, ammonia solution (28) and methanol (2:1:1) ina separator, and allow the mixture to stand for more than 1

hour. To 20 mL of the lower layer so obtained add 0.5 mLof methanol, and use this as the developing solvent. Developthe plate with the developing solvent to a distance of about17 cm in a developing container with a cover, having anopening of about 20 mm2, without putting a ˆlter paper inthe container, and air-dry the plate. Allow the plate to standin iodine vapor. Determine the integral absorbances, Aa, Ab

and Ac, of the colored spots of gentamicin C1 (R f value:about 0.3), gentamicin C2 (R f value: about 0.2) and gen-tamicin C1a (R f value: about 0.1), respectively, by a den-sitometer (wavelength: 450 nm) while covering the plate witha glass plate, and calculate these amounts by the followingformulae: gentamicin C1 is between 25z and 55z, gentami-cin C2 is between 25z and 50z, and gentamicin C1a isbetween 5z and 30z.

Amount (z) of gentamicin C1＝Aa

Aa＋ 1.35Ab＋ Ac× 100

Amount (z) of gentamicin C2＝1.35Ab

Aa＋ 1.35Ab＋ Ac× 100

Amount (z) of gentamicin C1a＝Ac

Aa＋ 1.35Ab＋ Ac× 100

Purity (1) Clarity and color of solution—Dissolve 1.0 gof Gentamicin Sulfate in 10 mL of water: the solution isclear and colorless to pale yellow.

(2) Heavy metals—Proceed with 2.0 g of GentamicinSulfate according to Method 4, and perform the test. Pre-pare the control solution with 2.0 mL of Standard LeadSolution (not more than 10 ppm).

(3) Related substances—Dissolve 50 mg of GentamicinSulfate in water to make 10 mL, and use this solution as thesample solution. Pipet 1 mL of the sample solution, addwater to make exactly 50 mL, and use this solution as thestandard solution. Perform the test with these solutions asdirected under the Thin-layer Chromatography. Spot 20 mLof the sample solution and the standard solution on a plateof silica gel for thin-layer chromatography. Separately,shake a mixture of chloroform, ammonia solution (28) andmethanol (2:1:1) in a separator, and allow the mixture tostand for more than 1 hour. To 20 mL of the lower layer soobtained add 0.5 mL of methanol, and use this as the de-veloping solvent. Develop the plate with the developing sol-vent to a distance of about 17 cm in a developing containerwith a cover, having an opening of about 20 mm2, withoutputting a ˆlter paper in the container, and air-dry the plate.Allow the plate to stand in iodine vapor, and compare thecolored spots while covering with a glass plate: the spotsother than the spots of gentamicin C1 (R f value: about 0.3),gentamicin C2 (R f value: about 0.2) and gentamicin C1a (R fvalue: about 0.1) obtained from the sample solution are notmore intense than the spot of gentamicin C2 from the stan-dard solution.

Loss on drying Not more than 18.0z (0.15 g, reducedpressure not exceeding 0.67 kPa, 1109C, 3 hours). Handlethe sample avoiding absorption of moisture.


Assay Perform the test according to the Cylinder-plate


method as directed under the Microbial Assay for Antibio-tics according to the following conditions.

(1) Test organism—Staphylococcus epidermidis ATCC12228

(2) Agar media for seed and base layer—Glucose 1.0 gPeptone 6.0 gMeat extract 1.5 gYeast extract 3.0 gSodium chloride 10.0 gAgar 15.0 gWater 1000 mL

Mix all the ingredients, and sterilize. Adjust the pH of thesolution so that it will be 7.8 to 8.0 after sterilization.

(3) Agar medium for transferring test organisms—Usethe medium ii in 2) Medium for other organisms under (2)Agar media for transferring test organisms.

(4) Standard solutions—Weigh accurately an amount ofGentamicin Sulfate Reference Standard, equivalent to about25 mg (potency), dissolve in 0.1 mol/L phosphate buŠersolution, pH 8.0 to make exactly 25 mL, and use this solu-tion as the standard stock solution. Keep the standard stocksolution at 159C or lower, and use within 30 days. Takeexactly a suitable amount of the standard stock solution be-fore use, add 0.1 mol/L phosphate buŠer solution, pH 8.0to make solutions so that each mL contains 4 mg (potency)and 1 mg (potency), and use these solutions as the high con-centration standard solution and the low concentration stan-dard solution, respectively.

(5) Sample solutions—Weigh accurately an amount ofGentamicin Sulfate, equivalent to about 25 mg (potency),and dissolve in 0.1 mol/L phosphate buŠer solution, pH 8.0to make exactly 25 mL. Take exactly a suitable amount ofthis solution, add 0.1 mol/L phosphate buŠer solution, pH8.0 to make solutions so that each mL contains 4 mg(potency) and 1 mg (potency), and use these solutions as thehigh concentration sample solution and the low concentra-tion sample solution, respectively.


Glycerinグリセリン


Glycerin contains not less than 84.0z and not morethan 87.0z of C3H8O3.


Description Glycerin is a clear, colorless, viscous liquid. Ithas a sweet taste.

It is miscible with water and with ethanol (99.5).It is hygroscopic.


Identiˆcation Determine the infrared absorption spectrumof Glycerin as directed in the liquid ˆlm method under theInfrared Spectrophotometry, and compare the spectrumwith the Reference Spectrum: both spectra exhibit similar in-tensities of absorption at the same wave numbers.

Add the following next to Purity:

Water 13 – 17z (0.1 g, volumetric titration, direct titra-tion).

Add the following next to Residue on ignition:

Assay Weigh accurately about 0.2 g of Glycerin, transferinto a glass-stoppered ‰ask, add 50 mL of water, mix, addexactly 50 mL of sodium periodate TS, shake, and allow tostand in a dark place at a room temperature for about 30minutes. Add 10 mL of a mixture of water and ethyleneglycol (1:1), allow to stand for about 20 minutes, add 100mL of water, and titrate with 0.1 mol/L sodium hydroxideVS (indicator: 2 drops of phenolphthalein TS). Perform ablank determination, and make the necessary correction.


Concentrated Glycerin濃グリセリン


Concentrated Glycerin contains not less than 98.0zand not more than 101.0z of glycerin (C3H8O3), cal-culated of the anhydrous basis.


Description Concentrated Glycerin is a clear, colorless andviscous liquid. It has a sweet taste.

It is miscible with water and with ethanol (99.5).It is hygroscopic.


Identiˆcation Determine the infrared absorption spectrumof Concentrated Glycerin as directed in the liquid ˆlmmethod under the Infrared Spectrophotometry, and com-pare the spectrum with the Reference Spectrum: both spec-tra exhibit similar intensities of absorption at the same wavenumbers.

Add the following next to Purity:



Add the following next to Residue on ignition:

Assay Weigh accurately about 0.2 g of Concentrated Glyc-erin, transfer into a glass-stoppered ‰ask, add 50 mL ofwater, mix, add exactly 50 mL of sodium periodate TS,shake, and allow to stand in a dark place at a room tempera-ture for about 30 minutes. Add 10 mL of a mixture of waterand ethylene glycol (1:1), allow to stand for about 20minutes, add 100 mL of water, and titrate with 0.1 mol/Lsodium hydroxide VS (indicator: 2 drops of phenolphthaleinTS). Perform a blank determination, and make the necessa-ry correction.


Add the following:

Gramicidinグラミシジン

[1405-97-6]

Gramicidin is a mixture of peptide substances hav-ing antibacterial activity produced by the growth ofBacillus brevis Dubos.

It contains not less than 900 mg (potency) per mg,calculated on the dried basis. The potency ofGramicidin is expressed as mass (potency) ofgramicidin.

Description Gramicidin occurs as a white to light yellowishwhite crystalline powder.

It is freely soluble in methanol, soluble in ethanol (99.5),and practically insoluble in water.

Identiˆcation (1) To 10 mg of Gramicidin add 2 mL of6 mol/L hydrochloric acid TS, and heat in a water bath for30 minutes with occasional stirring. After cooling, neutralizewith 6 mol/L sodium hydroxide TS, add 1 mL of ninhydrinTS and 0.5 mL of pyridine, and heat for 2 minutes: a blue-purple to red-purple color develops.

(2) Determine the absorption spectrum of a solution ofGramicidin in ethanol (95) (1 in 20,000), as directed underthe Ultraviolet-visible Spectrophotometry, and compare thespectrum with the Reference Spectrum or the spectrum of asolution of Gramicidin Reference Standard prepared in thesame manner as the sample solution: both spectra exhibitsimilar intensities of absorption at the same wavelengths.

Loss on drying Not more than 3.0z (0.1 g, in vacuum,609C, 3 hours).


Assay Perform the test according to the Turbidimetricmethod as directed under the Microbial Assay for Antibiot-ics according to the following conditions.

(1) Test organism—Enterococcus hirae ATCC 10541(2) Agar medium for transferring test organism

Glucose 10.0 gCasein peptone 5.0 gYeast extract 20.0 gPotassium dihydrogen phosphate 2.0 gPolysorbate 80 0.1 gAgar 15.0 gWater 1000 mL


(3) Liquid medium for suspending test organism—Usethe culture medium (2).

(4) Preparation of the test organism suspension—Pun-cture the test organism in the medium, prepared by dispens-ing 10 mL of the agar medium for transferring test organismin a test tube about 16 mm in inside diameter, incubate at36.5 to 37.59C for 20 to 24 hours. After sub-culturing atleast three times, keep between 1 to 59C. Transfer the organ-ism so obtained in 10 mL of the liquid medium for suspend-ing test organism, incubate at 36.5 to 37.59C for 20 to 24hours, and use this medium as the test organism stock sus-pension. Before use, add the test organism stock suspensionto the liquid medium for suspending test organism so thatthe transmittance at 580 nm is 50 to 60z. Mix one volume ofthis suspension and 200 volume of the liquid medium forsuspending test organism, and use this as the test organismsuspension.

(5) Standard solution—Weigh accurately an amount ofGramicidin Reference Standard, previously dried underreduced pressure not exceeding 0.67 kPa at 609C for 3hours, equivalent to about 10 mg (potency), dissolve inethanol (99.5) to make exactly 100 mL, and use this solutionas the standard stock solution. Keep the standard stock solu-tion at not exceeding 59C and use within 7 days. Take ex-actly a suitable amount of the standard stock solution beforeuse, add the following diluting solution to make a solutionso that each mL contains 0.02 mg (potency), and use thissolution as the standard solution.

Diluting solution: To 390 mL of propylene glycol add 210mL of a mixture of ethanol (99.5) and acetone (9:1) andSterile Puriˆed Water to make 1000 mL.

(6) Sample solution—Weigh accurately an amount ofGramicidin, equivalent to about 10 mg (potency), and dis-solve in ethanol (99.5) to make exactly 100 mL. Take exactlya suitable amount of this solution, add the diluting solutionobtained in (5) to make a solution so that each mL contains0.02 mg (potency), and use this solution as the sample solu-tion.

(7) Procedure—Transfer 0.155 mL, 0.125 mL, 0.100mL, 0.080 mL and 0.065 mL each of the standard solution,0.100 mL of the sample solution and 0.100 mL of the dilut-ing solution, separately, in test tubes about 14 mm in insidediameter and about 15 cm in length, and make three sets foreach. To each of the test tube add 10 mL of the test organismsuspension, stopper the tube, incubate in a water bath at36.5 to 37.59C for 180 to 270 minutes, add 0.5 mL of a solu-tion of formaldehyde (1 in 3), and determine their transmit-tances at 580 nm.



Griseofulvinグリセオフルビン


Griseofulvin contains not less than 960 mg (potency)per mg, calculated on the dried basis. The potencyof Griseofulvin is expressed as mass (potency) ofgriseofulvin (C17H17ClO6).

Description Griseofulvin occurs as white, crystals or crys-talline powder.

It is soluble in N,N-dimethylformamide, sparingly solublein acetone, slightly soluble in methanol and in ethanol (95),and practically insoluble in water.

Identiˆcation (1) Determine the absorption spectrum ofa solution of Griseofulvin in ethanol (95) (1 in 100,000) asdirected under the Ultraviolet-visible Spectrophotometry,and compare the spectrum with the Reference Spectrum orthe spectrum of a solution of Griseofulvin Reference Stan-dard prepared in the same manner as the sample solution:both spectra exhibit similar intensities of absorption at thesame wavelengths.

(2) Determine the infrared absorption spectrum ofGriseofulvin as directed in the potassium bromide diskmethod under the Infrared Spectrophotometry, and com-pare the spectrum with the Reference Spectrum or thespectrum of Griseofulvin Reference Standard: both spectraexhibit similar intensities of absorption at the same wavenumbers.


the dried basis, N,N-dimethylformamide, 25 mL, 100 mm).


Purity (1) Acid—Dissolve 0.25 g of Griseofulvin in20 mL of neutralized ethanol, and add 2 drops ofphenolphthalein TS and 1.0 mL of 0.02 mol/L sodiumhydroxide VS: the color of the solution is red.

(2) Heavy metals—Proceed with 1.0 g of Griseofulvinaccording to Method 2, and perform the test. Prepare thecontrol solution with 2.5 mL of Standard Lead Solution (notmore than 25 ppm).

(3) Arsenic—Prepare the test solution with 1.0 g ofGriseofulvin according to Method 3, and perform the test(not more than 2 ppm).

(4) Related substances—To 0.10 g of Griseofulvin addexactly 1 mL of the internal standard solution and acetone tomake 10 mL, and use this solution as the sample solution.Separately, to 5.0 mg of Griseofulvin Reference Standardadd exactly 1 mL of the internal standard solution and ace-tone to make 10 mL, and use this solution as the standardsolution. Perform the test with 2 mL each of the sample solu-

tion and the standard solution as directed under the GasChromatography according to the following conditions,determine each peak area by the automatic integrationmethod, and calculate the ratio, Q1, of the peak area ofdechlorogriseofulvin, having the relative retention time ofabout 0.6 with respect to griseofulvin, to that of the internalstandard obtained from the sample solution, the ratio, Q2,of the peak area of dehydrogriseofulvin, having the relativeretention time of about 1.2 with respect to griseofulvin, tothat of the internal standard obtained from the sample solu-tion and the ratio, QS, of the peak area of griseofulvin tothat of the internal standard obtained from the standardsolution: Q1/QS is not more than 0.6, and Q2/QS is not morethan 0.15.Internal standard solution—A solution of 9,10-diphenylanthracene in acetone (1 in 500).Operating conditions—

Detector: An hydrogen ‰ame-ionization detector.Column: A glass column 4 mm in inside diameter and 1 m

in length, packed with siliceous earth for gas chro-matography coated with 25z phenyl-25z cyanopropyl-methylsilicone polymer for gas chromatography in the ratioof 1z (150 – 180 mm in particle diameter).


Temperature of injection port: A constant temperature ofabout 2709C.

Temperature of detector: A constant temperature ofabout 3009C.


of griseofulvin is about 10 minutes.System suitability—

Test for required detectability: Measure exactly 1 mL ofthe standard solution, and add the internal standard solutiondiluted with acetone (1 in 10) to make exactly 10 mL. Con-ˆrm that the ratio of the peak area of griseofulvin to that ofthe internal standard obtained from 2 mL of this solution isequivalent to 7 to 13z of that obtained from 2 mL of thestandard solution.

System performance: When the procedure is run with 2 mLof the standard solution under the above operating condi-tions, the internal standard and griseofulvin are eluted inthis order with the resolution between these peaks being notless than 5.

System repeatability: When the test is repeated 6 timeswith 2 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratiosof the peak area of griseofulvin to that of the internal stan-dard is not more than 5.0z.

(5) Petroleum ether soluble substances—To 1.0 g ofGriseofulvin add 20 mL of petroleum ether, shake, and boilfor 10 minutes under a re‰ux condenser. After cooling, ˆlterthrough a dried ˆlter paper, wash the ˆlter paper with two15-mL portions of petroleum ether, combine the washings tothe ˆltrate, evaporate the petroleum ether on a water bath,and dry the residue at 1059C for 1 hour: the amount of theresidue is not more than 0.2z.




Assay Weigh accurately an amount of Griseofulvin andGriseofulvin Reference Standard, equivalent to about 50 mg(potency), dissolve each in 50 mL of N,N-dimethylform-amide, add exactly 20 mL of the internal standard solutionand water to make 250 mL, and use these solutions as thesample solution and the standard solution. Perform the testwith exactly 10 mL each of the sample solution and the stan-dard solution as directed under the Liquid Chromatographyaccording to the following conditions, and determine theratios, QT and QS, of the peak area of griseofulvin to that ofthe internal standard.

Amount [mg (potency)] of C17H17ClO6＝WS×QT

QS× 1000

WS: Amount [mg (potency)] of Griseofulvin ReferenceStandard

Internal standard solution—A solution of butyl parahydrox-ybenzoate in acetonitrile (1 in 400).Operating conditions—




Mobile phase: A mixture of water and acetonitrile (3:2).Flow rate: Adjust the ‰ow rate so that the retention time

of griseofulvin is about 6 minutes.System suitability—

System performance: When the procedure is run with10 mL of the standard solution under the above operatingconditions, griseofulvin and the internal standard are elutedin this order with the resolution between these peaks beingnot less than 4.

System repeatability: When the test is repeated 6 timeswith 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratiosof the peak area of griseofulvin to that of the internal stan-dard is not more than 1.0z.


Haloxazolamハロキサゾラム


Purity(5) Related substances—Dissolve 0.10 g of Haloxazolam

in 100 mL of acetonitrile, and use this solution as the sample

solution. Pipet 1 mL of the sample solution, add acetonitrileto make exactly 100 mL, and use this solution as the stan-dard solution. Perform the test with exactly 10 mL each ofthe sample solution and the standard solution as directed un-der the Liquid Chromatography according to the followingconditions. Determine each peak area of both solutions bythe automatic integration method: the total area of all peaksother than the area of the haloxazolam from the samplesolution is not larger than the peak area of the haloxazolamfrom the standard solution.Operating conditions—




Mobile phase: Dissolve 6.2 g of boric acid and 7.5 g ofpotassium chloride in 900 mL of water, adjust the pH withtriethylamine to 8.5, and add water to make 1000 mL. To300 mL of this solution add 200 mL of acetonitrile.

Flow rate: Adjust the ‰ow rate so that the retention timeof haloxazolam is about 10 minutes.

Time span of measurement: About 3 times as long as theretention time of haloxazolam after the solvent peak.System suitability—

Test for required detection: To exactly 5 mL of the stan-dard solution add acetonitrile to make exactly 50 mL. Con-ˆrm that the peak area of haloxazolam obtained from 10 mLof this solution is equivalent to 8 to 12z of that of halox-azolam obtained from 10 mL of the standard solution.

System performance: Dissolve 10 mg each of Haloxazol-am and cloxazolam in 200 mL of acetonitrile. When theprocedure is run with 10 mL of this solution under the aboveoperating conditions, haloxazolam and cloxazolam are elut-ed in this order with the resolution between these peaksbeing not less than 1.5.

System repeatability: When the test is repeated 6 timeswith 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peakarea of haloxazolam is not more than 1.0z.

Homochlorcyclizine Hydrochloride塩酸ホモクロルシクリジン


Description Homochlorcyclizine Hydrochloride occurs aswhite to pale brown, crystals or powder.

It is very soluble in water, freely soluble in acetic acid(100), slightly soluble in ethanol (99.5), and very slightlysoluble in acetonitrile and in acetic anhydride.

It dissolves in 0.1 mol/L hydrochloric acid TS.It is hygroscopic.


It is colored slightly by light.A solution of Homochlorcyclizine Hydrochloride (1 in 10)

shows no optical rotation.Melting point: about 2279C (with decomposition).


Identiˆcation (1) Determine the absorption spectrumof a solution of Homochlorcyclizine Hydrochloride in0.1 mol/L hydrochloric acid TS (1 in 100,000) as directedunder the Ultraviolet-visible Spectrophotometry, and com-pare the spectrum with the Reference Spectrum: both spec-tra exhibit similar intensities of absorption at the same wave-lengths.

(2) Determine the infrared absorption spectrum ofHomochlorcyclizine Hydrochloride, previously dried, asdirected in the potassium chloride disk method under the In-frared Spectrophotometry, and compare the spectrum withthe Reference Spectrum: both spectra exhibit similar intensi-ties of absorption at the same wave numbers.

(3) A solution of Homochlorcyclizine Hydrochloride (1in 100) responds to the Qualitative Tests for chloride.


Purity (1) Heavy metals—Proceed with 1.0 g of Homo-chlorcyclizine Hydrochloride according to Method 2, andperform the test. Prepare the control solution with 2.0 mLof Standard Lead Solution (not more than 20 ppm).

(2) Related substances—Dissolve 0.10 g of Homochlor-cyclizine Hydrochloride in 100 mL of the mobile phase, anduse this solution as the sample solution. Measure exactly1 mL of this solution, add the mobile phase to make exactly100 mL, and use this solution as the standard solution.Perform the test with exactly 10 mL each of the sample solu-tion and the standard solution as directed under the LiquidChromatography according to the following conditions, anddetermine the peak areas by the automatic integrationmethod: the areas of the peaks other than homochlorcycli-zine obtained from the sample solution are not more than1/2 times the peak area of homochlorcyclizine from thestandard solution, and the total area of the peaks other thanhomochlorcyclizine from the sample solution is not morethan the peak area of homochlorcyclizine from the standardsolution.Operating conditions—




Mobile phase: A mixture of water, acetonitrile and per-chloric acid (134:66:1).

Flow rate: Adjust the ‰ow rate so that the retention timeof homochlorcyclizine is about 10 minutes.

Time span of measurement: About 2 times as long as the

retention time of homochlorcyclizine.System suitability—

Test for required detection: To exactly 5 mL of the stan-dard solution add the mobile phase to make exactly 50 mL.Conˆrm that the peak area of homochlorcyclizine obtainedfrom 10 mL of this solution is equivalent to 7 to 13z of thatof homochlorcyclizine obtained from 10 mL of the standardsolution.

System performance: Dissolve 5 mg each of Homochlor-cyclizine Hydrochloride and methyl parahydroxybenzoicacid in 100 mL of the mobile phase. When the procedure isrun with 10 mL of this solution under the above operatingconditions, methyl parahydroxybenzoic acid and homo-chlorcyclizine are eluted in this order with the resolutionbetween these peaks being not less than 5.

System repeatability: When the test is repeated 6 timeswith 10mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peakarea of homochlorcyclizine is not more than 1.0z.

Hydrochlorothiazideヒドロクロロチアジド


Assay Weigh accurately about 30 mg each of Hydro-chlorothiazide and Hydrochlorothiazide Reference Stan-dard, previously dried, and dissolve in 150 mL of the mobilephase, add exactly 10 mL each of the internal standard solu-tion, then add the mobile phase to make 200 mL, and usethese solutions as the sample solution and the standard solu-tion, respectively. Perform the test with 20 mL each of thesample solution and the standard solution as directed underthe Liquid Chromatography according to the following con-ditions, and calculate the ratios, QT and QS, of the peak areaof hydrochlorothiazide to that of the internal standard.

Amount (mg) of C7H8ClN3O4S2＝WS×QT

QS

WS: Amount (mg) of Hydrochlorothiazide ReferenceStandard

Internal standard solution—A solution of 4-aminoacetophe-none in acetonitrile (9 in 2000).Operating conditions—




Mobile phase: A mixture of 0.1 mol/L sodium dihydro-gen phosphate TS, pH 3.0 and acetonitrile (9:1).

Flow rate: Adjust the ‰ow rate so that the retention timeof hydrochlorothiazide is about 10 minutes.



20 mL of the standard solution under the above operatingconditions, hydrochlorothiazide and the internal standardare eluted in this order with the resolution between thesepeaks being not less than 4.

System repeatability: When the test is repeated 6 timeswith 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratiosof the peak area of hydrochlorothiazide to that of the inter-nal standard is not more than 1.0z.

Hydrocortisone Acetate酢酸ヒドロコルチゾン


Assay Dissolve about 20 mg each of HydrocortisoneAcetate and Hydrocortisone Acetate Reference Standard,previously dried and accurately weighed, in methanol, addexactly 10 mL each of the internal standard solution, thenadd methanol to make 100 mL, and use these solutions asthe sample solution and the standard solution. Perform thetest with 20 mL each of the sample solution and the standardsolution as directed under the Liquid Chromatography ac-cording to the following conditions, and calculate the ratios,QT and QS, of the peak area of hydrocortisone acetate tothat of the internal standard, respectively.


QS

WS: Amount (mg) of Hydrocortisone Acetate ReferenceStandard

Internal standard solution—A solution of benzyl para-hydroxybenzoate in methanol (1 in 1000).Operating conditions—





of hydrocortisone acetate is about 8 minutes.System suitability—

System performance: When the procedure is run with20 mL of the standard solution under the above operatingconditions, hydrocortisone acetate and the internal standardare eluted in this order with the resolution between thesepeaks being not less than 4.

System repeatability: When the test is repeated 6 timeswith 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratios

of the peak area of hydrocortisone acetate to that of theinternal standard is not more than 1.0z.

Hydrocortisone Sodium Phosphateリン酸ヒドロコルチゾンナトリウム

Cange the Purity (6) to read:

Purity(6) Free hydrocortisone—Dissolve 25 mg of Hydrocorti-

sone Sodium Phosphate in the mobile phase to make exactly20 mL, and use this solution as the sample solution.Separately, weigh 25 mg of Hydrocortisone Reference Stan-dard, previously dried at 1059C for 3 hours, and dissolve inthe mobile phase to make exactly 100 mL. Pipet 10 mL ofthis solution, add the mobile phase to make exactly 200 mL,and use this solution as the standard solution. Perform thetest with 20 mL each of the sample solution and the standardsolution as directed under the Liquid Chromatography ac-cording to the following conditions. Determine the peakareas, AT and AS, of hydrocortisone from each solution: AT

is not larger than AS.Operating conditions—



System repeatability: When the test is repeated 6 timeswith 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peakarea of hydrocortisone is not more than 1.0z.


Assay Weigh accurately about 20 mg each of Hydrocorti-sone Sodium Phosphate and Hydrocortisone Sodium Phos-phate Reference Standard (determine its loss on dryingbefore using), dissolve each in 50 mL of the mobile phase,add exactly 10 mL of the internal standard solution, thenadd the mobile phase to make 200 mL, and use these solu-tions as the sample solution and the standard solution,respectively. Perform the test with 20 mL each of the samplesolution and the standard solution as directed under the Liq-uid Chromatography according to the following conditions,and calculate the ratios, QT and QS, of the peak area ofhydrocortisone phosphate to that of the internal standard,respectively.

Amount (mg) of C21H29Na2O8P＝ WS×QT

QS

WS: Amount (mg) of Hydrocortisone Sodium PhosphateReference Standard, calculated on the dried basis

Internal standard solution—A solution of isopropyl para-hydroxybenzoate in the mobile phase (3 in 5000).






Mobile phase: A mixture of 0.05 mol/L sodium dihydro-gen phosphate TS, pH 2.6 and methanol (1:1).

Flow rate: Adjust the ‰ow rate so that the retention timeof hydrocortisone phosphate is about 10 minutes.System suitability—

System performance: When the procedure is run with20 mL of the standard solution under the above operatingconditions, hydrocortisone phosphate and isopropyl para-hydroxybenzoate are eluted in this order with the resolutionbetween these peaks being not less than 8.

System repeatability: When the test is repeated 6 timeswith 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratiosof the peak area of hydrocortisone phosphate to that of theinternal standard is not more than 1.0z.

Idarubicin Hydrochloride塩酸イダルビシン



Idoxuridine Ophthalmic Solutionイドクスウリジン点眼液


Purity 5-Iodouracil and 2?-deoxyuridine—To a volume ofIdoxuridine Ophthalmic Solution, equivalent to 4.0 mg ofIdoxuridine according to the labeled amount, add water tomake exactly 5 mL, and use this solution as the sample solu-tion. Separately, dissolve 12.0 mg of 5-iodouracil for liquidchromatography and 4.0 mg of 2?-deoxyuridine for liquidchromatography in water to make exactly 200 mL. Measureexactly 5 mL of this solution, add water to make exactly25 mL, and use this solution as the standard solution.Perform the test with exactly 10 mL each of the sample solu-tion and the standard solution as directed under the LiquidChromatography according to the following conditions, anddetermine the peak areas of 5-iodouracil and 2?-deoxyuri-dine: the peak areas of 5-iodouracil and 2?-deoxyuridine ofthe sample solution are not more than the peak areas of 5-iodouracil and 2?-deoxyuridine of the standard solution.Operating conditions—






of 2?-deoxyuridine is about 6 minutes.System suitability—System performance: When the procedure is run with 10 mLof the standard solution under the above operating condi-tions, 2?-deoxyuridine and 5-iodouracil are eluted in thisorder with the resolution between these peaks being not lessthan 2.0.

System repeatability: When the test is repeated 6 timeswith 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peakarea of 2?-deoxyuridine is not more than 1.0z.


Assay Measure exactly a volume of IdoxuridineOphthalmic Solution, equivalent to 3 mg of idoxuridine(C9H11IN2O5) according to the labeled amount, add exactly2 mL of the internal standard solution, then add water tomake 10 mL, and use this solution as the sample solution.Separately weigh accurately about 10 mg of IdoxuridineReference Standard, previously dried at 609C for 3 hours,dissolve in water to make exactly 10 mL. Measure exactly3 mL of this solution, add exactly 2 mL of the internal stan-dard solution, then add water to make 10 mL, and use thissolution as the standard solution. Perform the test with10 mL each of the sample solution and the standard solutionas directed under the Liquid Chromatography according tothe following conditions, and calculate the ratios, QT andQS, of the peak area of idoxuridine to that of the internalstandard, respectively.

Amount (mg) of idoxuridine (C9H11IN2O5)

＝WS×QT

QS×

310

WS: Amount (mg) of Idoxuridine Reference Standard

Internal standard solution—A solution of sulfathiazole inthe mobile phase (1 in 4000).Operating conditions—





of idoxuridine is about 9 minutes.



10 mL of the standard solution under the above operatingconditions, idoxuridine and the internal standard are elutedin this order with the resolution between these peaks beingnot less than 2.0.

System repeatability: When the test is repeated 6 timeswith 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratiosof the peak area of idoxuridine to that of the internal stan-dard is not more than 1.0z.

Imipenemイミペネム


Imipenem contains not less than 924 mg (potency)per mg, calculated on the anhydrous basis. The poten-cy of Imipenem is expressed as mass (potency) of im-ipenem (C12H17N3O4S: 299.35).

Description Imipenem occurs as white to light yellow crys-talline powder.

It is sparingly soluble in water, and practically insoluble inethanol (99.5).

Identiˆcation (1) Determine the absorption spectrum ofa solution of Imipenem in 0.1 mol/L 3-(N-mor-pholino)propanesulfonic acid buŠer solution, pH 7.0 (1 in50,000) as directed under the Ultraviolet-visible Spec-trophotometry, and compare the spectrum with the Refer-ence Spectrum or the spectrum of a solution of ImipenemReference Standard prepared in the same manner as thesample solution: both spectra exhibit similar intensities ofabsorption at the same wavelengths.

(2) Determine the infrared absorption spectrum of Im-ipenem as directed in the potassium bromide disk methodunder the Infrared Spectrophotometry, and compare thespectrum with the Reference Spectrum or the spectrum ofImipenem Reference Standard: both spectra exhibit similarintensities of absorption at the same wave numbers.

Optical rotation [a]20D :＋84 –＋899(50 mg calculated on the

anhydrous basis, 0.1 mol/L 3-(N-morpholino)propanesul-fonic acid buŠer solution, pH 7.0, 10 mL, 100 mm).

pH The pH of a solution obtained by dissolving 1.0 g ofImipenem in 200 mL of water is between 4.5 and 7.0.

Purity (1) Heavy metals—Proceed with 1.0 g of Imipen-em according to Method 2, and perform the test. Prepare thecontrol solution with 2.0 mL of Standard Lead Solution (notmore than 20 ppm).

(2) Arsenic—Put 2.0 g of Imipenem in a crucible, add 5mL of nitric acid and 1 mL of sulfuric acid, and heat careful-ly until white fumes evolve. After cooling, add 2 mL of nitric

acid, heat, and repeat this procedure once more. Then add 2mL of hydrogen peroxide (30), heat, and repeat this proce-dure several times until the color of the solution changes tocolorless to pale yellow. After cooling, heat again until whitefumes evolve. After cooling, add water to make 5 mL, andperform the test with this solution as the test solution (notmore than 1 ppm).

(3) Related substances—Dissolve 50 mg of Imipenem in50 mL of 0.1 mol/L 3-(N-morpholino)propanesulfonic acidbuŠer solution, pH 7.0, and use this solution as the samplesolution. Pipet 1 mL of the sample solution, add 0.1 mol/L3-(N-morpholino)propanesulfonic acid buŠer solution, pH7.0 to make exactly 100 mL, and use this solution as thestandard solution. Perform the test with exactly 10 mL eachof the sample solution and the standard solution as directedunder the Liquid Chromatography according to the follow-ing conditions, and determine each peak area by the auto-matic integration method: the peak area of thienamycin,having the relative retention time of about 0.8 with respectto imipenem, obtained from the sample solution is not morethan the peak area of imipenem from the standard solution,the area of the peak other than imipenem and thienamycinfrom the sample solution is not more than 1/3 times the peakarea of imipenem from the standard solution, and the totalarea of the peaks other than imipenem and thienamycinfrom the sample solution is not more than the peak area ofimipenem from the standard solution.Operating conditions—


Time span of measurement: About 2 times as long as theretention time of imipenem.System suitability—

Test for required detectability: Measure exactly 5 mL ofthe standard solution, add 0.1 mol/L 3-(N-morpholino)-propanesulfonic acid buŠer solution, pH 7.0 to make exactly50 mL. Conˆrm that the peak area of imipenem from 10 mLof this solution is equivalent to 7 to 13z of that from thestandard solution.


System repeatability: When the test is repeated 6 timeswith 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peakarea of imipenem is not more than 2.0z.

Water Not less than 5.0z and not more than 8.0z (20mg, coulometric titration, water evaporation temperature:1409C).


Assay Weigh accurately an amount of Imipenem and Im-ipenem Reference Standard, equivalent to about 50 mg(potency), dissolve each in 0.1 mol/L 3-(N-morpholino)-propanesulfonic acid buŠer solution, pH 7.0 to make exactly50 mL, and use these solutions as the sample solution andthe standard solution. Perform the test with exactly 10 mL


each of the sample solution and the standard solution,within 30 minutes after preparation of these solutions, asdirected under the Liquid Chromatography according to thefollowing conditions, and determine the peak areas, AT andAS, of imipenem of these solutions.


＝WS×AT

AS× 1000

WS: Amount [mg (potency)] of Imipenem ReferenceStandard

Operating conditions—Detector: An ultraviolet absorption photometer

(wavelength: 280 nm).Column: A stainless steel column 3.9 mm in inside di-



Mobile phase: A mixture of 0.1 mol/L 3-(N-mor-pholino)propanesulfonic acid buŠer solution, pH 7.0 andacetonitrile (100:1).

Flow rate: Adjust the ‰ow rate so that the retention timeof imipenem is about 6 minutes.System suitability—

System performance: Dissolve 50 mg of Imipenem and 75mg of resorcinol in 50 mL of 0.1 mol/L 3-(N-mor-pholino)propanesulfonic acid buŠer solution, pH 7.0. Whenthe procedure is run with 10 mL of this solution under theabove operating conditions, imipenem and resorcinol areeluted in this order with the resolution between these peaksbeing not less than 4.

System repeatability: When the test is repeated 5 timeswith 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peakarea of imipenem is not more than 0.80z.


Indometacin Suppositoriesインドメタシン坐剤


Assay Weigh accurately not less than 20 Indometacin Sup-positories, cut into small pieces carefully, and mix well.Weigh accurately a portion of the mass, equivalent to about50 mg of indometacin (C19H16ClNO4), add 40 mL of tetra-hydrofuran, warm at 409C, dissolve by shaking, cool, andadd tetrahydrofuran to make exactly 50 mL. Filter the solu-tion, discard the ˆrst 10 mL of the ˆltrate, pipet the subse-quent 5 mL of the ˆltrate, add exactly 3 mL of the internalstandard solution, and add the mobile phase to make100 mL. Allow the solution to stand for 30 minutes, ˆlterthrough a membrane ˆlter (0.5 mm pore size), discard theˆrst 10 mL of the ˆltrate, and use the subsequent ˆltrate as

the sample solution. Separately, weigh accurately about50 mg of Indometacin Reference Standard, previously driedat 1059C for 4 hours, and dissolve in tetrahydrofuran tomake exactly 50 mL. Pipet 5 mL of the solution, proceed inthe same manner as the sample solution, and use as the stan-dard solution. Perform the test with 20 mL each of the sam-ple solution and the standard solution as directed under theLiquid Chromatography according to the following condi-tions, and calculate the ratios, QT and QS, of the peak areaof indometacin to that of the internal standard, respectively.

Amount (mg) of indometacin (C19H16ClNO4)

＝WS×QT

QS

WS: Amount (mg) of Indometacin Reference Standard


Detector: An ultraviolet absorption photometer(wavelength: 254 nm).



Mobile phase: A mixture of methanol and diluted phos-phoric acid (1 in 1000) (7:3).

Flow rate: Adjust the ‰ow rate so that the retention timeof indometacin is about 8 minutes.System suitability—

System performance: Dissolve 50 mg of 4-chlorobenzoicacid, 30 mg of butyl parahydroxybenzoate and 50 mg of in-dometacin in 50 mL of methanol. To 5 mL of this solutionadd the mobile phase to make 100 mL. When the procedureis run with 20 mL of this solution under the above operatingconditions, 4-chlorobenzoic acid, butyl parahydroxybenzo-ate and indometacin are eluted in this order with the resolu-tion between the peaks of 4-chlorobenzoic acid and butylparahydroxybenzoate being not less than 2.0 and betweenthe peaks of parahydroxybenzoate and indometacin beingnot less than 5.

System repeatability: When the test is repeated 6 timeswith 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratiosof the peak area of indometacin to that of the internal stan-dard is not more than 1.0z.



Iopanoic Acidイオパノ酸

Iopanoic Acid Tabletsイオパノ酸錠

Isoniazid Injectionイソニアジド注射液


Description Isoniazid Injection occurs as a clear, colorlessliquid.

pH: 6.5 – 7.5.


Identiˆcation To a volume of Isoniazid Injection, equiva-lent to 20 mg of Isoniazid according to the labeled amount,and add water to make 200 mL. To 5 mL of the solution add1 mL of 0.1 mol/L hydrochloric acid TS and water to make50 mL. Determine the absorption spectrum of this solutionas directed under the Ultraviolet-visible Spectrophotometry:it exhibits a maximum between 264 nm and 268 nm.


Assay To an exactly measured volume of Isoniazid Injec-tion, equivalent to about 50 mg of isoniazid (C6H7N3O), addwater to make exactly 100 mL. Pipet 5 mL of the solution,add exactly 5 mL of the internal standard and the mobilephase to make 50 mL, and use this solution as the sample so-lution. Separately, weigh accurately about 50 mg of isonia-zid for assay, previously dried at 1059C for 2 hours, and dis-solve in water to make exactly 100 mL. Pipet 5 mL of thissolution, add exactly 5 mL of the internal standard and themobile phase to make 50 mL, and use this solution as thestandard solution. Perform the test with 5 mL each of thesample solution and the standard solution as directed underthe Liquid Chromatography according to the following con-ditions, and determine the ratios, QT and QS, of the peakarea of isoniazid to that of the internal standard.

Amount (mg) of isoniazid (C6H7N3O)＝WS×QT

QS

WS: Amount (mg) of isoniazid for assay

Internal standard solution—A solution of propyl para-hydroxybenzoate (1 in 4000).Operating conditions—


Column: A stainless steel column 4.6 mm in inside di-



Mobile phase: Dissolve 6.80 g of potassium dihydrogenphosphate in water to make 1000 mL. Separately, to 5.76 gof phosphoric acid add water to make 1000 mL. Mix thesesolutions to make a solution having pH 2.5. To 500 mL ofthis solution add 500 mL of methanol, and add 2.86 g of so-dium tridecanesulfonate to dissolve.

Flow rate: Adjust the ‰ow rate so that the retention timeof isoniazid is about 5 minutes.System suitability—

System performance: When the procedure is run with 5 mLof the standard solution under the above operating condi-tions, isoniazid and the internal standard are eluted in thisorder with the resolution between these peaks being not lessthan 10.

System repeatability: When the test is repeated 6 timeswith 5 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratiosof the peak area of isoniazid to that of the internal standardis not more than 1.3z.

Isoniazid Tabletsイソニアジド錠


Assay Weigh accurately and powder not less than 20Isoniazid Tablets. Weigh accurately a quantity of the pow-der, equivalent to about 0.10 g of isoniazid (C6H7N3O), add150 mL of water, shake for 30 minutes, then add water tomake exactly 200 mL, and ˆlter. Discard the ˆrst 10 mL ofthe ˆltrate, pipet 5 mL of the subsequent ˆltrate, add themobile phase to make exactly 50 mL, and use this solution asthe sample solution. Separately, weigh accurately about 50mg of isoniazid for assay, previously dried at 1059C for 2hours, dissolve in water to make exactly 100 mL. Pipet 5 mLof this solution, add the mobile phase to make exactly 50mL, and use this solution as the standard solution. Performthe test with 10 mL each of the sample solution and the stan-dard solution as directed under the Liquid Chromatographyaccording to the following conditions. Determine the peakareas, AT and AS, of isoniazid of the sample solution and thestandard solution.

Amount (mg) of isoniazid (C6H7N3O)＝WS×AT

AS× 2

WS: Amount (mg) of isoniazid for assay



ameter and 25 cm in length, packed with octadecylsilanized




Mobile phase: Dissolve 6.80 g of potassium dihydrogenphosphate in water to make 1000 mL. Separately, to 5.76 gof phosphoric acid add water to make 1000 mL. Mix thesesolutions to adjust the pH to 2.5. To 400 mL of this solutionadd 600 mL of methanol, and dissolve 2.86 g of sodiumtridecanesulfonate in this.

Flow rate: Adjust the ‰ow rate so that the retention timeof isoniazid is about 5 minutes.System suitability—

System performance: Dissolve 5 mg of Isoniazid and 5 mgof isonicotinic acid in 100 mL of the mobile phase. When theprocedure is run with 10 mL of this solution under the aboveoperating conditions, isonicotinic acid and isoniazid areeluted in this order with the resolution between these peaksbeing not less than 1.5.

System repeatability: When the test is repeated 6 timeswith 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peakarea of isoniazid is not more than 1.0z.

Josamycinジョサマイシン


Josamycin contains not less than 900 mg (potency)per mg, calculated on the dried basis. The potency ofJosamycin is expressed as mass (potency) of josamycin(C42H69NO15).

Description Josamycin occurs as a white to yellowish whitepowder.

It is very soluble in methanol and in ethanol (99.5), andvery slightly soluble in water.

Identiˆcation (1) Determine the absorption spectrum ofa solution of Josamycin in methanol (1 in 100,000) as direct-ed under the Ultraviolet-visible Spectrophotometry, andcompare the spectrum with the Reference Spectrum or thespectrum of a solution of Josamycin Reference Standardprepared in the same manner as the sample solution: bothspectra exhibit similar intensities of absorption at the samewavelengths.

(2) Dissolve 5 mg each of Josamycin and JosamycinReference Standard in 1 mL of methanol, add dilutedmethanol (1 in 2) to make 100 mL, and use these solutions asthe sample solution and the standard solution, respectively.Perform the test with 10 mL each of the sample solution andthe standard solution as directed under the Liquid Chro-matography according to the following conditions: theretention time of the main peak obtained from the samplesolution is the same with that of the peak of josamycin from

the standard solution.Operating conditions—

Detector, column, column temperature, mobile phase,and ‰ow rate: Proceed as directed in the operating condi-tions in the Purity (2).

Purity (1) Heavy metals—Proceed with 1.0 g of Josamy-cin according to Method 2, and perform the test. Prepare thecontrol solution with 3.0 mL of Standard Lead Solution (notmore than 30 ppm).

(2) Related substances—Dissolve 50 mg of Josamycin in5 mL of methanol, add diluted methanol (1 in 2) to make 50mL, and use this solution as the sample solution. Performthe test with 10 mL of the sample solution as directed underthe Liquid Chromatography according to the following con-ditions. Determine each peak area by the automatic integra-tion method, and calculate the amounts of josamycin andthe related substances by the area percentage method: theamount of any peak other than josamycin is not more than6z, and the total of these peaks is not more than 20z.Operating conditions—




Mobile phase: Dissolve 119 g of sodium perchlorate inwater to make 1000 mL, and adjust the pH to 2.5 with 1mol/L hydrochloric acid TS. To 600 mL of this solution add400 mL of acetonitrile.

Flow rate: Adjust the ‰ow rate so that the retention timeof josamycin is about 10 minutes.

Time span of measurement: About 4 times as long as theretention time of josamycin after the solvent peak.System suitability—

Test for required detectability: Pipet 3 mL of the samplesolution, add diluted methanol (1 in 2) to make exactly 50mL, and use this solution as the solution for system suitabil-ity test. Pipet 2 mL of the solution for system suitability test,and add diluted methanol (1 in 2) to make exactly 20 mL.Conˆrm that the peak area of josamycin obtained from 10mL of this solution is equivalent to 8 to 12z of that from 10mL of the solution for system suitability test.

System performance: Dissolve 0.05 g of Josamycin in 50mL of 0.1 mol/L potassium dihydrogen phosphate TS, pH2.0, and allow to stand at 409C for 3 hours. Adjust the pHof this solution to 6.8 to 7.2 with 2 mol/L sodium hydroxideTS, and add 50 mL of methanol. When the procedure is runwith 10 mL of this solution under the above operating condi-tions, the resolution between the peaks of josamycin S1,which relative retention time to josamycin is about 0.9, andjosamycin is not less than 2.0.

System repeatability: When the test is repeated 6 timeswith 10 mL of the solution for system suitability test underthe above operating conditions, the relative standard devia-


tion of the peak area of josamycin is not more than 1.5z.

Loss on drying Not more than 1.0z (0.5 g, in vacuum,phosphorus (V) oxide, 609C, 3 hours).



(1) Test organism—Bacillus subtilis ATCC 6633(2) Culture medium—Use the medium ii in 3) Medium

for other organisms under (1) Agar media for seed and baselayer. Adjust the pH of the medium so that it will be 7.9 to8.1 after sterilization.

(3) Standard solutions—Weigh accurately an amount ofJosamycin Reference Standard, equivalent to about 30 mg(potency), dissolve in 5 mL of methanol, add water to makeexactly 100 mL, and use this solution as the standard stocksolution. Keep the standard stock solution at 59C or below,and use within 7 days. Take exactly a suitable amount of thestandard stock solution before use, add water to make solu-tions so that each mL contains 30 mg (potency) and 7.5 mg(potency), and use these solutions as the high concentrationstandard solution and the low concentration standard solu-tion, respectively.

(4) Sample solutions—Weigh accurately an amount ofJosamycin, equivalent to about 30 mg (potency), dissolve in5 mL of methanol, and add water to make exactly 100 mL.Take exactly a suitable amount of this solution, add water tomake solutions so that each mL contains 30 mg (potency) and7.5 mg (potency), and use these solutions as the high concen-tration sample solution and the low concentration samplesolution, respectively.


Josamycin Propionateプロピオン酸ジョサマイシン


Josamycin Propionate contains not less than 843 mg(potency) per mg, calculated on the dried basis. Thepotency of Josamycin Propionate is expressed as mass(potency) of josamycin (C42H69NO15).

Description Josamycin Propionate occurs as a white tolight yellowish white crystalline powder.

It is very soluble in acetonitrile, freely soluble in methanoland in ethanol (99.5), and practically insoluble in water.

Identiˆcation (1) Determine the absorption spectrum ofa solution of Josamycin Propionate in methanol (1 in100,000) as directed under the Ultraviolet-visible Spec-trophotometry, and compare the spectrum with the Refer-ence Spectrum or the spectrum of a solution of Josamycin

Propionate Reference Standard prepared in the same man-ner as the sample solution: both spectra exhibit similar inten-sities of absorption at the same wavelengths.

(2) Dissolve 5 mg each of Josamycin Propionate andJosamycin Propionate Reference Standard in 50 mL ofdiluted acetonitrile (1 in 2), and use these solutions as thesample solution and the standard solution, respectively. Per-form the test with 10 mL each of the sample solution and thestandard solution as directed under the Liquid Chro-matography according to the following conditions: theretention time of the peak of josamycin propionate obtainedfrom the sample solution is the same with that of the peak ofjosamycin propionate from the standard solution.Operating conditions—

Detector, column, column temperature, mobile phase,and ‰ow rate: Proceed as directed in the operating condi-tions in the Purity (2).

Purity (1) Heavy metals—Proceed with 1.0 g of Josamy-cin Propionate according to Method 2, and perform the test.Prepare the control solution with 3.0 mL of Standard LeadSolution (not more than 30 ppm).

(2) Related substances—Dissolve 0.05 g of JosamycinPropionate in the mobile phase to make 50 mL, and use thissolution as the sample solution. Perform the test with 10 mLof the sample solution as directed under the Liquid Chro-matography according to the following conditions. Deter-mine each peak area by the automatic integration method,and calculate the amounts of each peak other than josamy-cin propionate by the area percentage method: the amountof any peak other than josamycin is not more than 6z, andthe total of these peaks is not more than 22z.Operating conditions—




Mobile phase: To 10 mL of triethylamine add water tomake 1000 mL, and adjust the pH to 4.3 with acetic acid(100). To 500 mL of this solution add 500 mL of acetoni-trile.

Flow rate: Adjust the ‰ow rate so that the retention timeof josamycin propionate is about 24 minutes.

Time span of measurement: About 3.5 times as long as theretention time of josamycin propionate after the solventpeak.System suitability—

Test for required detectability: Measure exactly 3 mL ofthe sample solution, add the mobile phase to make exactly50 mL, and use this solution as the solution for systemsuitability test. Measure exactly 2 mL of the solution for sys-tem suitability test, and add the mobile phase to make ex-actly 20 mL. Conˆrm that the peak area of josamycinpropionate obtained from 10 mL of this solution is equiva-lent to 8 to 12z of that from 10 mL of the solution for sys-


tem suitability test.System performance: Dissolve 5 mg of josamycin pro-

pionate and 2 mg of josamycin in 50 mL of the mobilephase. When the procedure is run with 10 mL of this solutionunder the above operating conditions, josamycin andjosamycin propionate are eluted in this order with the reso-lution between these peaks being not less than 25.

System repeatability: When the test is repeated 6 timeswith 10 mL of the solution for system suitability test underthe above operating conditions, the relative standard devia-tion of the peak area of josamycin propionate is not morethan 1.5z.

Loss on drying Not more than 1.0z (1 g, in vacuum,phosphorus (V) oxide, 609C, 3 hours).



(1) Test organism—Bacillus subtilis ATCC 6633(2) Culture medium—Use the medium ii in 3) Medium


(3) Standard solutions—Weigh accurately an amount ofJosamycin Propionate Reference Standard, equivalent toabout 20 mg (potency), dissolve in 10 mL of methanol, add1/15 mol/L phosphate buŠer solution, pH 5.6 to make ex-actly 50 mL, and use this solution as the standard stock solu-tion. Keep the standard stock solution at 59C or below, anduse within 3 days. Take exactly a suitable amount of thestandard stock solution before use, add 1/15 mol/L phos-phate buŠer solution, pH 5.6 to make solutions so that eachmL contains 80 mg (potency) and 20 mg (potency), and usethese solutions as the high concentration standard solutionand the low concentration standard solution, respectively.

(4) Sample solutions—Weigh accurately an amount ofJosamycin Propionate, equivalent to about 20 mg (potency),dissolve in 10 mL of methanol, add 1/15 mol/L phosphatebuŠer solution, pH 5.6 to make exactly 50 mL. Take exactlya suitable amount of this solution, add 1/15 mol/L phos-phate buŠer solution, pH 5.6 to make solutions so that eachmL contains 80 mg (potency) and 20 mg (potency), and usethese solutions as the high concentration sample solutionand the low concentration sample solution, respectively.


Add the following:

Kanamycin Monosulfate一硫酸カナマイシン

C18H36N4O11.H2SO4: 582.58O-3-Amino-3-deoxy-a-D-glucopyranosyl-(1→6)-O-[6-amino-6-deoxy-a-D-glucopyranosyl-(1→4)]-2-deoxy-D-streptamine monosulfate [25389-94-0 ]

Kanamycin Monosulfate contains not less than 750mg (potency) per mg, calculated on the dried basis. Thepotency of Kanamycin Monosulfate is expressed asmass (potency) of kanamycin (C18H36N4O11: 484.50).

Description Kanamycin Monosulfate occurs as a whitecrystalline powder.


Identiˆcation (1) Dissolve 50 mg of KanamycinMonosulfate in 3 mL of water, and add 6 mL of anthroneTS: a blue-purple color develops.

(2) Dissolve 20 mg each of Kanamycin Monosulfate andKanamycin Monosulfate Reference Standard in 1 mL ofwater, and use these solutions as the sample solution and thestandard solution. Perform the test with these solutions asdirected under the Thin-layer Chromatography. Spot 5 mLeach of the sample solution and the standard solution on aplate of silica gel for thin-layer chromatography. Developthe plate with the supernatant layer of a mixture of chlo-roform, ammonia solution (28) and methanol (2:1:1) to adistance of about 10 cm, and air-dry the plate. Spray evenlya solution of 0.2z ninhydrin-water saturated 1-butanol TSon the plate, and heat at 1009C for 10 minutes: the principalspots obtained from the sample solution and the standardsolution show a purple-brown color and the same Rf value.

(3) To a solution of Kanamycin Monosulfate (1 in 5) add1 drop of barium chloride TS: a white precipitate is formed.



Sulfuric acid Weigh accurately about 0.25 g of KanamycinMonosulfate, dissolve in 100 mL of water, adjust the pH to11.0 with ammonia solution (28), add exactly 10 mL of0.1 mol/L barium chloride VS, and titrate with 0.1 mol/L


disodium dihydrogen ethylenediamine tetraacetate VS untilthe color of the solution, blue-purple, disappears (indicator:0.5 mg of phthalein purple). At a near of the end-point add50 mL of ethanol (99.5). Perform a blank determination inthe same manner. The amount of sulfuric acid (SO4) is notless than 15.0z and not more than 17.0z, calculated on thedried basis.

Each mL of 0.1 mol/L barium chloride VS＝ 9.606 mg of SO4

Purity (1) Heavy metals—Proceed with 2.0 g ofKanamycin Monosulfate according to Method 4, and per-form the test. Prepare the control solution with 2.0 mL ofStandard Lead Solution (not more than 10 ppm).

(2) Arsenic—Prepare the test solution with 2.0 g ofKanamycin Monosulfate according to Method 4, and per-form the test (not more than 1 ppm).

(3) Related substances—Dissolve 0.30 g of KanamycinMonosulfate in water to make exactly 10 mL, and use thissolution as the sample solution. Separately, dissolve 45 mgof Kanamycin Monosulfate Reference Standard in water tomake exactly 50 mL, and use this solution as the standardsolution. Perform the test with these solutions as directedunder the Thin-layer Chromatography. Spot 1 mL each ofthe sample solution and the standard solution on a plate ofsilica gel for thin-layer chromatography. Develop the platewith a solution of potassium dihydrogen phosphate (3 in 40)to a distance of about 10 cm, and air-dry the plate. Sprayevenly a solution of ninhydrin in 1-butanol (1 in 100) on theplate, and heat at 1109C for 10 minutes: the spot other thanthe principal spot obtained from the sample solution is notmore intense than the spot from the standard solution.






(3) Standard solutions—Weigh accurately an amount ofKanamycin Monosulfate Reference Standard, previouslydried, equivalent to about 20 mg (potency), dissolve in dilut-ed phosphate buŠer solution, pH 6.0 (1 in 2) to make exactly50 mL, and use this solution as the standard stock solution.Keep the standard stock solution between 5 and 159C anduse within 30 days. Take exactly a suitable amount of thestandard stock solution before use, add 0.1 mol/L phos-phate buŠer solution, pH 8.0 to make solutions so that eachmL contains 20 mg (potency) and 5 mg (potency), and usethese solutions as the high concentration standard solutionand the low concentration standard solution, respectively.

(4) Sample solutions—Weigh accurately an amount ofKanamycin Monosulfate, equivalent to about 20 mg (poten-

cy), and dissolve in water to make exactly 50 mL. Take ex-actly a suitable amount of this solution, add 0.1 mol/Lphosphate buŠer solution, pH 8.0 to make solutions so thateach mL contains 20 mg (potency) and 5 mg (potency), anduse these solutions as the high concentration sample solutionand the low concentration sample solution, respectively.


Kanamycin Sulfate硫酸カナマイシン


Kanamycin Sulfate contains not less than 690 mg(potency) per mg, calculated on the dried basis. Thepotency of Kanamycin Sulfate is expressed as mass(potency) of kanamycin (C18H36N4O11: 484.50).

Description Kanamycin Sulfate occurs as a white to yel-lowish white powder.


Identiˆcation (1) Dissolve 20 mg each of Kanamycin Sul-fate and Kanamycin Monosulfate Reference Standard in 1mL of water, and use these solutions as the sample solutionand the standard solution. Perform the test with these solu-tions as directed under the Thin-layer Chromatography.Spot 5 mL each of the sample solution and the standard solu-tion on a plate of silica gel for thin-layer chromatography.Develop the plate with a mixture of chloroform, ammoniasolution (28) and methanol (2:1:1) to a distance of about 10cm, and air-dry the plate. Spray evenly 0.2z ninhydrin-water saturated 1-butanol TS on the plate, and heat at 1009Cfor 10 minutes: the principal spots obtained from the samplesolution and the standard solution show a purple-browncolor and the same Rf value.

(2) A solution of Kanamycin Sulfate (1 in 10) respondsto the Qualitative Test (1) for sulfate.



pH The pH of a solution obtained by dissolving 1.0 g ofKanamycin Sulfate in 20 mL of water is between 6.0 and 7.5.

Purity (1) Clarity and color of solution—Dissolve 1.5 gof Kanamycin Sulfate in 5 mL of water: the solution is clearand colorless to pale yellow.

(2) Heavy metals—Proceed with 1.0 g of KanamycinSulfate according to Method 4, and perform the test. Pre-pare the control solution with 3.0 mL of Standard Lead So-lution (not more than 30 ppm).

(3) Arsenic—Prepare the test solution with 2.0 g ofKanamycin Sulfate according to Method 3, and perform thetest (not more than 1 ppm).


(4) Related substances—Dissolve 0.30 g of KanamycinSulfate in water to make exactly 10 mL, and use this solutionas the sample solution. Separately, dissolve 9.0 mg ofKanamycin Monosulfate Reference Standard in water tomake exactly 10 mL, and use this solution as the standardsolution. Perform the test with these solutions as directedunder the Thin-layer Chromatography. Spot 1 mL each ofthe sample solution and the standard solution on a plate ofsilica gel for thin-layer chromatography. Develop the platewith a solution of potassium dihydrogen phosphate (3 in 40)to a distance of about 10 cm, and air-dry the plate. Sprayevenly a solution of ninhydrin in 1-butanol (1 in 100) on theplate, and heat at 1109C for 10 minutes: the spot other thanthe principal spot obtained from the sample solution is notmore intense than the spot from the standard solution.





(3) Standard solutions—Weigh accurately an amount ofKanamycin Monosulfate Reference Standard, previously d-ried, equivalent to about 20 mg (potency), dissolve in dilutedphosphate buŠer solution, pH 6.0 (1 in 2) to make exactly 50mL, and use this solution as the standard stock solution.Keep the standard stock solution at 5 to 159C and use within30 days. Take exactly a suitable amount of the standardstock solution before use, add 0.1 mol/L phosphate buŠersolution, pH 8.0 to make solutions so that each mL contains20 mg (potency) and 5 mg (potency), and use these solutionsas the high concentration standard solution and the low con-centration standard solution, respectively.

(4) Sample solutions—Weigh accurately an amount ofKanamycin Sulfate, equivalent to about 20 mg (potency),and dissolve in water to make exactly 50 mL. Take exactly asuitable amount of this solution, add 0.1 mol/L phosphatebuŠer solution, pH 8.0 to make solutions so that each mLcontains 20 mg (potency) and 5 mg (potency), and use thesesolutions as the high concentration sample solution and thelow concentration sample solution, respectively.


Add the following:

Ketotifen Fumarateフマル酸ケトチフェン

C19H19NOS.C4H4O4: 425.504-(1-Methylpiperidin-4-ylidene)-4H-benzo[4,5]cyclohepta[1,2-b]thiophen-10(9H )-onemonofumarate [34580-14-8 ]

Ketotifen Fumarate, when dried, contains not lessthan 99.0z and not more than 101.0z of ketotifenfumarate (C19H19NOS.C4H4O4).

Description Ketotifen Fumarate occurs as a white to lightyellowish white crystalline powder.

It is sparingly soluble in methanol and in acetic acid (100),and slightly soluble in water, in ethanol (99.5) and in aceticanhydride.

Melting point: about 1909C (with decomposition).

Identiˆcation (1) Prepare the test solution with 0.03 g ofKetotifen Fumarate as directed under the Oxygen FlaskCombustion Method using 20 mL of water as the absorbingliquid: the test solution responds to the Qualitative Tests forsulfate.

(2) Determine the absorption spectrum of a solution ofKetotifen Fumarate in methanol (1 in 50,000) as directed un-der the Ultraviolet-visible Spectrophotometry, and comparethe spectrum with the Reference Spectrum: both spectra ex-hibit similar intensities of absorption at the samewavelengths.

(3) Determine the infrared absorption spectrum ofKetotifen Fumarate, previously dried, as directed in thepotassium bromide disk method under the Infrared Spec-trophotometry, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of ab-sorption at the same wave numbers.

Purity (1) Chloride—Dissolve 0.6 g of KetotifenFumarate in 2.5 mL of sodium carbonate TS in a crucible,heat on a water bath to dryness, and ignite at about 5009C.Dissolve the residue in 15 mL of water, ˆlter if necessary,neutralize with diluted nitric acid (3 in 10), and add 6 mL ofdilute nitric acid and water to make 50 mL. Perform the testusing this solution as the test solution. Prepare the controlsolution as follows: To 0.25 mL of 0.01 mol/L hydrochloricacid VS add 2.5 mL of sodium carbonate TS, the usedamount of diluted nitric acid (3 in 10) for the neutralization,6 mL of dilute nitric acid and water to make 50 mL (notmore than 0.015z).


(2) Heavy metals—Proceed with 1.0 g of KetotifenFumarate according to Method 2, and perform the test. Pre-pare the control solution with 2.0 mL of Standard Lead So-lution (not more than 20 ppm).

(3) Related substances—Dissolve 0.10 g of KetotifenFumarate in 10 mL of a mixture of methanol and ammoniaTS (99:1), and use this solution as the sample solution. Pipet1 mL of the sample solution, and add a mixture of methanoland ammonia TS (99:1) to make exactly 25 mL. Pipet 1 mLof this solution, add a mixture of methanol and ammonia TS(99:1) to make exactly 20 mL, and use this solution as thestandard solution. Perform the test with these solutions asdirected under the Thin-layer Chromatography. Spot 10 mLeach of the sample solution and the standard solution on aplate of silica gel for thin-layer chromatography. Developthe plate with a mixture of acetonitrile, water and ammoniasolution (28) (90:10:1) to a distance of about 15 cm, and air-dry the plate. Spray evenly DragendorŠ's TS for sprayingand then hydrogen peroxide TS on the plate: the number ofthe spot other than the principal spot obtained from thesample solution is not more than four, and they are not moreintense than the spot from the standard solution.



Assay Weigh accurately about 0.35 g of KetotifenFumarate, previously dried, dissolve in 80 mL of a mixtureof acetic anhydride and acetic acid (100) (7:3), and titratewith 0.1 mol/L perchloric acid VS (potentiometric titra-tion). Perform a blank determination, and make any neces-sary correction.

Each mL of 0.1 mol/L perchloric acid VS＝ 42.55 mg of C19H19NOS.C4H4O4


Kitasamycinキタサマイシン

Change the Content ratio of the active principleto read:

Content ratio of the active principle Dissolve 0.02 g ofKitasamycin in diluted acetonitrile (1 in 2) to make 20 mL,and use this solution as the sample solution. Perform the testwith 5 mL of the sample solution as directed under the LiquidChromatography according to the following conditions, andmeasure each peak area by the automatic integrationmethod. Calculate the amounts of leucomycin A5, leucomy-cin A4 and leucomycin A1 by the area percentage method:the amounts of leucomycin A5, leucomycin A4 and leucomy-cin A1 are 40 to 70z, 5 to 25z and 3 to 12z, respectively.Relative retention times of leucomycin A4 and leucomycinA1 to that of leucomycin A5 are 1.2 and 1.5, respectively.



ameter and 15 cm in length, packed with octylsilanized silicagel for liquid chromatography (5 mm in particle diameter).


Mobile phase: To a volume of a solution of ammoniumacetate (77 in 500) add diluted phosphoric acid (1 in 150) toadjust to pH 5.5. To 370 mL of this solution add 580 mL ofmethanol and 50 mL of acetonitrile.

Flow rate: Adjust the ‰ow rate so that the retention timeof leucomycin A5 is about 8 minutes.

Time span of measurement: About 3 times as long as theretention time of leucomycin A5.System suitability—

System performance: Dissolve about 20 mg each of Leu-comycin A5 Reference Standard and Josamycin ReferenceStandard in 20 mL of diluted acetonitrile (1 in 2). When theprocedure is run with 5 mL of this solution under the aboveoperating conditions, leucomycin A5 and josamycin are elut-ed in this order with the resolution between these peaksbeing not less than 5.

System repeatability: When the test is repeated 6 timeswith 5 mL of the sample solution under the above operatingconditions, the relative standard deviation of the peak areaof leucomycin A5 is not more than 1.0z.


Assay(3) Standard solutions—Weigh accurately an amount of

Leucomycin A5 Reference Standard equivalent to about30 mg (potency), dissolve in 10 mL of methanol, add waterto make exactly 100 mL, and use this solution as the stan-dard stock solution. Keep the standard stock solution at 59Cor below and use within 3 days. Take exactly a suitableamount of the standard stock solution before use, add phos-phate buŠer solution, pH 8.0 to make solutions so that eachmL contains 30 mg (potency) and 7.5 mg (potency), and usethese solutions as the high concentration standard solutionand the low concentration standard solution, respectively.


Add the following:

Kitasamycin TartrateLeucomycin Tartrate

酒石酸キタサマイシン

(Leucomycin A1, A5, A7, A9, A13 tartrate)(3R,4R,5S,6R,8R,9R,10E,12E,15R )-5-[O-(4-O-Acyl-2,6-dideoxy-3-C-methyl-a-L-ribo-hexopyranosyl)-(1→4)-3,6-dideoxy-3-dimethylamino-b-D-glucopyranosyloxy]-6-formylmethyl-3,9-dihydroxy-4-methoxy-8-methylhexadeca-10,12-dien-15-olide mono-(2R,3R )-tartrateLeucomycin A1 tartrate: acyl＝ 3-methylbutanoylLeucomycin A5 tartrate: acyl＝ butanoylLeucomycin A7 tartrate: acyl＝ propanoylLeucomycin A9 tartrate: acyl＝ acetylLeucomycin A13 tartrate: acyl＝ hexanoyl

(Leucomycin A3, A4, A6, A8 tartrate)(3R,4R,5S,6R,8R,9R,10E,12E,15R )-3-Acetoxy-5-[O-(4-O-acyl-2,6-dideoxy-3-C-methyl-a-L-ribo-hexopyranosyl)-(1→4)-3,6-dideoxy-3-dimethylamino-b-D-glucopyranosyloxy]-6-formylmethyl-9-hydroxy-4-methoxy-8-methylhexadeca-10,12-dien-15-olidemono-(2R,3R )-tartrateLeucomycin A3 tartrate: acyl＝ 3-methylbutanoylLeucomycin A4 tartrate: acyl＝ butanoylLeucomycin A6 tartrate: acyl＝ propanoylLeucomycin A8 tartrate: acyl＝ acetyl[37280-56-1 ]

Kitasamycin Tartrate contains not less than 1300 mg(potency) per mg, calculated on the anhydrous basis.The potency of Kitasamycin Tartrate is expressed asmass (potency) of kitasamycin based on the amount ofleucomycin A5 (C39H65NO14: 771.93). 1 mg (potency)of Kitasamycin Tartrate is equivalent to 0.530 mg ofleucomycin A5 (C39H65NO14).

Description Kitasamycin Tartrate occurs as a white to lightyellowish white powder.

It is very soluble in water, in methanol and in ethanol(99.5).

Identiˆcation (1) Determine the absorption spectrum ofa solution of Kitasamycin Tartrate in methanol (1 in 40,000)as directed under the Ultraviolet-visible Spectrophotometry,and compare the spectrum with the Reference Spectrum:both spectra exhibit similar intensities of absorption at thesame wavelengths.

(2) Determine the infrared absorption spectrum ofKitasamycin Tartrate as directed in the potassium bromidedisk method under the Infrared Spectrophotometry, andcompare the spectrum with the Reference Spectrum: bothspectra exhibit similar intensities of absorption at the samewave numbers.

(3) Dissolve 1 g of Kitasamycin Tartrate in 20 mL ofwater, add 3 mL of sodium hydroxide TS, add 20 mL of n-butyl acetate, shake well, and discard the n-butyl acetate lay-er. To the aqueous layer add 20 mL of n-butyl acetate, andshake well. The aqueous layer so obtained responds to theQualitative Test (1) for tartrate.

pH Dissolve 3.0 g of Kitasamycin Tartrate in 100 mL ofwater: the pH of the solution is between 3.0 and 5.0.

Content ratio of the active principle Dissolve 20 mg ofKitasamycin Tartrate in diluted acetonitrile (1 in 2) to make20 mL, and use this solution as the sample solution. Performthe test with 5 mL of the sample solution as directed underthe Liquid Chromatography according to the following con-ditions, determine the peak areas by the automatic integra-tion method, and calculate the amounts of leucomycin A5,leucomycin A4 and leucomycin A1 by the area percentagemethod: the amount of leucomycin A5 is 40 – 70z, leu-comycin A4 is 5 – 25z, and leucomycin A1 is 3 – 12z. Therelative retention times of leucomycin A4 and leucomycin A1

with respect to leucomycin A5 are 1.2 and 1.5, respectively.Operating conditions—




Mobile phase: To a suitable amount of a solution of am-monium acetate (77 in 500) add diluted phosphoric acid (1 in150) to adjust the pH to 5.5. To 370 mL of this solution add580 mL of methanol and 50 mL of acetonitrile.


Flow rate: Adjust the ‰ow rate so that the retention timeof leucomycin A5 is about 8 minutes.

Time span of measurement: About 3 times as long as theretention time of leucomycin A5.System suitability—

System performance: Dissolve about 20 mg of Leucomy-cin A5 Reference Standard and about 20 mg of JosamycinReference Standard in 20 mL of diluted acetonitrile (1 in 2).When the procedure is run with 5 mL of this solution underthe above operating conditions, leucomycin A5 and josamy-cin are eluted in this order with the resolution between thesepeaks being not less than 5.

System repeatability: When the test is repeated 6 timeswith 5 mL of the sample solution under the above operatingconditions, the relative standard deviation of the peak areaof leucomycin A5 is not more than 1.0z.

Purity (1) Clarity and color of solution—Dissolve 1.0 gof Kitasamycin Tartrate in 10 mL of water: the solution isclear and colorless or light yellow.

(2) Heavy metals—Proceed with 1.0 g of KitasamycinTartrate according to Method 2, and perform the test. Pre-pare the control solution with 3.0 mL of Standard Lead So-lution (not more than 30 ppm).





(3) Standard solutions—Weigh accurately an amount ofLeucomycin A5 Reference Standard, equivalent to about 30mg (potency), dissolve in 10 mL of methanol, add water tomake exactly 100 mL, and use this solution as the standardstock solution. Keep the standard stock solution at not ex-ceeding 59C, and use within 3 days. Take exactly a suitableamount of the standard stock solution before use, add phos-phate buŠer solution, pH 8.0 to make solutions so that eachmL contains 30 mg (potency) and 7.5 mg (potency), and usethese solutions as the high concentration standard solutionand the low concentration standard solution, respectively.

(4) Sample solutions—Weigh accurately an amount ofKitasamycin Tartrate, equivalent to about 30 mg (potency),and dissolve in water to make exactly 100 mL. Take exactly asuitable amount of this solution, add phosphate buŠer solu-tion, pH 8.0 to make solutions so that each mL contains 30mg (potency) and 7.5 mg (potency), and use these solutions asthe high concentration sample solution and the low concen-tration sample solution, respectively.


Latamoxef Sodiumラタモキセフナトリウム


Latamoxef Sodium contains not less than 830 mg(potency) per mg, calculated on the anhydrous basis.The potency of Latamoxef Sodium is expressed asmass (potency) of latamoxef (C20H20N6O9S: 520.47).

Description Latamoxef Sodium occurs as white to lightyellowish white, powder or masses.

It is very soluble in water, freely soluble in methanol, andslightly soluble in ethanol (95).

Identiˆcation (1) Determine the absorption spectrum ofa solution of Latamoxef Sodium (3 in 100,000) as directedunder the Ultraviolet-visible Spectrophotometry, and com-pare the spectrum with the Reference Spectrum: both spec-tra exhibit similar intensities of absorption at the samewavelengths.

(2) Determine the infrared absorption spectrum ofLatamoxef Sodium as directed in the potassium bromidedisk method under the Infrared Spectrophotometry, andcompare the spectrum with the Reference Spectrum: bothspectra exhibit similar intensities of absorption at the samewave numbers.

(3) Determine the spectrum of a solution of LatamoxefSodium in heavy water for nuclear magnetic resonance spec-troscopy (1 in 10) as directed under the Nuclear MagneticResonance Spectroscopy (1H), using sodium 3-trimethylsilyl-propanesulfonate for nuclear magnetic resonance spec-troscopy as an internal reference compound: it exhibits sin-gle signals, A and B, at around d3.5 ppm and at aroundd4.0 ppm. The ratio of the integrated intensity of these sig-nals, A:B, is about 1:1.

(4) Latamoxef Sodium responds to the Qualitative Test(1) for sodium salt.

Optical rotation [a]D20: －32 –－409(0.5 g calculated onthe anhydrous basis, phosphate buŠer solutiuon, pH 7.0, 50mL, 100 mm).

pH The pH of a solution obtained by dissolving 1.0 g ofLatamoxef Sodium in 10 mL of water is between 5.0 and7.0.

Purity (1) Clarity and color of solution—Dissolve 1.0 gof Latamoxef Sodium in 10 mL of water: the solution isclear and pale yellow.

(2) Heavy metals—Carbonize 1.0 g of Latamoxef Sodi-um by heating gently, previously powdered if it is masses.After cooling, add 10 mL of a solution of magnesium nitratehexahydrate in ethanol (1 in 10), and burn the ethanol. Aftercooling, add 1 mL of sulfuric acid. Proceed according toMethod 4, and perform the test. Prepare the control solutionwith 2.0 mL of Standard Lead Solution (not more than 20ppm).


(3) Arsenic—Prepare the test solution by dissolving1.0 g of Latamoxef Sodium in 20 mL of water, and performthe test (not more than 2 ppm).

(4) Related substances—Dissolve an amount ofLatamoxef Sodium, equivalent to about 25 mg (potency), inwater to make exactly 50 mL, and use this solution as thesample solution. Pipet 2 mL of the sample solution, addwater to make exactly 100 mL, and use this solution as thestandard solution. Perform the test with exactly 5 mL each ofthe sample solution and the standard solution as directed un-der the Liquid Chromatography according to the followingconditions, and determine each peak area by the automaticintegration method: the peak area of 1-methyl-1H-tetrazole-5-thiol, having the relative retention time of about 0.5 withrespect to the ˆrst eluted peak of the two peaks of latamox-ef, obtained from the sample solution is not more than thepeak area of latamoxef from the standard solution, and thepeak area of decarboxylatamoxef, having the relative reten-tion time of about 1.7 with respect to the ˆrst peak of thetwo peaks of latamoxef, is not more than 2 times that oflatamoxef from the standard solution. For this calculation,use the peak area for 1-methyl-1H-tetrazole-5-thiol aftermultiplying by its response factor, 0.52.Operating conditions—

Proceed as directed in the operating conditions in theAssay.System suitability—


System repeatability: When the test is repeated 6 timeswith 5 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peakarea of latamoxef is not more than 2.0z.


Isomer ratio Dissolve 25 mg of Latamoxef Sodium inwater to make 50 mL, and use this solution as the sample so-lution. Perform the test with 5 mL of the sample solution asdirected under the Liquid Chromatography according to thefollowing conditions, and determine the areas, Aa and Ab, ofthe two peaks in order of elution, which appear close to eachother at the retention time of about 10 minutes: Aa/Ab is be-tween 0.8 and 1.4.Operating conditions—




Mobile phase: Dissolve 7.7 g of ammonium acetate inwater to make 1000 mL. To 950 mL of this solution add50 mL of methanol.

Flow rate: Adjust the ‰ow rate so that the retention timeof the ˆrst eluted peak of latamoxef is about 8 minutes.


of the sample solution under the above operating conditions,the resolution between the two peaks of latamoxef is not lessthan 3.

System repeatability: When the test is repeated 3 timeswith 5 mL of the sample solution under the above operatingconditions, the relative standard deviation of the area of theˆrst eluted peak of latamoxef is not more than 2.0z.

Assay Weigh accurately an amount of Latamoxef Sodiumand Latamoxef Ammonium Reference Standard, equivalentto about 25 mg (potency) each, dissolve in exactly 5 mL ofthe internal standard solution, add water to make 50 mL,and use these solutions as the sample solution and the stan-dard solution. Perform the test with 5 mL each of the samplesolution and the standard solution as directed under the Liq-uid Chromatography according to the following conditions,and determine the ratios, QT and QS, of the peak area oflatamoxef to that of the internal standard.

Amount [mg (potency)] of latamoxef (C20H20N6O9S)

＝WS×QT

QS× 1000

WS: Amount [mg (potency)] of Latamoxef AmmoniumReference Standard

Internal standard solution—A solution of m-cresol (3 in200).Operating conditions—




Mobile phase: Dissolve 6.94 g of potassium dihydrogenphosphate, 3.22 g of disodium hydrogen phosphate 12-waterand 1.60 g of tetra n-butylammonium bromide in water tomake exactly 1000 mL. To 750 mL of this solution add250 mL of methanol.

Flow rate: Adjust the ‰ow rate so that the retention timeof latamoxef is about 7 minutes.System suitability—

System performance: When the procedure is run with 5 mLof the standard solution under the above operating condi-tions, latamoxef and the internal standard are eluted in thisorder with the resolution between these peaks being not lessthan 5.

System repeatability: When the test is repeated 6 timeswith 5 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratiosof the peak area of latamoxef to that of the internal standardis not more than 1.0z.

Containers and storage Containers—Tight containers.Storage—Not exceeding 59C.


Add the following:

Lenampicillin Hydrochloride塩酸レナンピシリン

C21H23N3O7S.HCl: 497.955-Methyl-2-oxo[1,3]dioxol-4-ylmethyl (2S,5R,6R )-6-[(2R )-2-amino-2-phenylacetylamino]-3,3-dimethyl-7-oxo-4-thia-1-azabicyclo[3.2.0]heptane-2-carboxylatemonohydrochloride [80734-02-7 ]

Lenampicillin Hydrochloride is the hydrochloride ofampicillin methyloxodioxolenylmethyl ester.

It contains not less than 653 mg (potency) per mg,calculated on the anhydrous basis and corrected by theamount of the residual solvents. The potency ofLenampicillin Hydrochloride is expressed as mass(potency) of ampicillin (C16H19N3O4S: 349.40).

Description Lenampicillin Hydrochloride occurs as a whiteto light yellowish white powder.

It is very soluble in water, in methanol and in ethanol (95),and freely soluble in N,N-dimethylformamide.

Identiˆcation (1) Determine the infrared absorptionspectrum of Lenampicillin Hydrochloride as directed in thepotassium chloride disk method under the Infrared Spec-trophotometry, and compare the spectrum with the Refer-ence Spectrum or the spectrum of Lenampicillin Hydrochlo-ride Reference Standard: both spectra exhibit similar intensi-ties of absorption at the same wave numbers.

(2) To 1 mL of a solution of Lenampicillin Hydrochlo-ride (1 in 100) add 0.5 mL of dilute nitric acid and 1 drop ofsilver nitrate TS: a white precipitate is formed.


the anhydrous de-residual solventization basis, ethanol (95),20 mL, 100 mm).

Purity (1) Heavy metals—Proceed with 2.0 g of Lenam-picillin Hydrochloride according to Method 2, and performthe test. Prepare the control solution with 2.0 mL of Stan-dard Lead Solution (not more than 10 ppm).

(2) Arsenic—Prepare the test solution with 1.0 g ofLenampicillin Hydrochloride according to Method 3, andperform the test (not more than 2 ppm).

(3) Free ampicillin—Weigh accurately about 0.1 g ofLenampicillin Hydrochloridein, dissolve in exactly 10 mL ofthe internal standard solution, and use this solution as thesample solution. Separately, weigh accurately an amount ofAmpicillin Reference Standard, equivalent to about 25 mg(potency), and dissolve in water to make exactly 100 mL.Pipet 2 mL of this solution, add exactly 10 mL of the inter-nal standard solution, and use this solution as the standard

solution. The sample solution should be used to the follow-ing test immediately after the solution is prepared. Performthe test with 10 mL each of the sample solution and the stan-dard solution as directed under the Liquid Chromatographyaccording to the following conditions, and determine the ra-tios, QT and QS, of the peak height of ampicillin to that ofthe internal standard: the amount of ampicillin is not morethan 1.0z.

Amount (z) of ampicillin (C16H19N3O4S)

＝WS

WT×

QT

QS× 2

WS: Amount [mg (potency)] of Ampicillin ReferenceStandard

WT: Amount (mg) of the sample

Internal standard solution—A solution of anhydrouscaŠeine in the mobile phase (1 in 50,000).Operating conditions—




Mobile phase: Dissolve 1.22 g of potassium dihydrogenphosphate in water to make 900 mL, and add 100 mL ofacetonitrile.


System performance: When the procedure is run with 10mL of the standard solution under the above operating con-ditions, ampicillin and the internal standard are eluted inthis order with the resolution between these peaks being notless than 5.

System repeatability: When the test is repeated 6 timeswith 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratiosof the peak height of ampicillin to that of the internal stan-dard is not more than 5z.

(4) Penicilloic acid—Weigh accurately about 0.1 g ofLenampicillin Hydrochloride, dissolve in water to make ex-actly 100 mL, and use this solution as the sample solution.Pipet 10 mL of the sample solution, add 10 mL of potassiumhydrogen phthalate buŠer solution, pH 4.6 and exactly 10mL of 0.005 mol/L iodine VS, allow to stand for exactly 15minutes while protecting from exposure to light, and titratewith 0.01 mol/L sodium thiosulfate VS (indicator: 1 mL ofstarch TS). Perform a blank determination, and make anynecessary correction: the amount of penicilloic acid(C16H21N3O5S: 367.42) is not more than 3.0z.

Each mL of 0.01 mol/L sodium thiosulfate VS＝ 0.45 mg of C16H21N3O5S

(5) Residual solvent—Weigh accurately about 0.25 g ofLenampicillin Hydrochloride, dissolve in exactly 1 mL of


the internal standard solution, add N,N-dimethylformamideto make 5 mL, and use this solution as the sample solution.Separately, weigh accurately about 80 mg of 2-propanol andabout 0.12 g of ethyl acetate, and add N,N-dimethylfor-mamide to make exactly 100 mL. Pipet 1 mL and 3 mL ofthis solution, add exactly 1 mL each of the internal standardsolution, add N,N-dimethylformamide to make 5 mL, anduse these solutions as the standard solution (1) and the stan-dard solution (2), respectively. Perform the test with 4 mLeach of the sample solution, the standard solution (1) andthe standard solution (2) as directed under the Gas Chro-matography according to the following conditions, and de-termine the ratios, QTa and QTb, of the peak height of 2-propanol and ethyl acetate to that of the internal standard ofthe sample solution, the ratios, QSa1 and QSb1, of the peakheight of 2-propanol and ethyl acetate to that of the internalstandard of the standard solution (1) and the ratios, QSa2 andQSb2, of the peak height of 2-propanol and ethyl acetate tothat of the internal standard of the standard solution (2).Calculate the amounts of 2-propanol and ethyl acetate bythe following equations: not more than 0.7z and not morethan 1.7z, respectively.

Amount (z) of 2-propanol

＝WSa

WT×

2QTa－ 3QSa1＋ QSa2

QSa2－ QSa1

Amount (z) of ethyl acetate

＝WSb

WT×

2QTb－ 3QSb1＋ QSb2

QSb2－ QSb1

WSa: Amount (g) of 2-propanolWSb: Amount (g) of ethyl acetateWT: Amount (g) of the sample

Internal standard solution—A solution of cyclohexane inN,N-dimethylformamide (1 in 1000).Operating conditions—

Detector: A hydrogen ‰ame-ionization detector.Column: A glass column 3 mm in inside diameter and 3 m

in length, packed with siliceous earth for gas chromato-graphy (180 – 250 mm in particle diameter) coated withtetrakishydroxypropylethylenediamine for gas chromato-graphy at the ratio of 10 to 15z.


Injection port temperature: A constant temperature ofabout 1609C.


of the internal standard is about 1 minute.System suitability—

System performance: When the procedure is run with 4 mLof the standard solution (2) under the above operating con-ditions, the internal standard, ethyl acetate and 2-propanolare eluted in this order, and the resolution between the peaksof the internal standard and ethyl acetate is not less than 2.0.

System repeatability: When the test is repeated 3 timeswith 4 mL of the standard solution (2) under the above oper-ating conditions, the relative standard deviation of the ratios

of the peak height of ethyl acetate to that of the internalstandard is not more than 5z.



Assay Weigh accurately an amount of LenampicillinHydrochloride and Lenampicillin Hydrochloride ReferenceStandard, equivalent to about 0.1 g (potency), dissolve eachin the internal standard solution to make exactly 10 mL, anduse these solutions as the sample solution and the standardsolution. Perform the test with 5 mL each of the sample solu-tion and the standard solution as directed under the LiquidChromatography according to the following conditions, anddetermine the ratios, QT and QS, of the peak area of lenam-picillin to that of the internal standard.


＝WS×QT

QS× 1000

WS: Amount [mg (potency)] of Lenampicillin Hydrochlo-ride Reference Standard

Internal standard solution—A solution of ethyl aminoben-zoate in the mobile phase (1 in 4000).Operating conditions—




Mobile phase: Dissolve 9.53 g of potassium dihydrogenphosphate in water to make exactly 700 mL, and addacetonitrile to make exactly 1000 mL.

Flow rate: Adjust the ‰ow rate so that the retention timeof lenampicillin is about 6 minutes.System suitability—

System performance: When the procedure is run with 5 mLof the standard solution under the above operating condi-tions, lenampicillin and the internal standard are eluted inthis order with the resolution between these peaks being notless than 10.

System repeatability: When the test is repeated 6 timeswith 5 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratiosof the peak area of lenampicillin to that of the internal stan-dard is not more than 1.0z.



Lincomycin Hydrochloride塩酸リンコマイシン


Lincomycin Hydrochloride contains not less than825 mg (potency) per mg, calculated on the anhydrousbasis. The potency of Lincomycin Hydrochlorideis expressed as mass (potency) of lincomycin(C18H34N2O6S: 406.54).

Description Lincomycin Hydrochloride occurs as white,crystals or crystalline powder.

It is freely soluble in water and in methanol, sparinglysoluble in ethanol (95), and very slightly soluble in acetoni-trile.

Identiˆcation (1) Determine the infrared absorptionspectrum of Lincomycin Hydrochloride as directed in thepaste method under the Infrared Spectrophotometry, andcompare the spectrum with the Reference Spectrum or thespectrum of Lincomycin Hydrochloride Reference Stan-dard: both spectra exhibit similar intensities of absorption atthe same wave numbers.

(2) A solution of Lincomycin Hydrochloride (1 in 100)responds to the Qualitative Test (2) for chloride.

Optical rotation [a]D20: ＋135 –＋1509(0.5 g, water, 25mL, 100 mm).

pH Dissolve 0.10 g of Lincomycin Hydrochloride in 1 mLof water: 3.0 – 5.5.

Purity (1) Clarity and color of solution—Dissolve 1.0 gof Lincomycin Hydrochloride in 10 mL of water: the solu-tion is clear and colorless.

(2) Heavy metals—Proceed with 2.0 g of LincomycinHydrochloride according to Method 4, and perform the test.Prepare the control solution with 1.0 mL of Standard LeadSolution (not more than 5 ppm).

(3) Lincomycin B—Perform the test with 20 mL of thesample solution obtained in the Assay as directed under theLiquid Chromatography according to the following condi-tions, and determine the peak areas of lincomycin and lin-comycin B, having the relative retention time of about 0.5with respect to lincomycin, by the automatic integrationmethod: the peak area of lincomycin B is not more than5.0z of the sum of the peak areas of lincomycin and lin-comycin B.Operating conditions—


Test for required detectability: Measure exactly 1 mL ofthe sample solution, and add the mobile phase to make ex-actly 20 mL. Conˆrm that the peak area of lincomycin ob-tained from 20 mL of this solution is equivalent to 3.5 to

6.5z of that obtained from 20 mL of the sample solution.System performance, and system repeatability: Proceed as

directed in the system suitability in the Assay.


Assay Weigh accurately an amount of LincomycinHydrochloride and Lincomycin Hydrochloride ReferenceStandard, equivalent to about 10 mg (potency), dissolveeach in the mobile phase to make exactly 10 mL, and usethese solutions as the sample solution and the standard solu-tion. Perform the test with exactly 20 mL each of the samplesolution and the standard solution as directed under the Liq-uid Chromatography according to the following conditions,and determine the peak areas, AT and AS, of lincomycin.

Amount [mg (potency)] of lincomycin (C18H34N2O6S)

＝WS×AT

AS× 1000

WS: Amount [mg (potency)] of Lincomycin Hydrochlo-ride Reference Standard


(wavelength: 210 nm).Column: A stainless steel column 4 mm in inside diameter

and 25 cm in length, packed with octylsilanized silica gel forliquid chromatography (5 mm in particle diameter).


Mobile phase: To 13.5 mL phosphoric acid add water tomake 1000 mL, and adjust the pH to 6.0 with ammonia TS.To 780 mL of this solution add 150 mL of acetonitrile and150 mL of methanol.

Flow rate: Adjust the ‰ow rate so that the retention timeof lincomycin is about 9 minutes.System suitability—

System performance: When the procedure is run with 20mL of the standard solution under the above operating con-ditions, the theoretical plates and the symmetrical coe‹cientof the peak of lincomycin are not less than 4000 and notmore than 1.3, respectively.

System repeatability: When the test is repeated 6 timeswith 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peakarea of lincomycin is not more than 2.0z.


Liothyronine Sodium Tabletsリオチロニンナトリウム錠

Change the Content uniformity to read:

Content uniformity Place 1 tablet of Liothyronine SodiumTablets in a glass-stoppered centrifuge tube, add exactly 10mL of 0.01 molWL sodium hydroxide TS, warm at 509C for


15 minutes, and shake vigorously for 20 minutes. Centrifugefor 5 minutes, and ˆlter the supernatant liquid, if necessary.Pipet a deˆnite volume of this solution, and add a volume of0.01 molWL sodium hydroxide VS to prepare a deˆnitevolume of a solution containing about 0.5 mg of liothyroninesodium (C15H11I3NNaO4) per mL. Pipet 5 mL of this solu-tion, add exactly 1 mL of the internal standard solution, anduse this solution as the sample solution. Perform the testwith 200 mL of the sample solution as directed under the Liq-uid Chromatography according to the following conditions,and calculate the ratio of the peak area of the liothyronine tothat of the internal standard. Calculate the mean value ofthe ratios of each peak area of 10 samples: the deviation (z)of each ratio of the peak area from the mean value should benot more than 15z. When the deviation (z) is more than15z, and 1 sample shows not more than 25z, performanother test with 20 samples. Calculate the deviation (z) ofeach ratio of the peak area from the mean value of the 30samples used in the two tests: there should be not more than1 sample with the deviation more than 15z but not morethan 25z, and no sample should deviate by more than 25z.Internal standard solution—A solution of propylparahy-droxybenzoate in a mixture of methanol and diluted phos-phoric acid (1 in 10) (9:1) (1 in 250,000).Operating conditions—


Column: A stainless steel column 4.6 mm in inside di-ameter and 15 cm in length, packed with octadecylsylanizedsilica gel for liquid chromatography (5 mm in particle di-ameter).


Mobile phase: Diluted methanol (57 in 100).Flow rate: Adjust the ‰ow rate so that the retention time

of liothyronine is about 9 minutes.System suitability—

System performance: To 5 mL of a solution of liothyro-nine sodium in 0.01 mol/L sodium hydroxide TS (1 in2,000,000) add 1 mL of the internal standard solution, anduse this solution as the solution for system suitability test.When the procedure is run with 200 mL of this solution un-der the above operating conditions, the internal standardand liothyronine are eluted in this order with the resolutionbetween these peaks being not less than 2.0.

System repeatability: When the test is repeated 6 timeswith 200 mL of the solution for system suitability test underthe above operating conditions, the relative standard devia-tion of the ratios of the peak area of liothyronine to that ofthe internal standard is not more than 1.0z.

Add the following:

Lysozyme Hydrochloride塩化リゾチーム

C616H963N193O182S10.xHCl[12650-88-3, egg white lysozyme]

Lysozyme Hydrochloride is a hydrochloride of abasic polypeptide obtained from albumen of hen'segg, and has an activity to hydrolyze mucopolysaccha-rides.

It contains not less than 0.9 mg (potency) of lyso-zyme per mg, calculated on the dried basis.

Description Lysozyme Hydrochloride occurs as white,crystals, or crystalline or amorphous powder.


It is hygroscopic.The pH of a solution of Lysozyme Hydrochloride (3 in

200) is between 3.0 and 5.0.

Identiˆcation (1) To 5 mL of a solution of LysozymeHydrochloride in acetate buŠer solution, pH 5.4 (1 in 500)add 1 mL of ninhydrin TS, and heat for 10 minutes: a blue-purple color develops.

(2) Determine the absorption spectrum of a solution ofLysozyme Hydrochloride in acetate buŠer solution, pH 5.4(1 in 10,000) as directed under the Ultraviolet-visible Spec-trophotometry, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of ab-sorption at the same wavelengths.

Purity (1) Clarity of solution—To 5 mL of a solution ofLysozyme Hydrochloride (3 in 200) add, if necessary, dilutehydrochloric acid to adjust the pH to 3: the solution is clear.

(2) Heavy metals—Proceed with 1.0 g of LysozymeHydrochloride according to Method 2, and perform the test.Prepare the control solution with 2.0 mL of Standard LeadSolution (not more than 20 ppm).

Loss on drying Not more than 8.0z (0.1 g, 1059C, 2hours).


Nitrogen Perform the test as directed under the NitrogenDetermination: the amount of nitrogen (N: 14.01) is between16.8z and 18.6z, calculated on the dried basis.

Assay Weigh accurately an amount of LysozymeHydrochloride, equivalent to about 25 mg (potency), dis-solve in phosphate buŠer solution, pH 6.2 to make exactly


100 mL. Pipet 2 mL of this solution, add phosphate buŠersolution, pH 6.2 to make exactly 50 mL, and use this solu-tion as the sample solution. Separately, weigh accurately anamount of Lysozyme Reference Standard (separately deter-mine its loss on drying in the same manner as LysozymeHydrochloride), equivalent to about 25 mg (potency), anddissolve in phosphate buŠer solution, pH 6.2 to make ex-actly 100 mL. Pipet 1 mL and 2 mL of this solution, addphosphate buŠer solution, pH 6.2 to them to make exactly50 mL, and use these solutions as the standard solution (1)and the solution (2), respectively. Keep the sample solutionand the standard solutions in an ice-bath. Pipet 4 mL of sub-strate solution for lysozyme hydrochloride, previouslywarmed in a water bath of 359C for about 5 minutes, add ex-actly 100 mL of the sample solution, previously warmed in awater bath of 359C for about 3 minutes, and allow to standat 359C for exactly 10 minutes, then add exactly 0.5 mL of 1mol/L hydrochloric acid TS, and immediately shake. Deter-mine the absorbance, AT, of this solution at 640 nm, usingwater as the blank. Determine the absorbances, AS1 and AS2,of the solutions obtained with the standard solution (1) andthe standard solution (2) in the same manner as the samplesolution.

Amount [mg (potency)] of lysozyme per mg,calculated on the dried basis

＝WS

2WT×ÅAS1－ AT

AS1－ AS2＋ 1œ

WS: Amount (mg) of Lysozyme Reference Standard, cal-culated on the dried basis.

WT: Amount (mg) of the sample, calculated on the driedbasis.


Magnesium Sulfate Injection硫酸マグネシウム注射液


Description Magnesium Sulfate Injection is a clear, color-less liquid.


pH 5.5 – 7.0 When the labeled concentration exceeds5z, prepare a solution of 5z with water, and perform thetest.

Change the Bacterial endotoxins to read:

Bacterial endotoxins Less than 0.09 EU/mg.



D-Mannitol Injectionマンニトール注射液

Delete the Residue on ignition.

Delete the Pyrogen and add the following:

Bacterial endotoxins Less than 0.50 EU/mL.



Menatetrenoneメナテトレノン


Description Menatetrenone occurs as yellow, crystals,crystalline powder, waxy mass or oily material.

It is very soluble in hexane, soluble in ethanol (99.5), spar-ingly soluble in 2-propanol, slightly soluble in methanol, andpractically insoluble in water.

It decomposes and the color becomes more intense bylight.


Mepitiostaneメピチオスタン


Assay Weigh accurately about 0.3 g of Mepitiostane, anddissolve in cyclohexane to make exactly 10 mL. Pipet 2 mLof this solution, add 10 mL of ethanol (99.5), mix with ex-actly 2 mL each of 0.01 molWL hydrochloric acid TS and theinternal standard solution, add ethanol (99.5) to make 20mL, allow to stand at ordinary temperature for 30 minutes,and use this solution as the sample solution. Separately,weigh accurately about 45 mg of Epitiostanol ReferenceStandard, dissolve in exactly 2 mL of the internal standardsolution, add ethanol (99.5) to make 20 mL, and use this so-lution as the standard solution. Perform the test with 10 mLeach of the sample solution and the standard solution asdirected under the Liquid Chromatography according to thefollowing conditions, and calculate the ratios, QT and QS, ofthe peak area of epitiostanol to that of the internal standard,respectively.

Amount (mg) of C25H40O2S＝WS×QT

QS× 5× 1.3202

WS: Amount (mg) of Epitiostanol Reference Standard,calculated on the anhydrous basis

Internal standard solution—A solution of n-octylbenzene in


ethanol (99.5) (1 in 300).Operating conditions—





of epitiostanol is about 6 minutes.System suitability—

System performance: When the procedure is run with10 mL of the standard solution under the above operatingconditions, epitiostanol and the internal standard are elutedin this order with the resolution between these peaks beingnot less than 4.

System repeatability: When the test is repeated 6 timeswith 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratiosof the peak area of epitiostanol to that of the internal stan-dard is not more than 1.0z.

Methotrexateメトトレキサート


Assay Weigh accurately about 25 mg each of Methotrexateand Methotrexate Reference Standard, dissolve in the mo-bile phase to make exactly 250 mL, and use these solutionsas the sample solution and the standard solution. Performthe test with 10 mL each of these solutions as directed underthe Liquid Chromatography according to the following con-ditions, and measure the peak areas, AT and AS, of meth-otrexate in each solution.

Amount (mg) of C20H22N8O5＝WS×AT

AS

WS: Amount (mg) of Methotrexate Reference Standard,calculated on the anhydrous basis





Mobile phase: A mixture of disodium hydrogen phos-phate-citric acid buŠer solution, pH 6.0 and acetonitrile(89:11).

Flow rate: Adjust the ‰ow rate so that the retention time

of methotrexate is about 8 minutes.System suitability—

System performance: Dissolve 10 mg each of Meth-otrexate and folic acid in 100 mL of the mobile phase. Whenthe procedure is run with 10 mL of this solution under theabove operating conditions, folic acid and methotrexate areeluted in this order with the resolution between these peaksbeing not less than 8.

System repeatability: When the test is repeated 6 timeswith 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peakarea of methotrexate is not more than 1.0z.

Meticraneメチクラン


Purity(4) Related substances—Dissolve 0.05 g of Meticrane in

50 mL of acetonitrile, and use this solution as the sample so-lution. Pipet 1 mL of the sample solution, add acetonitrile tomake exactly 100 mL, and use this solution as the standardsolution. Perform the test with exactly 2 mL each of the sam-ple solution and the standard solution as directed under theLiquid Chromatography according to the following condi-tions, and determine each peak area of both solutions by theautomatic integration method: the total area of the peaksother than meticrane from the sample solution is not largerthan the peak area of meticrane from the standard solution.Operating conditions 1—





of meticrane is about 7 minutes.Time span of measurement: About 4 times as long as the

retention time of meticrane, after the solvent peak.System suitability 1—

Test for required detection: To exactly 2 mL of the stan-dard solution add the mobile phase to make exactly 20 mL.Conˆrm that the peak area of meticrane obtained from10 mL of this solution is equivalent to 7 to 13z of that ofmeticrane obtained from 10 mL of the standard solution.

System performance: Dissolve 0.01 g each of Meticraneand caŠeine in 100 mL of acetonitrile. To exactly 2 mL ofthis solution add the mobile phase to make exactly 10 mL.When the procedure is run with 10 mL of this solution underthe above operating conditions 1, caŠeine and meticrane areeluted in this order with the resolution between these peaks


being not less than 10.System repeatability: When the test is repeated 6 times

with 10 mL of the standard solution under the above operat-ing conditions 1, the relative standard deviation of the peakarea of meticrane is not more than 2.0z.Operating conditions 2—

Detector, column, and column temperature: Proceed asdirected in the operating conditions 1.


of meticrane is about 2 minutes.Time span of measurement: About 10 times as long as the

retention time of meticrane, after the solvent peak.System suitability 2—

Test for required detection: To exactly 2 mL of the stan-dard solution add the mobile phase to make exactly 20 mL.Conˆrm that the peak area of meticrane obtained from10 mL of this solution is equivalent to 7 to 13z of that ofmeticrane obtained from 10 mL of the standard solution.

System performance: Dissolve 0.02 g each of Meticraneand methyl parahydroxybenzoate in 100 mL of acetonitrile.To exactly 2 mL of this solution add the mobile phase tomake exactly 10 mL. When the procedure is run with 10 mLof this solution under the above operating conditions 2,meticrane and methyl parahydroxybenzoate are eluted inthis order with the resolution between these peaks being notless than 4.

System repeatability: When the test is repeated 6 timeswith 10 mL of the standard solution under the above operat-ing conditions 2, the relative standard deviation of the peakarea of meticrane is not more than 2.0z.

Micronomicin Sulfate硫酸ミクロノマイシン


Micronomicin Sulfate contains not less than 590 mg(potency) per mg, calculated on the anhydrous basis.The potency of Micronomicin Sulfate is expressed asmass (potency) of micronomicin (C20H41N5O7:463.57).

Description Micronomicin Sulfate occurs as a white tolight yellowish white powder.

It is very soluble in water, sparingly soluble in ethyleneglycol, and practically insoluble in methanol and in ethanol(99.5).

It is hygroscopic.

Identiˆcation (1) Dissolve 50 mg each of MicronomicinSulfate and Micronomicin Sulfate Reference Standard in 10mL of water, and use these solutions as the sample solutionand the standard solution. Perform the test with these solu-tions as directed under the Thin-layer Chromatography.Spot 5 mL of the sample solution and the standard solution

on a plate of silica gel for thin-layer chromatography. De-velop the plate with a mixture of ethanol (99.5), 1-buthanoland ammonia solution (28) (10:8:7) to a distance of about 10cm, and air-dry the plate. Spray evenly a solution of nin-hydrin in a mixture of acetone and pyridine (25:1) (1 in 500),and heat at 1009C for 10 minutes: the spots obtained fromthe sample solution and the standard solution are red-purpleto red-brown and their Rf values are the same.

(2) To 5 mL of a solution of Micronomicin Sulfate (1 in100) add 1 mL of barium chloride TS: a white precipitate isformed, and it does not dissolve by addition of dilute nitricacid.

Optical rotation [a]20D : ＋110 –＋1309(0.25 g calculated

on the anhydrous basis, water, 25 mL, 100 mm).

pH The pH of a solution obtained by dissolving 1.0 g ofMicronomicin Sulfate in 10 mL of water is between 3.5 and5.5.

Purity (1) Clarity and color of solution—Dissolve 1.5 gof Micronomicin Sulfate in 10 mL of water: the solution isclear and colorless to pale yellow.

(2) Heavy metals—Proceed with 1.0 g of MicronomicinSulfate according to Method 2, and perform the test. Pre-pare the control solution with 2.0 mL of Standard Lead So-lution (not more than 20 ppm).

(3) Related substances—Dissolve 0.40 g of Micronomi-cin Sulfate in 10 mL of water, and use this solution as thesample solution. Pipet 1 mL of the sample solution, addwater to make exactly 200 mL, and use this solution as thestandard solution. Perform the test with these solutions asdirected under the Thin-layer Chromatography. Spot 5 mLof the sample solution and the standard solution on a plateof silica gel for thin-layer chromatography. Develop theplate with a mixture of ethanol (99.5), 1-buthanol and am-monia solution (28) (10:8:7) to a distance of about 10 cm,and air-dry the plate. Spray evenly a solution of ninhydrin ina mixture of acetone and pyridine (25:1) (1 in 500), and heatat 1009C for 10 minutes: the spot other than the principalspot obtained from the sample solution is not more intensethan the spot from the standard solution.

Water Not more than 10.0z (0.2 g, volumetric titration,back titration). Use a mixture of methanol for water deter-mination and ethylene glycol for water determination (1:1)instead of methanol for water determination.




(3) Standard solutions—Weigh accurately an amount ofMicronomicin Sulfate Reference Standard, equivalent toabout 20 mg (potency), dissolve in 0.1 mol/L phosphatebuŠer solution for antibiotics, pH 8.0 to make exactly 20mL, and use this solution as the standard stock solution.


Keep the standard stock solution at 5 – 159C, and use within30 days. Take exactly a suitable amount of the standardstock solution before use, add 0.1 mol/L phosphate buŠersolution for antibiotics, pH 8.0 to make solutions so thateach mL contains 2 mg (potency) and 0.5 mg (potency), anduse these solutions as the high concentration standard solu-tion and the low concentration standard solution, respec-tively.

(4) Sample solutions—Weigh accurately an amount ofMicronomicin Sulfate, equivalent to about 20 mg (potency),and dissolve in 0.1 mol/L phosphate buŠer solution for an-tibiotics, pH 8.0 to make exactly 20 mL. Take exactly a suit-able amount of this solution, add 0.1 mol/L phosphatebuŠer solution for antibiotics, pH 8.0 to make solutions sothat each mL contains 2 mg (potency) and 0.5 mg (potency),and use these solutions as the high concentration sample so-lution and the low concentration sample solution, respec-tively.


Migreninミグレニン


Assay(2) CaŠeine—To about 1 g of Migrenin, previously

dried and accurately weighed, add exactly 5 mL of the inter-nal standard solution, dissolve in chloroform to make 10mL, and use this solution as the sample solution. Separately,weigh accurately about 90 mg of CaŠeine Reference Stan-dard, previously dried at 809C for 4 hours, add exactly 5 mLof the internal standard solution, dissolve in chloroform tomake 10 mL, and use this solution as the standard solution.Perform the test with 1 mL each of the sample solution andthe standard solution as directed under the Gas Chromatog-raphy according to the following conditions, and calculatethe ratios, QT and QS, of the peak area of caŠeine to that ofthe internal standard.

Amount (mg) of caŠeine (C8H10N4O2)＝WS×QT

QS

WS: Amount (mg) of CaŠeine Reference Standard

Internal standard solution—A solution of ethenzamide inchloroform (1 in 50).Operating conditions—


210 cm in length, packed with siliceous earth for gas chro-matography (180 to 250 mm in particle diameter) coated with50z phenyl-methyl silicon polymer for gas chromatographyat the ratio of 15z.


Carrier gas: Nitrogen

Flow rate: Adjust the ‰ow rate so that the retention timeof ethenzamide is about 4 minutes.System suitability—

System performance: Dissolve 0.9 g of antipyrine and0.09 g of caŠeine in 10 mL of chloroform. When the proce-dure is run with 1 mL of this solution under the above operat-ing conditions, caŠeine and antipyrine are eluted in this ord-er with the resolution between these peaks being not lessthan 1.5.

System repeatability: When the test is repeated 6 timeswith 1 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratiosof the peak area of caŠeine to that of the internal standard isnot more than 1.0z.

Minocycline Hydrochloride塩酸ミノサイクリン


Identiˆcation (1) Determine the absorption spectrum ofa solution of Minocycline Hydrochloride in a solution ofhydrochloric acid in methanol (19 in 20,000) (1 in 62,500) asdirected under the Ultraviolet-visible Spectrophotometry,and compare the spectrum with the Reference Spectrum orthe spectrum of a solution of Minocycline HydrochlorideReference Standard prepared in the same manner as thesample solution: both spectra exhibit similar intensities ofabsorption at the same wavelengths.

(2) Determine the infrared absorption spectrum ofMinocycline Hydrochloride as directed in the potassiumchloride disk method under the Infrared Spectrophotomet-ry, and compare the spectrum with the Reference Spectrumor the spectrum of Minocycline Hydrochloride ReferenceStandard: both spectra exhibit similar intensities of absorp-tion at the same wave numbers.

(3) A solution of Minocycline Hydrochloride (1 in 100)responds to the Qualitative Test (2) for chloride.

Delete the Absorbance.


Purity (1) A solution of Minocycline Hydrochloride (1 in100) is clear, and when the test is performed within 1 hourafter preparation of this solution, the absorbance of the so-lution at 560 nm, determined as directed under the Ultrav-iolet-visible Spectrophotometry, is not more than 0.06.

(2) Heavy metals—Proceed with 0.5 g of MinocyclineHydrochloride according to Method 2, and perform the test.Prepare the control solution with 2.5 mL of Standard LeadSolution (not more than 50 ppm).

(3) Related substances—Dissolve 50 mg of MinocyclineHydrochloride in 100 mL of the mobile phase, and use thissolution as the sample solution. Perform the test immedi-ately after the preparation of the sample solution with 20 mLof the sample solution as directed under the Liquid Chro-


matography according to the following conditions, andmeasure each peak area by the automatic integrationmethod. Calculate the amount of each peak area by the areapercentage method: the amount of epiminocycline is notmore than 1.2z, the amount of each peak other thanminocycline and epiminocycline is not more than 1.0z, andthe total area of the peaks other than minocycline andepiminocycline is not more than 2.0z.Operating conditions—

Detector, column, column temperature, and mobilephase: Proceed as directed in the operating conditions in theAssay.

Flow rate: Adjust the ‰ow rate so that the retention timeof minocycline is about 12 minutes. The retention time ofepiminocycline is about 10 minutes under this condition.

Time span of measurement: About 2.5 times as long as theretention time of minocycline after the solvent peak.System suitability—

Test for required detection: To exactly 2 mL of the samplesolution add the mobile phase to make exactly 100 mL, anduse this solution as the solution for system suitability test.Pipet 5 mL of the solution for system suitability test, andadd the mobile phase to make exactly 100 mL. Conˆrm thatthe peak area of minocycline obtained from 20 mL of this so-lution is equivalent to 3.5 to 6.5z of that of minocycline ob-tained from 20 mL of the solution for system suitability test.


System repeatability: When the test is repeated 6 timeswith 20 mL of the solution for system suitability test underthe above operating conditions, the relative standard devia-tion of the peak area of minocycline is not more than 2.0z.

Mitomycin CマイトマイシンC


Mitomycin C contains not less than 970 mg (potency)per mg, calculated on the dried basis. The potency ofMitomycin C is expressed as mass (potency) ofmitomycin C (C15H18N4O5).

Description Mitomycin C occurs as blue-purple, crystals orcrystalline powder.

It is freely soluble in N,N-dimethylacetamide, slightlysoluble in water and in methanol, and very slightly soluble inethanol (99.5).

Identiˆcation (1) Determine the absorption spectrum ofa solution of Mitomycin C (1 in 100,000) as directed underthe Ultraviolet-visible Spectrophotometry, and compare thespectrum with the Reference Spectrum or the spectrum of asolution of Mitomycin C Reference Standard prepared in thesame manner as the sample solution: both spectra exhibitsimilar intensities of absorption at the same wavelengths.

(2) Determine the infrared absorption spectrum ofMitomycin C as directed in the potassium bromide diskmethod under the Infrared Spectrophotometry, and com-pare the spectrum with the Reference Spectrum or the spec-trum of Mitomycin C Reference Standard: both spectraexhibit similar intensities of absorption at the same wavenumbers.

Purity Related substances—Conduct this procedure rap-idly after the sample and the standard solutions are pre-pared. Dissolve 50 mg of Mitomycin C in 10 mL ofmethanol, and use this solution as the sample solution. Pipet1 mL of the sample solution, add methanol to make exactly100 mL, and use this solution as the standard solution. Per-form the test with exactly 10 mL each of the sample solutionand the standard solution as directed under the Liquid Chro-matography according to the following conditions, and de-termine each peak area by the automatic integrationmethod: each area of the peak other than mitomycin C ob-tained from the sample solution is not more than the peakarea of mitomycin C from the standard solution, and thetotal area of the peaks other than mitomycin C from thesample solution is not more than 3 times the peak area ofmitomycin C from the standard solution.Operating conditions—




Mobile phase A: To 20 mL of 0.5 mol/L ammoniumacetate TS add water to make 1000 mL. To 800 mL of thissolution add 200 mL of methanol.

Mobile phase B: To 20 mL of 0.5 mol/L ammoniumacetate TS add water to make 1000 mL. To this solution add1000 mL of methanol.

Flowing of the mobile phase: Control the gradient by mix-ing the mobile phases A and B as directed in the following t-able.


Mobile phaseA (z)

Mobile phaseB (z)

0 – 10 100 010 – 30 100→ 0 0→ 10030 – 45 0 100

Flow rate: About 1.0 mL/minTime span of measurement: About 2 times as long as the

retention time of mitomycin C after the solvent peak.System suitability—

Test for required detection: Pipet 10 mL of the standardsolution, and add methanol to make exactly 100 mL. Con-ˆrm that the peak area of mitomycin C obtained from 10 mLof this solution is equivalent to 7 to 13z of that from 10 mLof the standard solution.

System performance: Dissolve 25 mg of Mitomycin C and40 mg of 3-ethoxy-4-hydroxybenzaldehyde in 50 mL of


methanol. When the procedure is run with 10 mL of this so-lution under the above operating conditions, mitomycin Cand 3-ethoxy-4-hydroxybenzaldehyde are eluted in this orderwith the resolution between these peaks being not less than15.

System repeatability: When the test is repeated 3 timeswith 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peakarea of mitomycin C is not more than 3.0z.


Assay Weigh accurately an amount of Mitomycin C andMitomycin C Reference Standard, equivalent to about25 mg (potency), dissolve each in N,N-dimethylacetamide tomake exactly 50 mL, and use these solutions as the samplesolution and the standard solution. Perform the test with ex-actly 10 mL each of the sample solution and the standard so-lution as directed under the Liquid Chromatography accord-ing to the following conditions, and determine the peakareas, AT and AS, of mitomycin C.


＝WS×AT

AS× 1000

WS: Amount [mg (potency)] of Mitomycin C ReferenceStandard



and 30 cm in length, packed with phenylated silica gel forliquid chromatography (10 mm in particle diameter).


Mobile phase: To 40 mL of 0.5 mol/L ammonium acetateTS add 5 mL of diluted acetic acid (100) (1 in 20) and waterto make 1000 mL. To 600 mL of this solution add 200 mL ofmethanol.

Flow rate: Adjust the ‰ow rate so that the retention timeof mitomycin C is about 7 minutes.System suitability—

System performance: Dissolve about 25 mg of MitomycinC Reference Standard and about 0.375 g of 3-ethoxy-4-hydroxybenzaldehyde in 50 mL of N,N-dimethylacetamide.When the procedure is run with 10 mL of this solution underthe above operating conditions, mitomycin C and 3-ethoxy-4-hydroxybenzaldehyde are eluted in this order with theresolution between these peaks being not less than 3.

System repeatability: When the test is repeated 6 timeswith 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peakarea of mitomycin C is not more than 1.0z.


Morphine Hydrochloride塩酸モルヒネ


Description Morphine Hydrochloride occurs as white,crystals or crystalline powder.

It is freely soluble in formic acid, soluble in water, spar-ingly soluble in methanol, and slightly soluble in ethanol(95).

It is colored by light.

Add the following next to Optical rotation:

pH The pH of a solution obtained by dissolving 0.10 g ofMorphine Hydrochloride in 10 mL of water is between 4.0and 6.0.


Purity (1) Clarity and color of solution—Dissolve 0.10 gof Morphine Hydrochloride in 10 mL of water: the solutionis clear and colorless.

(2) Sulfate—Dissolve 0.20 g of Morphine Hydrochloridein 5 mL of water, and add 2 to 3 drops of barium chlorideTS: no turbidity is produced.

(3) Meconic acid—Dissolve 0.20 g of MorphineHydrochloride in 5 mL of water, and add 5 mL of dilutehydrochloric acid and 2 drops of iron (III) chloride TS: nored color develops.

(4) Other alkaloids—Dissolve 0.1 g of MorphineHydrochloride in 10 mL of diluted ethanol (95) (1 in 2), anduse this solution as the sample solution. Pipet 1 mL of thesample solution, add diluted ethanol (95) (1 in 2) to make ex-actly 200 mL, and use this solution as the standard solution.Perform the test with these solutions as directed under theThin-layer Chromatography. Spot 10 mL each of the samplesolution and the standard solution on a plate of silica gelwith ‰uorescent indicator for thin-layer chromatography.Develop the plate with a mixture of ethanol (99.5), toluene,acetone and ammonia solution (28) (14:14:7:1) to a distanceof about 15 cm, and air-dry the plate. Examine under ultrav-iolet light (main wavelength: 254 nm): the spots other thanthe principal spot from the sample solution are not more in-tense than the spot from the standard solution.

Morphine Hydrochloride Injection塩酸モルヒネ注射液


Assay Take exactly a volume of Morphine HydrochlorideInjection, equivalent to about 80 mg of morphinehydrochloride (C17H19NO3.HCl.3H2O), and add water tomake exactly 20 mL. Pipet 5 mL of this solution, add exactly10 mL of the internal standard solution and water to make50 mL, and use this solution as the sample solution.


Separately, weigh accurately about 25 mg of morphinehydrochloride for assay, dissolve in exactly 10 mL of the in-ternal standard solution, add water to make 50 mL, and usethis solution as the standard solution. Perform the test with20 mL each of the sample solution and the standard solutionas directed under the Liquid Chromatography according tothe following conditions, and calculate the ratios, QT andQS, of the peak area of morphine to that of the internal stan-dard.

Amount (mg) of morphine hydrochloride(C17H19NO3.HCl.3H2O)

＝WS×QT

QS× 4× 1.1679

WS: Amount (mg) of morphine hydrochloride for assay,calculated on the anhydrous basis

Internal standard solution—A solution of etilefrinehydrochloride (1 in 500).Operating conditions—





Flow rate: Adjust the ‰ow rate so that retention time ofmorphine is about 10 minutes.System suitability—

System performance: When the procedure is run with20 mL of the standard solution under the above operatingconditions, morphine and the internal standard are eluted inthis order with the resolution between these peaks being notless than 3.

System repeatability: When the test is repeated 6 timeswith 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratiosof the peak area of morphine to that of the internal standardis not more than 1.0z.

Morphine Hydrochloride Tablets塩酸モルヒネ錠


Assay Take not less than 20 Morphine HydrochlorideTablets, weigh accurately, and powder. Weigh accurately aquantity of the powder, equivalent to about 20 mg of mor-phine hydrochloride (C17H19NO3.HCl.3H2O), add exactly10 mL of the internal standard solution, extract the mixturewith ultrasonic waves for 10 minutes, and add water to make

50 mL. Filter this solution, and use the ˆltrate as the samplesolution. Separately, weigh accurately about 25 mg of mor-phine hydrochloride for assay, dissolve in exactly 10 mL ofthe internal standard solution, add water to make 50 mL,and use this solution as the standard solution. Perform thetest with 20 mL each of the sample solution and the standardsolution as directed under the Liquid Chromatography ac-cording to the following conditions, and calculate the ratios,QT and QS, of the peak area of morphine to that of the inter-nal standard.


＝WS×QT

QS× 1.1679


Internal standard solution—A solution of etilefrinehydrochloride (1 in 500).Operating conditions—





Flow rate: Adjust the ‰ow rate so that the retention timeof morphine is about 10 minutes.System suitability—

System performance: When the procedure is run with20 mL of the standard solution under the above operatingconditions, morphine and the internal standard are eluted inthis order with the resolution between these peaks being notless than 3.


Norepinephrineノルエピネフリン


Description Norepinephrine occurs as a white to lightbrown or slightly reddish brown, crystalline powder.

It is freely soluble in acetic acid (100), very slightly solublein water, and practically insoluble in ethanol (95).

It dissolves in dilute hydrochloric acid.


It gradually changes to brown by air and by light.


Identiˆcation (1) Determine the absorption spectrum ofa solution of Norepinephrine in 0.1 mol/L hydrochloric acidTS (3 in 100,000) as directed under the Ultraviolet-visibleSpectrophotometry, and compare the spectrum with theReference Spectrum: both spectra exhibit similar intensitiesof absorption at the same wavelengths.

(2) Determine the infrared absorption spectrum ofNorepinephrine, previously dried, as directed in the potassi-um bromide disk method under the Infrared Spectrophoto-metry, and compare the spectrum with the Reference Spec-trum: both spectra exhibit similar intensities of absorption atthe same wave numbers.

Norepinephrine Injectionノルエピネフリン注射液


Identiˆcation Transfer a volume of Norepinephrine Injec-tion, equivalent to 1 mg of Norepinephrine according to thelabeled amount, to each of two test tubes A and B, and add1 mL of water to each tube. Add 10 mL of potassium hydro-gen phthalate buŠer solution, pH 3.5, to A, and 10 mL ofphosphate buŠer solution, pH 6.5, to B. To each of these so-lutions add 1.0 mL of iodine TS, allow to stand for 5minutes, and add 2.0 mL of sodium thiosulfate TS: no coloror a pale red color develops in test tube A, and a deep red-purple color develops in test tube B.

Norgestrel and EthinylestradiolTabletsノルゲストレルエチニルエストラジオール錠

Change the Content uniformity to read:

Content uniformity Add 2 mL of diluted methanol (7 in10) to 1 tablet of Norgestrel and Ethinylestradiol Tablets,add exactly 2 mL of the internal standard solution, shake for20 minutes, and centrifuge. Filter the supernatant liquidthrough a membrane ˆlter with pore size of not more than0.2 mm, and use this ˆltrate as the sample solution. Separate-ly, weigh accurately quantities of Norgestrel Reference Stan-dard and of Ethinylestradiol Reference Standard, equivalentto 100 times each of the labeled amounts, dissolve in dilutedmethanol (7 in 10) to make exactly 200 mL. Pipet 2 mL ofthis solution, add exactly 2 mL of the internal standard solu-tion, and use this solution as the standard solution. Performthe test with 20 mL each of the sample solution and the stan-dard solution as directed under the Liquid Chromatographyaccording to the following conditions. Calculate the ratios,QTa and QTb, of the peak areas of norgestrel and ethinyles-

tradiol to the peak area of the internal standard of the sam-ple solution and also the ratios, QSa and QSb, of the peakareas of norgestrel and ethinylestradiol to the peak area ofthe internal standard of the standard solution.

Amount (mg) of norgestrel (C21H28O2)

＝WSa×QTa

QSa×

1100

Amount (mg) of ethinylestradiol (C20H24O2)

＝WSb×QTb

QSb×

1100

WSa: Amount (mg) of Norgestrel Reference StandardWSb: Amount (mg) of Ethinylestradiol Reference

Standard

Internal standard solution—A solution of diphenyl in dilut-ed methanol (7 in 10) (1 in 50,000).Operating conditions—

Proceed as directed in the operating conditions in theAssay.System suitability—


Change the Dissolution test to read:

Dissolution test Perform the test with 1 tablet of Norges-trel and Ethinylestradiol Tablets at 50 revolutions perminute according to Method 2 under the Dissolution Test,using 900 mL of water as the test solution. Take 50 mL ormore of the dissolved solution 45 minutes after starting thetest, and membrane ˆlter through a membrane ˆlter withpore size of not more than 0.8 mm. Discard the ˆrst 10 mL ofthe ˆltrate, transfer exactly 30 mL of the subsequent into achromatography column [prepared by packing 0.36 g of oc-tadecylsilanized silica gel for pretreatment (55 to 105 mm inparticle diameter) in a tube about 1 cm in inside diameter].After washing the column with 15 mL of water, elute with3 mL of methanol, and evaporate the eŒuent on a waterbath to dryness at about 409C with the aid of a current air.Dissolve the residue in exactly 2 mL of diluted methanol (7in 10), and use this solution as the sample solution. Separate-ly, weigh accurately about 25 mg of Norgestrel ReferenceStandard and about 2.5 mg of Ethinylestradiol ReferenceStandard dissolve in diluted methanol (7 in 10) to make ex-actly 100 mL, then pipet 3 mL of this solution, add dilutedmethanol (7 in 10) to make exactly 100 mL, and use this so-lution as the standard solution. Perform the test with exactly50 mL each of the sample solution and the standard solutionas directed under the Liquid Chromatography according tothe following conditions. Determine the peak areas, ATa andATb, of norgestrel and ethinylestradiol from the sample solu-tion, and the peak areas, ASa and ASb, of norgestrel andethinylestradiol from the standard solution.

The dissolution rate of Norgestrel and EthinylestradiolTablets in 45 minutes is not less than 70z.


Dissolution rate (z) with respect to the labeled amountof norgestrel (C21H28O2)

＝WSa×ATa

ASa×

1Ca×

95

Dissolution rate (z) with respect to the labeled amountof ethinylestradiol (C20H24O2)

＝WSb×ATb

ASb×

1Cb×

95

WSa: Amount (mg) of Norgestrel Reference StandardWSb: Amount (mg) of Ethinylestradiol Reference Stand-

ardCa: Labeled amount (mg) of norgestrel (C21H28O2) in 1

tabletCb: Labeled amount (mg) of ethinylestradiol (C20H24O2)

in 1 tablet

Operating conditions—Proceed as directed in the operating conditions in the As-

say.System suitability—



Assay Weigh accurately not less than 20 Norgestrel andEthinylestradiol Tablets, and powder. Weigh accurately aportion of the powder, equivalent to about 1 mg of norges-trel (C21H28O2), add 4 mL of diluted methanol (7 in 10), addexactly 4 mL of the internal standard solution, shake for 20minutes, and centrifuge. Filter the supernatant liquidthrough a membrane ˆlter with pore size of not more than0.2 mm, and use this ˆltrate as the sample solution. Separate-ly, weigh accurately about 50 mg of Norgestrel ReferenceStandard and about 5 mg of Ethinylestradiol ReferenceStandard, and dissolve in diluted methanol (7 in 10) to makeexactly 200 mL. Pipet 4 mL of this solution, add exactly 4mL of the internal standard solution, and use this solution asthe standard solution. Perform the test with 20 mL each ofthe sample solution and the standard solution as directed un-der the Liquid Chromatography according to the followingconditions. Calculate the ratios, QTa and QTb, of the peakareas of norgestrel and ethinylestradiol to the peak area ofthe internal standard of the sample solution and also the ra-tios, QSa and QSb, of the peak areas of norgestrel andethinylestradiol to the peak area of the internal standard ofthe standard solution.

Amount (mg) of norgestrel (C21H28O2)

＝WSa×QTa

QSa×

150

Amount (mg) of ethinylestradiol (C20H24O2)

＝WSb×QTb

QSb×

150

WSa: Amount (mg) of Norgestrel Reference StandardWSb: Amount (mg) of Ethinylestradiol Reference Stand-

ard

Internal standard solution—A solution of diphenyl in dilut-ed methanol (7 in 10) (1 in 50,000).

Operating conditions—Detector: Norgestrel—An ultraviolet absorption photom-

eter (wavelength: 241 nm).Ethinylestradiol—A ‰uorophotometer (excitation wave-

length: 281 nm, ‰uorescence wavelength: 305 nm).Column: A stainless steel column 4.6 mm in inside di-




of norgestrel is about 10 minutes.System suitability—

System performance: When the procedure is run with 20mL of the standard solution under the above operating con-ditions, ethinylestradiol, norgestrel and the internal stan-dard are eluted in this order, and the resolution between thepeaks of norgestrel and the internal standard is not less than8.

System repeatability: When the test is repeated 6 timeswith 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratiosof the peak area of ethinylestradiol and norgestrel to that ofthe internal standard are not more than 1.0z, respectively.

Add the following:

O‰oxacinオフロキサシン

C18H20FN3O4: 361.37(3RS )-9-Fluoro-2,3-dihydro-3-methyl-10-(4-methylpiperazin-1-yl)-7-oxo-7H-pyrido[1,2,3-de]-1,4-benzoxazine-6-carboxylic acid [82419-36-1 ]

O‰oxacin, when dried, contains not less than99.0z and not more than 101.0z of o‰oxacin(C18H20FN3O4).

Description O‰oxacin occurs as pale yellowish white tolight yellowish white, crystals or crystalline powder.

It is freely soluble in acetic acid (100), slightly soluble inwater, and very slightly soluble in acetonitrile and in ethanol(99.5).

A soluton of O‰oxacin in sodium hydroxide TS (1 in 20)does not show optical rotation.

It is changed in color by light.Melting point: about 2659C (with decomposition).

Identiˆcation (1) Determine the absorption spectrum of


a solution of O‰oxacin in 0.1 mol/L hydrochloric acid TS (1in 150,000) as directed under the Ultraviolet-visible Spec-trophotometry, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of ab-sorption at the same wavelengths.

(2) Determine the infrared absorption spectrum ofO‰oxacin as directed in the potassium bromide disk methodunder the Infrared Spectrophotometry, and compare thespectrum with the Reference Spectrum: both spectra exhibitsimilar intensities of absorption at the same wave numbers.

Purity (1) Heavy metals—Proceed with 2.0 g of O‰oxa-cin according to Method 4, and perform the test. Prepare thecontrol solution with 2.0 mL of Standard Lead Solution (notmore than 10 ppm).

(2) Related substances—Conduct this procedure withoutexposure to light. Dissolve 10 mg of O‰oxacin in 50 mL of amixture of water and acetonitrile (6:1), and use this solutionas the sample solution. Pipet 1 mL of the sample solution,and add a mixture of water and acetonitrile (6:1) to make ex-actly 20 mL. Pipet 1 mL of this solution, add a mixture ofwater and acetonitrile (6:1) to make exactly 10 mL, and usethis solution as the standard solution. Perform the test withexactly 10 mL each of the sample solution and the standardsolution as directed under the Liquid Chromatography ac-cording to the following conditions, and determine eachpeak area by the automatic integration method: the area ofthe peak other than o‰oxacin obtained from the sample so-lution is not more than 0.4 times the peak area of o‰oxacinfrom the standard solution, and the total area of the peaksother than o‰oxacin from the sample solution is not morethan the peak area from the standard solution.Operating conditions—




Mobile phase: Dissolve 7.0 g of sodium perchloratemonohydrate and 4.0 g of ammonium acetate in 1300 mL ofwater, adjust the pH to 2.2 with phosphoric acid, and add240 mL of acetonitrile.

Flow rate: Adjust the ‰ow rate so that the retention timeof o‰oxacin is about 20 minutes.

Time span of measurement: About 1.8 times as long as theretention time of o‰oxacin after the solvent peak.System suitability—

Test for required detectability: Measure 1 mL of the stan-dard solution, and add a mixture of water and acetonitrile(6:1) to make exactly 20 mL. Conˆrm that the peak area ofo‰oxacin obtained from 10 mL of this solution is equivalentto 4 to 6z of that from 10 mL of the standard solution.

System performance: To 0.5 mL of the sample solutionadd 1 mL of a solution of o‰oxacin demethyl substance in amixture of water and acetonitrile (6:1) (1 in 20,000) and a

mixture of water and acetonitrile (6:1) to make 100 mL.When the procedure is run with 10 mL of this solution underthe above operating conditions, o‰oxacin demethyl sub-stance and o‰oxacin are eluted in this order with the resolu-tion between these peaks being not less than 2.5.

System repeatability: When the test is repeated 6 timeswith 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peakarea of o‰oxacin is not more than 2.0z.

Loss on drying Not less than 0.2z (1 g, 1059C, 4 hours).


Assay Weigh accurately about 0.30 g of O‰oxacin, previ-ously dried, dissolve in 100 mL of acetic acid (100), and ti-trate with 0.1 mol/L perchloric acid VS (potentiometrictitration). Perform a blank determination, and make anynecessary correction.

Each mL of 0.1 mol/L perchloric acid VS＝ 36.14 mg of C18H20FN3O4


Oxytetracycline Hydrochloride塩酸オキシテトラサイクリン


Oxytetracycline Hydrochloride contains not lessthan 880 mg (potency) per mg, calculated on the driedbasis. The potency of Oxytetracycline Hydrochlorideis expressed as mass (potency) of oxytetracycline(C22H24N2O9: 460.43).

Description Oxytetracycline Hydrochloride occurs as yel-low, crystals or crystalline powder.


Identiˆcation (1) Determine the absorption spectrum ofa solution of Oxytetracycline Hydrochloride in 0.1 mol/Lhydrochloric acid TS (1 in 50,000) as directed under theUltraviolet-visible Spectrophotometry, and compare thespectrum with the Reference Spectrum or the spectrum ofOxytetracycline Hydrochloride Reference Standard pre-pared in the same manner as the sample solution: both spec-tra exhibit similar intensities of absorption at the samewavelengths.

(2) Dissolve 20 mg of Oxytetracycline Hydrochloride in3 mL of water, and add 1 drop of silver nitrate TS: a whiteturbidity is produced.

Optical rotation [a]D20: －188 –－2009(0.25 g calculatedon the dried basis, 0.1 mol/L hydrochloric acid, 25 mL, 100mm).


Purity (1) Heavy metals—Proceed with 0.5 g of Oxyt-etracycline Hydrochloride according to Method 2, and per-form the test. Prepare the control solution with 2.5 mL ofStandard Lead Solution (not more than 50 ppm).

(2) Related substances—Dissolve 20 mg of Oxytetracy-cline Hydrochloride in 0.01 mol/L hydrochloric acid TS tomake exactly 25 mL, and use this solution as the sample so-lution. Separately, dissolve 20 mg of 4-epioxytetracycline in0.01 mol/L hydrochloric acid TS to make exactly 25 mL,and use this solution as 4-epioxytetracycline stock solution.Separately, dissolve 20 mg of tetracycline hydrochloride in0.01 mol/L hydrochloric acid TS to make exactly 25 mL,and use this solution as tetracycline hydrochloride stock so-lution. Separately, dissolve 8 mg of b-apooxytetracycline in5 mL of 0.01 mol/L sodium hydroxide TS, add 0.01 mol/Lhydrochloric acid TS to make exactly 100 mL, and use thissolution as b-apooxytetracycline stock solution. Pipet 1 mLof 4-epioxytetracycline stock solution, 4 mL of tetracyclinehydrochloride stock solution and 40 mL of b-apooxytetracy-cline stock solution, add 0.01 mol/L hydrochloric acid TS tomake exactly 200 mL, and use this solution as the standardsolution. Perform the test with exactly 20 mL each of thesample solution and the standard solution as directed underthe Liquid Chromatography according to the following con-ditions, and determine each peak area by the automatic in-tegration method: the peak areas of 4-epioxytetracycline andtetracycline obtained from the sample solution are not morethan each of the peak area obtained from the standard solu-tion, and the total area of the peaks, a-apooxytetracyclinehaving the relative retention time of about 2.1 with respectto oxytetracycline, b-apooxytetracycline and the peaks,which appear between a-apooxytetracycline and b-apoox-ytetracycline, is not more than the peak area of b-apoox-ytetracycline from the standard solution. The peak area of 2-acetyl-2-decarboxamide oxytetracycline, which appears afterthe principal peak, obtained from the sample solution is notmore than 4 times the peak area of 4-epioxytetracycline fromthe standard solution.Operating conditions—




Mobile phase A: Mix 60 mL of 0.33 mol/L potassium di-hydrogen phosphate TS, 100 mL of a solution oftetrabutylammonium hydrogensulfate (1 in 100), 10 mL of asolution of disodium dihydrogen ethylenediamine tetra-acetate dihydrate (1 in 2500) and 200 mL of water, and adjustthe pH to 7.5 with 2 mol/L sodium hydroxide TS. To thissolution add 30 g of t-butanol and water to make 1000 mL.

Mobile phase B: Mix 60 mL of 0.33 mol/L potassiumdihydrogen phosphate TS, 50 mL of a solution oftetrabutylammonium hydrogensulfate (1 in 100), 10 mL of asolution of disodium dihydrogen ethylenediamine tetra-

acetate dihydrate (1 in 2500) and 200 mL of water, and adjustthe pH to 7.5 with 2 mol/L sodium hydroxide TS. To thissolution add 100 g of t-butanol and water to make 1000 mL.



Mobile phaseA (z)

Mobile phaseB (z)

0 – 20 70→ 10 30→ 9020 – 35 10→ 20 90→ 80

Flow rate: 1.0 mL/minTime span of measurement: About 3.5 times as long as the

retention time of oxytetracycline after the solvent peak.System suitability—

Test for required detectability: Pipet 1 mL of 4-epioxytetracycline stock solution, and add 0.01 mol/Lhydrochloric acid TS to make exactly 200 mL. Pipet 4 mL ofthis solution, and add 0.01 mol/L hydrochloric acid TS tomake exactly 20 mL. Conˆrm that the peak area of 4-epiox-ytetracycline obtained from 20 mL of this solution is equiva-lent to 14 to 26z of that from 20 mL of the standardsolution.

System performance: Dissolve 8 mg of a-apooxytetracy-cline in 5 mL of 0.01 mol/L sodium hydroxide TS, add 0.01mol/L hydrochloric acid TS to make 100 mL, and use thissolution as a-apooxytetracycline stock solution. Mix 3 mLof the sample solution, 2 mL of 4-epioxytetracycline stocksolution, 6 mL of tetracycline hydrochloride stock solution,6 mL of b-apooxytetracycline stock solution and 6 mL of a-apooxytetracycline stock solution, and add 0.01 mol/Lhydrochloric acid TS to make 50 mL. When the procedure isrun with 20 mL of this solution under the above operatingconditions, 4-epioxytetracycline, oxytetracycline, tetracy-cline, a-apooxytetracycline and b-apooxytetracycline areeluted in this order with the resolutions between the peaks,4-epioxytetracycline and oxytetracycline, oxytetracyclineand tetracycline, and a-apooxytetracycline and b-apoox-ytetracycline being not less than 4, not less than 5 and notless than 4, respectively, and the symmetry coe‹cient of thepeak of oxytetracycline is not more than 1.3.

System repeatability: Pipet 1 mL of 4-epioxytetracyclinestock solution, and add 0.01 mol/L hydrochloric acid TS tomake exactly 200 mL. When the test is repeated 6 times with20 mL of this solution under the above operating conditions,the relative standard deviation of the peak area of 4-epiox-ytetracycline is not more than 2.0z.



Assay Weigh accurately an amount of OxytetracyclineHydrochloride and Oxytetracycline Hydrochloride Refer-ence Standard, equivalent to about 50 mg (potency), anddissolve each in diluted hydrochloric acid (1 in 100) to makeexactly 50 mL. Pipet 5 mL each of these solutions, add dilut-ed methanol (3 in 20) to make exactly 50 mL, and use these


solutions as the sample solution and the standard solution.Perform the test with exactly 20 mL each of the sample solu-tion and the standard solution as directed under the LiquidChromatography according to the following conditions, anddetermine the peak areas, AT and AS, of oxytetracycline.

Amount [mg (potency)] of oxytetracycline (C22H24N2O9)

＝WS×AT

AS× 1000

WS: Amount [mg (potency)] of OxytetracyclineHydrochloride Reference Standard



ameter and 25 cm in length, packed with strongly acidic ionexchange resin for liquid chromatography (5 mm in particlediameter).


Mobile phase: Dissolve 3.402 g of potassium dihydrogenphosphate and 9.306 g of disodium dihydrogen ethylenedia-mine tetraacetate dihydrate in 700 mL of water, add 300 mLof methanol, and adjust the pH to 4.5 with dilutehydrochloric acid.

Flow rate: Adjust the ‰ow rate so that the retention timeof oxytetracycline is about 7 minutes.System suitability—

System performance: When the procedure is run with 20mL of the standard solution under the above operating con-ditions, the theoretical plates and the symmetrical coe‹cientof the peak of oxytetracycline are not less than 1000 and notmore than 2.0, respectively.

System repeatability: When the test is repeated 6 timeswith 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peakarea of oxytetracycline is not more than 1.0z.


Peplomycin Sulfate硫酸ペプロマイシン


Peplomycin Sulfate contains not less than 865 mg(potency) per mg, calculated on the dried basis. Thepotency of Peplomycin Sulfate is expressed as mass(potency) of peplomycin (C61H88N18O21S2: 1473.59).

Description Peplomycin Sulfate occurs as a white to lightyellowish white powder.


It is hygroscopic.

Identiˆcation (1) To 4 mg of Peplomycin Sulfate add5 mL of copper (II) sulfate TS, and dissolve in water to make100 mL. Determine the absorption spectrum of this solutionas directed under the Ultraviolet-visible Spectrophotometry,and compare the spectrum with the Reference Spectrum:both spectra exhibit similar intensities of absorption at thesame wavelengths.

(2) Dissolve 10 mg each of Peplomycin Sulfate andPeplomycin Sulfate Reference Standard in 6 mL of water,add 0.5 mL of a solution of copper (II) sulfate pentahydrate(1 in 125), and use these solutions as the sample solution andthe standard solution. Perform the test with 10 mL each ofthese solutions as directed under the Liquid Chro-matography according to the following conditions: theretention time of the principal spot obtained from the sam-ple solution is the same as that from the standard soution.Operating conditions—

Detector, column, column temperature, mobile phasestock solution, mobile phase A, mobile phase B, ‰owing ofthe mobile phase, and ‰ow rate: Proceed as directed in theoperating conditions in the Purity (3).

(3) A solution of Peplomycin Sulfate (1 in 200) respondsto the Qualitative Tests (1) and (2) for sulfate.

Optical rotation [a]D20: －2 –－59(0.1 g calculated on thedried basis, 0.1 mol/L phosphate buŠer solution, pH 5.3, 10mL, 100 mm).

pH The pH of a solution obtained by dissolving 0.10 g ofPeplomycin Sulfate in 20 mL of water is between 4.5 and6.0.

Purity (1) Clarity and color of solution—Dissolve 80 mgof Peplomycin Sulfate in 4 mL of water: the solution is clearand colorless.

(2) Copper—Dissolve exactly 75 mg of Peplomycin Sul-fate in exactly 10 mL of diluted nitric acid (1 in 100), and usethis solution as the sample solution. Separately, to 5.0 mL ofStandard Copper Stock Solution add diluted nitric acid (1 in100) to make exactly 100 mL. To 3.0 mL of this solution adddiluted nitric acid (1 in 100) to make exactly 100 mL, and usethis solution as the standard solution. Perform the test withthe sample solution and the standard solution as directed un-der the Atomic Absorption Spectrophotometry according tothe following conditions: the absorbance of the sample solu-tion is not more than that of the standard solution (not morethan 200 ppm).


Lamp: Copper hollow cathode lampWavelength: 324.8 nm(3) Related substances—Dissolve about 10 mg of

Peplomycin Sulfate in 6 mL of water, add 0.5 mL of a solu-tion of copper (II) sulfate pentahydrate (1 in 125), and usethis solution as the sample solution. Perform the test with 10mL of the sample solution as directed under the Liquid Chro-matography according to the following conditions. Deter-mine the areas of the peaks, appeared after the peak of cop-per sulfate, by the automatic integration method, and calcu-


late the amounts of them by the area percentage method: thetotal amount of the peaks other than peplomycin is not morethan 7z.Operating conditions—




Mobile phase stock solution: Dissolve 0.96 g of sodium 1-pentanesulfonate and 1.86 g of disodium dihydrogenethylenediamine tetraacetate dihydrate in 1000 mL of waterand 5 mL of acetic acid (100), and adjust the pH to 4.3 withammonia TS.

Mobile phase A: A mixture of mobile phase stock solutionand methanol (9:1).

Mobile phase B: A mixture of mobile phase stock solutionand methanol (3:2).



Mobile phaseA (z)

Mobile phaseB (z)

0 – 60 100→ 0 0→ 10060 – 75 0 100

Flow rate: 1.2 mL per minute.Time span of measurement: As long as 20 minutes after

elution of peplomycin after the peak of copper sulfate.System suitability—

Test for required detectability: Measure exactly 1 mL ofthe sample solution, add water to make exactly 100 mL, anduse this solution as the solution for system suitability test.Pipet 1 mL of the solution for system suitability test, andadd water to make exactly 10 mL. Conˆrm that the peakarea of peplomycin obtained from 10 mL of this solution isequivalent to 7 to 13z of that from 10 mL of the solution forsystem suitability test.

System performance: When the procedure is run with 10mL of the sample solution under the above operating condi-tions, the number of theoretical plates and the symmetrycoe‹cient of the peak of peplomycin are not less than 30,000and not more than 2.0, respectively.

System repeatability: When the test is repeated 6 timeswith 10 mL of the sample solution under the above operatingconditions, the relative standard deviation of the peak areaof peplomycin is not more than 2.0z.

Loss on drying Not more than 3.0z (60 mg, in vacuum,phosphorus (V) oxide, 609C, 3 hours). Handle the sampleavoiding absorption of moisture.


(1) Test organism—Mycobacterium smegmatis ATCC607

(2) Agar media for seed and base layer, and for transfer-ring test organism

Glycerin 10.0 gPeptone 10.0 gMeat extract 10.0 gSodium chloride 3.0 gAgar 15.0 gWater 1000 mL

Mix all the ingredients and adjust the pH of the solutionwith sodium hydroxide TS so that it will be 6.9 to 7.1 aftersterilization.

(3) Liquid medium for suspending test organismGlycerin 10.0 gPeptone 10.0 gMeat extract 10.0 gSodium chloride 3.0 gWater 1000 mL

Mix all the ingredients and adjust the pH of the solutionwith sodium hydroxide TS so that it will be 6.9 to 7.1 aftersterilization.

(4) Preparation of agar medium of seeded layer—Inocu-late the test organism onto the slant of the agar medium fortransferring test organism, and incubate the slant at 279Cfor 40 to 48 hours. Inoculate the subcultured test organisminto 100 mL of the liquid medium for suspending test organ-ism, incubate at 25 to 279C for 5 days while shaking, and usethis suspension as the suspension of the test organism. Keepthe suspension of the test organism at a temperature of notexceeding 59C and use within 14 days. Add 0.5 mL of thesuspension of the test organism in 100 mL of the Agar medi-um for seed layer previously kept at 489C, mix thoroughly,and use this as the agar medium of seeded layer.

(5) Preparation of cylinder-agar plate—Proceed asdirected in 7. Preparation of cylinder-agar plates under theMicrobial Assay for Antibiotics with the exception of theamounts of the agar medium for base layer and the agarmedium of seeded layer to put in the Petri dish, which are5.0 mL and 8.0 mL, respectively.

(6) Standard solutions—Weigh accurately an amount ofPeplomycin Sulfate Reference Standard, equivalent to about20 mg (potency), dissolve in 0.1 mol/L phosphate buŠer so-lution, pH 6.8 to make exactly 100 mL, and use this solutionas the standard stock solution. Keep the standard stock solu-tion at a temperature not exceeding 59C, and use within 15days. Take exactly a suitable amount of the standard stocksolution before use, add 0.1 mol/L phosphate buŠer solu-tion, pH 6.8 to make solutions so that each mL contains 4 mg(potency) and 2 mg (potency), and use these solutions as thehigh concentration standard solution and the low concentra-tion standard solution, respectively.

(7) Sample solutions—Weigh accurately an amount ofPeplomycin Sulfate, equivalent to about 20 mg (potency),and dissolve in 0.1 mol/L phosphate buŠer solution, pH 6.8to make exactly 100 mL. Take exactly a suitable amount ofthis solution, add 0.1 mol/L phosphate buŠer solution, pH6.8 to make solutions so that each mL contains 4 mg (poten-


cy) and 2 mg (potency), and use these solutions as the highconcentration sample solution and the low concentrationsample solution, respectively.

Containers and storage Containers—Tight containers

Pethidine Hydrochloride塩酸ペチジン


Purity(3) Related substances—Dissolve 0.05 g of Pethidine

Hydrochloride in 20 mL of the mobile phase, and use thissolution as the sample solution. Pipet 1 mL of the sample so-lution, add the mobile phase to make exactly 100 mL, anduse this solution as the standard solution. Perform the testwith exactly 20 mL each of the sample solution and the stan-dard solution as directed under the Liquid Chromatographyaccording to the following conditions. Determine each peakarea obtained from both solutions by the automatic integra-tion method: the total area of the peaks other than that ofpethidine from the sample solution is not larger than thepeak area of perthidine from the standard solution.Operating conditions—




Mobile phase: Dissolve 2.0 g of sodium lauryl sulfate in1000 mL of diluted phosphoric acid (1 in 1000), adjust thepH to 3.0 with sodium hydroxide TS, and to 550 mL of thissolution add 450 mL of acetonitrile.

Flow rate: Adjust the ‰ow rate so that the retention timeof pethidine is about 7 minutes.

Time span of measurement: About 2 times as long as theretention time of pethidine after the solvent peak.System suitability—

Test for required detection: To exactly 2 mL of the stan-dard solution add the mobile phase to make exactly 20 mL.Conˆrm that the peak area of pethidine obtained from 20 mLof this solution is equivalent to 5 to 15z of that of pethidineobtained from 20 mL of the standard solution.

System performance: To 2 mL each of the sample solutionand a solution of isoamyl parahydroxybenzoate in the mo-bile phase (1 in 50,000) add the mobile phase to make 10 mL.When the procedure is run with 20 mL of this solution ac-cording to the above operating conditions, pethidine andisoamyl parahydroxybenzoate are eluted in this order withthe resolution between these peaks being not less than 2.0.

System repeatability: When the test is repeated 6 timeswith 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak

area of pethidine is not more than 2.0z.

Add the following:

Phenethicillin Potassiumフェネチシリンカリウム

C17H19KN2O5S: 402.51Monopotassium (2S,5R,6R )-3,3-dimethyl-7-oxo-6-[(2RS )-2-phenoxypropanoylamino]-4-thia-1-azabicyclo[3.2.0]heptane-2-carboxylate [132-93-4 ]

Phenethicillin Potassium contains not less than 1400units per mg, calculated on the dried basis. The poten-cy of Phenethicillin Potassium is expressed as unitbased on the amount of phenethicillin potassium(C17H19KN2O5S). 1 unit of Phenethicillin Potassium isequivalent to 0.68 mg of phenethicillin potassium(C17H19KN2O5S).

Description Phenethicillin Potassium occurs as a white tolight yellowish white crystalline powder.


Identiˆcation (1) Determine the absorption spectrum ofa solution of Phenethicillin Potassium (1 in 5000) as directedunder the Ultraviolet-visible Spectrophotometry, and com-pare the spectrum with the Reference Spectrum: both spec-tra exhibit similar intensities of absorption at the samewavelengths.

(2) Determine the infrared absorption spectrum ofPhenethicillin Potassium as directed in the potassiumbromide disk method under the Infrared Spectrophotomet-ry, and compare the spectrum with the Reference Spectrum:both spectra exhibit similar intensities of absorption at thesame wave numbers.

(3) Phenethicillin Potassium responds to the QualitativeTest (1) for potassium salt.

Optical rotation [a]D20: ＋217 –＋2449(1 g calculated onthe dried basis, phosphate TS, 100 mL, 100 mm).

L-a-Phenethicillin potassium Dissolve about 50 mg ofPhenethicillin Potassium in the mobile phase to make ex-actly 50 mL, and use this solution as the sample solution.Perform the test with 10 mL of the sample solution as direct-ed under the Liquid Chromatography according to the fol-lowing conditions, and determine the peak areas, AD andAL, of D-a-phenethicillin and L-a-phenethicillin by the auto-matic integration method: AL/(AD＋ AL) is between 0.50and 0.70.Operating conditions—

Detector: An ultraviolet absorption photometer





Mobile phase: Adjust the pH of a mixture of a solution ofdiammonium hydrogen phosphate (1 in 150) and acetonitrile(41:10) to 7.0 with phosphoric acid.

Flow rate: Adjust the ‰ow rate so that the retention timeof L-a-phenethicillin is about 25 minutes.System suitability—

System performance: When the procedure is run with 10mL of the sample solution under the above operating condi-tions, D-a-phenethicillin and L-a-phenethicillin are eluted inthis order with the resolution between these peaks being notless than 1.5.

System repeatability: When the test is repeated 6 timeswith 10 mL of the sample solution under the above operatingconditions, the relative standard deviation of the peak areaof L-a-phenethicillin is not more than 2.0z.

Purity (1) Heavy metals—Proceed with 1.0 g ofPhenethicillin Potassium according to Method 2, and per-form the test. Prepare the control solution with 1.0 mL ofStandard Lead Solution (not more than 10 ppm).

(2) Arsenic—Prepare the test solution with 1.0 g ofPhenethicillin Potassium according to Method 4 and, per-form the test (not more than 2 ppm).

(3) Related substances—Dissolve 50 mg of Phenethicil-lin Potassium in 50 mL of the mobile, and use this solutionas the sample solution. Pipet 1 mL of the sample solution,add the mobile phase to make exactly 100 mL, and use thissolution as the standard solution. Perform the test withexactly 10 mL each of the sample solution and the standardsolution as directed under the Liquid Chromatography ac-cording to the following conditions, and determine eachpeak area by the automatic integration method: the total ofthe peak areas other than D-a-phenethicillin and L-a-phenethicillin obtained from the sample solution is not morethan 5 times the total of the peak areas of D-a-phenethicillinand L-a-phenethicillin from the standard solution.Operating conditions—

Detector, column, column temperature, mobile phase,and ‰ow rate: Proceed as directed in the operating condi-tions in the L-a-Phenethicillin potassium.

Time span of measurement: About 1.5 times as long as theretention time of L-a-phenethicillin.System suitability—

Test for required detectability: Measure exactly 2 mL ofthe standard solution, and add the mobile phase to make ex-actly 10 mL. Conˆrm that the peak area of L-a-phenethicil-lin obtained from 10 mL of this solution is equivalent to 14 to26z of that from 10 mL of the standard solution.

System performance, and system repeatability: Proceed asdirected in the system suitability in the L-a-Phenethicillinpotassium.

Loss on drying Not more than 1.0z (0.1 g, in vacuum,609C, 3 hours).

Assay Weigh accurately an amount of PhenethicillinPotassium and dried L-Phenethicillin Potassium ReferenceStandard, equivalent to about 40,000 units, dissolve each inphosphate buŠer solution, pH 6.0 to make exactly 20 mL,and use these solutions as the sample solution and the stan-dard solution, respectively. Pipet 2 mL each of these solu-tions in 100-mL glass-stoppered ‰asks, add 2.0 mL of sodi-um hydroxide TS to them, and allow to stand for exactly 15minutes. To them add 2.0 mL of diluted hydrochloric acid (1in 10) and exactly 10 mL of 0.005 mol/L iodine VS, and al-low them to stand for exactly 15 minutes. Add 0.2 – 0.5 mLof starch TS, and titrate with 0.01 mol/L sodium thiosulfateVS until the color of the solution disappears. Separately, toexactly 2 mL each of the sample solution and the standardsolution add exactly 10 mL of 0.005 mol/L iodine VS, thenproceed in the same manner as above without allowing tostand for 15 minutes as a blank determination, and makeany necessary correction. Determine the volumes, VT andVS, of 0.005 mol/L iodine VS consumed in the sample solu-tion and the standard solution.

Amount (unit) of C17H19KN2O5S＝WS×VT

VS

WS: Amount (unit) of L-Phenethicillin Potassium Refer-ence Standard



Phytonadioneフィトナジオン


Description Phytonadione is a clear yellow to orange-yel-low, viscous liquid.

It is miscible with isooctane.It is sparingly soluble in ethanol (99.5), and practically in-

soluble in water.It decomposes gradually and changes to a red-brown by

light.Speciˆc gravity d 20

20: about 0.967


Identiˆcation (1) Determine the absorption spectrum ofa solution of Phytonadione in isooctane (1 in 100,000) asdirected under the Ultraviolet-visible Spectrophotometry,and compare the spectrum with the Reference Spectrum 1:both spectra exhibit similar intensities of absorption at thesame wavelengths. Separately, determine the absorptionspectrum of a solution of Phytonadione in isooctane (1 in10,000) as directed under the Ultraviolet-visible Spec-trophotometry, and compare the spectrum with the Refer-ence Spectrum 2: both spectra exhibit similar intensities ofabsorption at the same wavelengths.

(2) Determine the infrared absorption spectrum ofPhytonadione as directed in the liquid ˆlm method under theInfrared Spectrophotometry, and compare the spectrumwith the Reference Spectrum: both spectra exhibit similar in-tensities of absorption at the same wave numbers.


Purity (1) Ratio of absorbances—Determine the absor-bances, A1, A2 and A3, of a solution of Phytonadione inisooctane (1 in 100,000) at 248.5 nm, 253.5 nm and 269.5nm, respectively: the ratio A2/A1 is between 0.69 and 0.73,and the ratio A2/A3 is between 0.74 and 0.78. Determine theabsorbances, A4 and A5, of a solution of Phytonadione inisooctane (1 in 10,000) at 284.5 nm and 326 nm, respectively:the ratio A4/A5 is between 0.28 and 0.34.

(2) Heavy metals—Carbonize 1.0 g of Phytonadione bygentle heating. Cool, add 10 mL of a solution of magnesiumnitrate hexahydrate in ethanol (95) (1 in 10), and ignite theethanol to burn. Cool, add 1 mL of sulfuric acid, proceedaccording to Method 4, and perform the test. Prepare thecontrol solution with 2.0 mL of Standard Lead Solution (notmore than 20 ppm).

(3) Menadione—Dissolve 0.020 g of Phytonadione in0.5 mL of a mixture of water and ethanol (95) (1:1), add 1drop of a solution of 3-methyl-1-phenyl-5-pyrazolone inethanol (95) (1 in 20) and 1 drop of ammonia solution (28),and allow to stand for 2 hours: no blue-purple colordevelops.

Add the following:

PimaricinNatamycin

ピマリシン

C33H47NO13: 665.73(1R *,3S *,5R *,7R *,8E,12R *,14E,16E,18E,20E,22R *,24S *,25R *,26S *)-22-(3-Amino-3,6-dideoxy-b-D-mannopyranosyloxy)-1,3,26-trihydroxy-12-methyl-10-oxo-6,11,28-trioxatricyclo[22.3.1.05,7]octacosa-8,14,16,18,20-pentaene-25-carboxylic acid [7681-93-8 ]

Pimaricin contains not less than 900 mg (potency)per mg, calculated on the anhydrous basis. The poten-cy of Pimaricin is expressed as mass (potency) ofpimaricin (C33H47NO13).

Description Pimaricin occurs as white to yellowish whitecrystalline powder.

It is slightly soluble in methanol and in acetic acid (100),and practically insoluble in water and in ethanol (99.5).

Identiˆcation (1) To 3 mg of Pimaricin add 1 mL ofhydrochloric acid, and mix: a blue-purple color appears.

(2) Dissolve 5 mg of Pimaricin in a solution of aceticacid (100) in methanol (1 in 100) to make 1000 mL. Deter-mine the absorption spectrum of this solution as directed un-der the Ultraviolet-visible Spectrophotometry, and comparethe spectrum with the Reference Spectrum or the spectrumof a solution of Pimaricin Reference Standard prepared inthe same manner as the sample solution: both spectra exhibitsimilar intensities of absorption at the same wavelengths.

Optical rotation [a]D20: ＋243 –＋2599(0.1 g, acetic acid(100), 25 mL, 100 mm).

Purity (1) Heavy metals—Proceed with 1.0 g of Pimari-cin according to Method 4, and perform the test. Prepare thecontrol solution with 3.0 mL of Standard Lead Solution (notmore than 30 ppm).

(2) Related substances—Dissolve 20 mg of Pimaricin inmethanol to make 100 mL, and use this solution as the sam-ple solution. Perform the test with 10 mL of the sample solu-tion as directed under the Liquid Chromatography accord-ing to the following conditions, and determine the total areaof the peaks other than pimaricin by the automatic integra-tion method: not more than 4.0z.






Mobile phase: Dissolve 1.0 g of ammonium acetate in1000 mL of a mixture of water, methanol and tetrahydrofu-ran (47:44:2).

Flow rate: Adjust the ‰ow rate so that the retention timeof pimaricin is about 10 minutes.

Time span of measurement: About 3 times as long as theretention time of pimaricin.System suitability—

Test for required detectability: Measure exactly 1 mL ofthe sample solution, add methanol to make exactly 100 mL,and use this solution as the solution for system suitabilitytest. Pipet 1 mL of the solution for system suitability test,and add methanol to make exactly 10 mL. Conˆrm that thepeak area of pimaricin obtained from 10 mL of this solutionis equivalent to 7 to 13z of that from 10 mL of the solutionfor system suitability test.

System performance: When the procedure is run with10 mL of the solution for system suitability test under theabove operating conditions, the number of theoretical platesand the symmetry coe‹cient of the peak of pimaricin are notless than 1500 and not more than 2.0, respectively.

System repeatability: When the test is repeated 6 timeswith 10 mL of the solution for system suitability test underthe above operating conditions, the relative standard devia-tion of the peak area of pimaricin is not more than 2.0z.

Water Between 6.0z and 9.0z (0.2 g, volumetric titra-tion, direct titration).

Assay Weigh accurately an amount of Pimaricin andPimaricin Reference Standard, equivalent to about 25 mg(potency), and dissolve each in methanol to make exactly100 mL. Pipet 2 mL each of these solution, add a solution ofacetic acid (100) in methanol (1 in 100) to make exactly 100mL, and use these solutions as the sample solution and thestandard solution. Determine the absorbances at 295.5 nm,AT1 and AS1, at 303 nm, AT2 and AS2, and at 311 nm, AT3

and AS3, of the sample solution and the standard solution asdirected under the Ultraviolet-visible Spectrophotometry.

Amount [mg (potency)] of C33H47NO13

＝WS×

AT2－AT1＋ AT3

2

AS2－AS1＋ AS3

2

× 1000

WS: Amount [mg (potency)] of Pimaricin ReferenceStandard

Containers and storage Containers—Tight containers.Storage—Light resistant.

Pipemidic Acid Trihydrateピペミド酸三水和物


Pipemidic Acid Trihydrate, when dried, containsnot less than 98.5z and not more than 101.0z ofC14H17N5O3 (mol. wt.: 303.32).


Description Pipemidic Acid Trihydrate occurs as a paleyellow, crystalline powder.

It is freely soluble in acetic acid (100), very slightly solublein water and in ethanol (99.5).

It dissolves in sodium hydroxide TS.It is gradually colored by light.Melting point: about 2509C (with decomposition).


Identiˆcation (1) Dissolve 0.1 g of Pipemidic Acid Trihy-drate in 20 mL of sodium hydroxide TS, and dilute withwater to make 200 mL. To 1 mL of the solution add water tomake 100 mL. Determine the absorption spectrum of thesolution as directed under the Ultraviolet-visible Spec-trophotometry, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of ab-sorption at the same wavelengths.

(2) Determine the infrared absorption spectrum ofPipemidic Acid Trihydrate as directed in the potassiumbromide disk method under the Infrared Spectrophotomet-ry, and compare the spectrum with the Reference Spectrum:both spectra exhibit similar intensities of absorption at thesame wave numbers.


Assay Weigh accurately about 0.3 g of Pipemidic Acid Tri-hydrate, previously dried, dissolve in 40 mL of acetic acid(100), and titrate with 0.1 mol/L perchloric acid VS (poten-tiometric titration). Perform a blank determination, andmake any necessary correction.

Each mL of 0.1 mol/L perchloric acid VS＝ 30.33 mg of C14H17N5O3

Piperacillin Sodiumピペラシリンナトリウム


Description Piperacillin Sodium occurs as a white powderor mass.

It is very soluble in water, freely soluble in methanol andin ethanol (95), and practically insoluble in acetonitrile.


Add the following:

Pirarubicinピラルビシン

C32H37NO12: 627.64(2S,4S )-4-｛3-Amino-2,3,6-trideoxy-4-O-[(2R )-tetrahydropyran-2-yl]-a-L-lyxo-hexopyranosyloxy｝-1,2,3,4-tetrahydro-2,5,12-trihydroxy-2-hydroxyacetyl-7-methoxynaphthacene-6,11-dione [72496-41-4 ]

Pirarubicin contains not less than 950 mg (potency)per mg, calculated on the anhydrous basis. The poten-cy of Pirarubicin is expressed as mass (potency) ofpirarubicin (C32H37NO12).

Description Pirarubicin occurs as a red-orange crystallinepowder.

It is soluble in chloroform, very slightly soluble in acetoni-trile, in methanol and in ethanol (99.5), and practically in-soluble in water.

Identiˆcation (1) Dissolve 10 mg of Pirarubicin in 80 mLof methanol and 6 mL of diluted hydrochloric acid (1 in5000), and add water to make 100 mL. To 10 mL of this so-lution add diluted methanol (4 in 5) to make 100 mL. Deter-mine the absorption spectrum of this solution as directed un-der the Ultraviolet-visible Spectrophotometry, and comparethe spectrum with the Reference Spectrum or the spectrumof a solution of Pirarubicin Reference Standard prepared inthe same manner as the sample solution: both spectra exhibitsimilar intensities of absorption at the same wavelengths.

(2) Dissolve 5 mg each of Pirarubicin and PirarubicinReference Standard in 5 mL of chloroform, and use thesesolutions as the sample solution and the standard solution.Perform the test with these solutions as directed under theThin-layer Chromatography. Spot 5 mL each of the samplesolution and the standard solution on a plate of silica gel forthin-layer chromatography. Develop the plate with a mix-ture of chloroform and methanol (5:1) to a distance of about10 cm, and air-dry the plate. Examine the spots with thenecked eye: the principal spot obtained from the samplesolution and the spot from the standard solution show a red-orange color and the same Rf value.

Optical rotation [a]D20:＋195 –＋2159(10 mg, chloroform,10 mL, 100 mm).

Purity (1) Clarity and color of solution—Dissolve 10 mgof Pirarubicin in 10 mL of 0.01 mol/L hydrochloric acid TS:

the solution is clear and red.(2) Heavy metals—Proceed with 1.0 g of Pirarubicin ac-

cording to Method 2, and perform the test. Prepare the con-trol solution with 2.0 mL of Standard Lead Solution (notmore than 20 ppm).

(3) Related substances—Dissolve 10 mg of Pirarubicinin 20 mL of the mobile phase, and use this solution as thesample solution. Pipet 1 mL of the sample solution, add themobile phase to make exactly 200 mL, and use this solutionas the standard solution. Perform the test with exactly 20 mLeach of the sample solution and the standard solution asdirected under the Liquid Chromatography according to thefollowing conditions, and determine each peak area by theautomatic integration method: the peak area of doxorubi-cin, having the relative retention time of about 0.45 withrespect to pirarubicin, and the area of the peak, having therelative retention time of about 1.2 with respect to pirarubi-cin, obtained from the sample solution are not more than thepeak area of pirarubicin from the standard solution, respec-tively, and the sum of the areas of the peaks, having the rela-tive retention times of about 1.9 and about 2.0 with respectto pirarubicin, from the sample solution is not more than 5times the peak area of pirarubicin from the standard solu-tion. For these calculations, use the peak area for doxorubi-cin after multiplying by the response factor 0.94 and the areafor the two peaks, having the relative retention times ofabout 1.9 and about 2.0, after multiplying by their responsefactors, 1.09, respectively.Operating conditions—


Time span of measurement: About 4 times as long as theretention time of pirarubicin.System suitability—

Test for required detectability: Measure exactly 2 mL ofthe standard solution, and add the mobile phase to make ex-actly 10 mL. Conˆrm that the peak area of pirarubicin ob-tained from 20 mL of this solution is equivalent to 14 to 26zof that from 20 mL of the standard solution.



Assay Weigh accurately an amount of Pirarubicin andPirarubicin Reference Standard, equivalent to about 10 mg(potency), and dissolve in the mobile phase to make exactly10 mL. Pipet 5 mL of these solutions, add exactly 5 mL ofthe internal standard solution, and use these solutions as thesample solution and the standard solution. Perform the testwith 20 mL each of the sample solution and the standard so-lution as directed under the Liquid Chromatography accord-ing to the following conditions, and determine the ratios, QT

and QS, of the peak area of pirarubicin to that of the internalstandard.


Amount [mg (potency)] of C32H37NO12＝WS×QT

QS× 1000

WS: Amount [mg (potency)] of Pirarubicin ReferenceStandard

Internal standard solution—A solution of 2-naphthol in themobile phase (1 in 1000).Operating conditions—




Mobile phase: A mixture of 0.05 mol/L ammonium for-mate buŠer solution, pH 4.0 and acetonitrile (3:2).

Flow rate: Adjust the ‰ow rate so that the retention timeof pirarubicin is about 7 minutes.System suitability—

System performance: When the procedure is run with 20mL of the standard solution under the above operating con-ditions, pirarubicin and the internal standard are eluted inthis order with the resolution between these peaks being notless than 9.

System repeatability: When the test is repeated 6 timeswith 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratiosof the peak area of pirarubicin to that of the internal stan-dard is not more than 1.0z.


Pivmecillinam Hydrochloride塩酸ピブメシリナム


Pivmecillinam Hydrochloride contains not less than630 mg (potency) per mg, calculated on the anhydrousbasis. The potency of Pivmecillinam Hydrochlorideis expressed as mass (potency) of pivmecillinam(C21H33N3O5S: 439.57).

Description Pivmecillinam Hydrochloride occurs as awhite to yellowish white crystalline powder.

It is very soluble in methanol and in acetic acid (100), free-ly soluble in water and in ethanol (99.5), and soluble inacetonitrile.

Identiˆcation (1) Determine the infrared absorptionspectrum of Pivmecillinam Hydrochloride as directed in thepotassium bromide disk method under the Infrared Spec-trophotometry, and compare the spectrum with the Refer-ence Spectrum or the spectrum of Pivmecillinam Hydrochlo-ride Reference Standard: both spectra exhibit similar intensi-ties of absorption at the same wave numbers.

(2) Dissolve 0.5 g of Pivmecillinam Hydrochloride in10 mL of water, and add 1 mL of dilute nitric acid and 1drop of silver nitrate TS: a white precipitate is formed.

Optical rotation [a]D20: ＋200 –＋2209(1 g calculated onthe anhydrous basis, water, 100 mL, 100 mm).

Purity (1) Heavy metals—To 1.0 g of PivmecillinamHydrochloride in a crucible add 10 mL of a solution of mag-nesium nitrate hexahydrate in ethanol (95) (1 in 10), ˆre theethanol to burn, and heat gradually to incinerate. If a car-bonized substance remains, moisten with a small amount ofnitric acid, and ignite to incinerate. Cool, add 3 mL ofhydrochloric acid to the residue, dissolve by warming on awater bath, and heat to dryness. To the residue add 10 mL ofwater, and dissolve by warming on a water bath. After cool-ing, adjust the pH to 3 to 4 with ammonia TS, add 2 mL ofdilute acetic acid, ˆlter if necessary, and wash the crucibleand the ˆlter with 10 mL of water. Put the ˆltrate and thewashings to a Nessler tube, add water to make 50 mL, anduse this solution as the test solution. Prepare the control so-lution in the same manner as the test solution with 2.0 mL ofStandard Lead Solution (not more than 20 ppm).

(2) Arsenic—Prepare the test solution with 1.0 g of Piv-mecillinam Hydrochloride according to Method 4, and per-form the test (not more than 2 ppm).

(3) Related substances—Dissolve 0.05 g of Pivmecillin-am Hydrochloride in 4.0 mL of a mixture of acetonitrile andacetic acid (100) (97:3), and use this solution as the samplesolution. Separately, dissolve 2.0 mg of PivmecillinamHydrochloride Reference Standard in 4.0 mL of water, anduse this solution as the standard solution. Perform the testwith these solutions as directed under the Thin-layer Chro-matography. Spot 2 mL of the standard solution on a plateof silica gel for thin-layer chromatography, allow to standfor 30 minutes, then spot 2 mL of the sample solution on theplate. Immediately, develop the plate with a mixture of ace-tone, water and acetic acid (100) (10:1:1) to a distance ofabout 12 cm, and air-dry the plate. Allow the plate to standfor 10 minutes in iodine vapor: the spot from the sample so-lution appeared at the position corresponding to the spot ob-tained from the standard solution is not larger and not moreintense than the spot from the standard solution, and anyspot other than the principal spot and the above spot is notobservable.


Assay Weigh accurately an amount of PivmecillinamHydrochloride and Pivmecillinam Hydrochloride ReferenceStandard, equivalent to about 20 mg (potency), dissolve in asuitable amount of the mobile phase, add exactly 10 mL ofthe internal standard solution and the mobile phase to make100 mL, and use these solutions as the sample solution andthe standard solution, respectively. Perform the test with 10mL each of the sample solution and the standard solution asdirected under the Liquid Chromatography according to thefollowing conditions, and determine the ratios, QT and QS,of the peak area of pivmecillinam to that of the internalstandard.


Amount [mg (potency)] of pivmecillinam (C21H33N3O5S)

＝WS×QT

QS× 1000

WS: Amount [mg (potency)] of Pivmecillinam Hydro-chloride Reference Standard

Internal standard solution—A solution of diphenyl in themobile phase (1 in 12,500).Operating conditions—




Mobile phase: Dissolve 0.771 g of ammonium acetate inabout 900 mL of water, adjust the pH to 3.5 with acetic acid(100), and add water to make 1000 mL. To 400 mL of thissolution add 600 mL of acetonitrile.

Flow rate: Adjust the ‰ow rate so that the retention timeof pivmecillinam is about 6.5 minutes.System suitability—

System performance: When the procedure is run with 10mL of the standard solution under the above operating con-ditions, pivmecillinam and the internal standard are elutedin this order with the resolution between these peaks beingnot less than 4.

System repeatability: When the test is repeated 6 timeswith 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratiosof the peak area of pivmecillinam to that of the internalstandard is not more than 1.0z.


Polymixin B Sulfate硫酸ポリミキシン B


Polymixin B Sulfate contains not less than 6500units per mg, calculated on the dried basis. The poten-cy of Polymixin B Sulfate is expressed as mass unit ofpolymixin B (C55-56H96-98N16O13). One unit of Poly-mixin B Sulfate is equivalent to 0.129 mg of polymixinB sulfate (C55-56H96-98N16O13.1-2H2SO4).

Description Polymixin B Sulfate occurs as a white to yel-low-brown powder.


Identiˆcation (1) To 5 mL of a solution of Polymixin BSulfate (1 in 10) add 5 mL of a solution of sodium hydroxide(1 in 10), add 5 drops of a solution of copper (II) sulfate pen-

tahydrate (1 in 100) while shaking: a purple color develops.(2) Transfer 5 mg each of Polymixin B Sulfate and Poly-

mixin B Sulfate Reference Standard separately in two glassstoppered test tubes, add 1 mL of diluted hydrochloric acid(1 in 2), stopper the tube, heat at 1359C for 5 hours, thenheat to dryness on a water bath, and keep the heating untilno more hydrochloric acid odor is evolved. Dissolve theresidue in 0.5 mL of water, and use these solutions as thesample solution and the standard solution (1). Separately,dissolve 20 mg each of L-leucine, L-threonine, phenylalanineand L-serine separately in 10 mL of water, and use these so-lutions as the standard solution (2), the standard solution(3), the standard solution (4) and the standard solution (5),respectively. Perform the test with these solutions as directedunder the Thin-layer Chromatography. Spot 3 mL each ofthe sample solution, the standard solution (1), the standardsolution (2), the standard solution (3), the standard solution(4) and the standard solution (5) on a plate of silica gel forthin-layer chromatography, and expose the plate to a satu-rated vapor of the developing solvent for 15 hours. Developthe plate with a mixture of phenol and water (3:1) to a dis-tance of about 13 cm while without exposure to light, anddry the plate at 1109C for 5 minutes. Spray evenly nin-hydrin-acetic acid TS on the plate, and heat at 1109C for 5minutes: Rf value of each spot obtained from the sample so-lution is the same with Rf value of the corresponding spotsfrom the standard solution (1). Each of the spots from thesample solution appears at the position corresponding toeach of the spots from the standard (2), (3) and (4), but notappears at the position corresponding to the spot from thestandard solution (5).

(3) A solution of Polymixin B Sulfate (1 in 20) respondsto the Qualitative Tests for sulfate.

Optical rotation [a]D20: －78 –－909(0.5 g calculated onthe dried basis, water, 25 mL, 100 mm).

pH The pH of a solution obtained by dissolving 1.0 g ofPolymixin B Sulfate in 50 mL of water is between 5.0 and7.0.

Phenylalanine Weigh accurately about 0.375 g of Polymix-in B Sulfate, dissolve in 0.1 mol/L hydrochloric acid TS tomake exactly 100 mL. Determine absorbances, A1, A2, A3,A4 and A5, of this solution at 252 nm, at 258 nm, at 264 nm,at 280 nm and at 300 nm, respectively, as directed under theUltraviolet-visible Spectrophotometry, and calculate theamount of phenylalanine by the following equation: theamount of phenylalanine calculated on the dried basis is notless than 9z and not more than 12z.

Amount (z) of phenylalanine

＝A2－0.5A1＋0.5A3－1.8A4＋0.8A5

WT× 9.4787

WT: Amount (g) of the sample, calculated on the driedbasis

Purity Heavy metals—Proceed with 1.0 g of Polymixin BSulfate according to Method 2, and perform the test. Pre-pare the control solution with 2.0 mL of Standard Lead


Solution (not more than 20 ppm).




(1) Test organism—Escherichia coli NIHJ(2) Agar media for seed and base layer

Peptone 10.0 gMeat extract 3.0 gSodium chloride 30.0 gAgar 20.0 gWater 1000 mL


(3) Standard solutions—Weigh accurately an amount ofPolymixin B Sulfate Reference Standard, equivalent toabout 200,000 units, dissolve in phosphate buŠer solution,pH 6.0 to make exactly 20 mL, and use this solution as thestandard stock solution. Keep the standard stock solution atnot exceeding 59C and use within 14 days. Take exactly asuitable amount of the standard stock solution before use,add phosphate buŠer solution, pH 6.0 to make solutions sothat each mL contains 4000 units and 1000 units, and usethese solutions as the high concentration standard solutionand the low concentration standard solution, respectively.

(4) Sample solutions—Weigh accurately an amount ofPolymixin B Sulfate, equivalent to about 200,000 units, anddissolve in phosphate buŠer solution, pH 6.0 to make ex-actly 20 mL. Take exactly a suitable amount of this solution,add phosphate buŠer solution, pH 6.0 to make solutions sothat each mL contains 4000 units and 1000 units, and usethese solutions as the high concentration sample solutionand the low concentration sample solution, respectively.


Potassium Clavulanateクラブラン酸カリウム


Potassium Clavulanate contains not less than 810 mg(potency) per mg, calculated on the anhydrous basis.The potency of Potassium Clavulanate is expressed asmass (potency) of clavularic acid (C8H9NO5: 199.16).


Identiˆcation (1) To 1 mL of a solution of PotassiumClavulanate (1 in 50,000) add 5 mL of imidazole TS, andwarm in a water bath at 309C for 12 minutes. After cooling,determine the absorption spectrum of this solution as direct-

ed under the Ultraviolet-visible Spectrophotometry, andcompare the spectrum with the Reference Spectrum: bothspectra exhibit similar intensities of absorption at the samewavelengths.


Optical rotation [a]D20: ＋53 –＋639(0.5 g calculated onthe anhydrous basis, water, 50 mL, 100 mm).

Delete the pH.


Purity(3) Related substances—Dissolve 0.10 g of Potassium

Clavulanate in 10 mL of the mobile phase A, and use this so-lution as the sample solution. Pipet 1 mL of the sample solu-tion, add the mobile phase to make exactly 100 mL, and usethis solution as the standard solution. Perform the test withexactly 20 mL each of the sample solution and the standardsolution as directed under the Liquid Chromatography ac-cording to the following conditions, and determine eachpeak area by the automatic integration method: the area ofeach peak other than clavulanic acid from the sample solu-tion is not more than the peak area of clavulanic acid fromthe standard solution, and the total area of the peaks otherthan clavulanic acid from the sample solution is not morethan 2 times of the peak area of clavulanic acid from thestandard solution.Operating conditions—




Mobile phase A: Adjust the pH of 0.05 mol/L sodium di-hydrogen phosphate TS to 4.0 with phosphoric acid.

Mobile phase B: A mixture of the mobile phase A andmethanol (1:1).



Mobile phaseA (z)

Mobile phaseB (z)

0 – 4 100 04 – 15 100→ 0 0→ 100

15 – 25 0 100

Flow rate: 1.0 mL per minute.Time span of measurement: About 6 times as long as the

retention time of clavulanic acid.System suitability—

Test for required detectability: Pipet 1 mL of the standardsolution, and add the mobile phase A to make exactly 10


mL. Conˆrm that the peak area of clavulanic acid obtainedfrom 20 mL of this solution is equivalent to 7 to 13z of thatfrom 20 mL of the standard solution.

System performance: Dissolve 10 mg each of PotassiumClavulanate and Amoxycillin Reference Standard in 100 mLof the mobile phase A. When the procedure is run with 20mL of this solution under the above operating conditions,clavulanic acid and amoxycillin are eluted in this order withthe resolution between these peaks being not less than 8 andthe number of theoretical plates of the peak of clavulanicacid is not less than 2500 steps.

System repeatability: When the test is repeated 3 timeswith 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peakarea of clavulanic acid is not more than 2.0z.

Povidone-Iodineポビドンヨード


Description Povidone-Iodine occurs as a dark red-brownpowder. It has a faint, characteristic odor.

It is freely soluble in water and in ethanol (99.5).The pH of a solution obtained by dissolving 1.0 g of

Povidone-Iodine in 100 mL of water is between 1.5 and 3.5.


Purity(3) Arsenic—Prepare the test solution with 1.0 g of

Povidone-Iodine according to Method 4, and perform thetest (not more than 2 ppm).


Assay(2) Nitrogen—Weigh accurately about 20 mg of

Povidone-Iodine, and perform the test as directed under theNitrogen Determination.

Prednisolone Acetate酢酸プレドニゾロン


Assay Dissolve about 10 mg each of Prednisolone Acetateand Prednisolone Acetate Reference Standard, previouslydried and accurately weighed, in 60 mL each of methanol,add exactly 2 mL each of the internal standard solution, thenadd methanol to make 100 mL, and use these solutions asthe sample solution and the standard solution. Perform thetext with 10 mL each of the sample solution and the standardsolution as directed under the Liquid Chromatography ac-cording to the following conditions, and calculate the ratios,QT and QS, of the peak height of prednisolone acetate to

that of the internal standard.


QS

WS: Amount (mg) of Prednisolone Acetate ReferenceStandard






of prednisolone acetate is about 10 minutes.System suitability—

System performance: When the procedure is run with 10mL of the standard solution under the above operating con-ditions, prednisolone acetate and the internal standard areeluted in this order with the resolution between these peaksbeing not less than 10.

System repeatability: When the test is repeated 6 timeswith 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratiosof the peak height of prednisolone acetate to that of the in-ternal standard is not more than 1.0z.

Prednisolone Sodium Succinatefor Injection注射用コハク酸プレドニゾロンナトリウム


Assay Take a quantity of sealed containers of Predniso-lone Sodium Succinate for Injection, equivalent to about0.10 g of prednisolone (C21H28O5), and dissolve the contentsin a suitable amount of diluted methanol (1 in 2), and trans-fer to a 100-mL volumetric ‰ask. Wash each container withdiluted methanol (1 in 2), collect the washings in the volu-metric ‰ask, and add diluted methanol (1 in 2) to makevolume. Pipet 4 mL of this solution, add diluted methanol (1in 2) to make exactly 50 mL. Pipet 5 mL of this solution, addexactly 5 mL of the internal standard solution, mix, and usethis solution as the sample solution. Separately, weigh ac-curately about 25 mg of Prednisolone Succinate ReferenceStandard, previously dried in a desiccator for 6 hours (invacuum, phosphorus (V) oxide, 609C), dissolve in methanolto make exactly 25 mL. Pipet 5 mL of this solution, adddiluted methanol (1 in 2) to make exactly 50 mL. Pipet 5 mLof this solution, add exactly 5 mL of the internal standardsolution, mix, and use this solution as the standard solution.


Perform the test with 10 mL of the sample solution and thestandard solution as directed under the Liquid Chro-matography according to the following conditions, and cal-culate the ratios, QT and QS, of the peak area of predniso-lone succinate to that of the internal standard.

Amount (mg) of prednisolone sodium succinate(C25H31NaO8)

＝WS×QT

QS× 5× 1.0477

Amount (mg) of prednisolone (C21H28O5)

＝WS×QT

QS× 5× 0.7827

WS: Amount (mg) of Prednisolone Succinate ReferenceStandard

Internal standard solution—A solution of propyl para-hydroxybenzoate in diluted methanol (1 in 2) (1 in 25,000).Operating conditions—




Mobile phase: Dissolve 0.32 g of tetra n-butylammoniumbromide, 3.22 g of disodium hydrogen phosphate 12-waterand 6.94 g of potassium dihydrogen phosphate in 1000 mLof water. To 840 mL of this solution add 1160 mL ofmethanol.

Flow rate: Adjust the ‰ow rate so that the retention timeof prednisolone succinate is about 15 minutes.System suitability—

System performance: When the procedure is run with 10mL of the standard solution under the above operating con-ditions, prednisolone succinate and the internal standard areeluted in this order with the resolution between these peaksbeing not less than 6.

System repeatability: When the test is repeated 6 timeswith 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratiosof the peak area of prednisolone succinate to that of the in-ternal standard is not more than 1.0z.

Primidoneプリミドン


Purity(3) 2-Ethyl-2-phenylmalonediamide—Dissolve 0.10 g of

Primidone in 2 mL of pyridine, add exactly 2 mL of the in-ternal standard solution, then add 1 mL of bis-trimethyl silylacetamide, shake well, and heat at 1009C for 5 minutes.Cool, add pyridine to make 10 mL, and use this solution as

the sample solution. Separately, dissolve 50 mg of 2-ethyl-2-phenylmalonediamide in pyridine to make exactly 100 mL.Pipet 2 mL of this solution, add exactly 2 mL of the internalstandard solution, proceed in the same manner as Primi-done, and use this solution as the standard solution. Per-form the test with 2 mL of the sample solution and the stan-dard solution as directed under the Gas Chromatography ac-cording to the following conditions, and calculate the ratios,QT and QS, of the peak area of 2-ethyl-2-phenylmalonedi-amide to that of the internal standard: QT is not more thanQS.Internal standard solution—A solution of stearylalcohol inpyridine (1 in 2000).Operating conditions—

Detector: A hydrogen ‰ame-ionization detector.Column: A glass column 3 mm in inside diameter and 150

cm in length, packed with siliceous earth for gas chro-matography (125 to 150 mm in particle diameter) coated with50z phenyl-methyl silicon polymer for gas chromatographyat the ratio of 3z.



of stearylalcohol is about 10 minutes.System suitability—

System performance: When the procedure is run with 2 mLof the standard solution under the above operating condi-tion, 2-ethyl-2-phenylmalonediamide and the internal stan-dard are eluted in this order with the resolution betweenthese peaks being not less than 3.

System repeatability: When the test is repeated 5 timeswith 2 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratiosof the peak area of 2-ethyl-2-phenylmalonediamide to thatof the internal standard is not more than 1.5z.

Procaine Hydrochloride Injection塩酸プロカイン注射液


Assay To an exactly measured volume of ProcaineHydrochloride Injection, equivalent to about 20 mg ofprocaine hydrochloride (C13H20N2O2.HCl), add the mobilephase to make exactly 20 mL. Pipet 5 mL of this solution,add exactly 5 mL of the internal standard solution and themobile phase to make 20 mL, and use this solution as thesample solution. Separately, weigh accurately about 50 mgof procaine hydrochloride for assay, previously dried in adesiccator (silica gel) for 4 hours, dissolve in the mobilephase to make exactly 50 mL. Pipet 5 mL of this solution,add exactly 5 mL of the internal standard solution and themobile phase to make 20 mL, and use this solution as thestandard solution. Perform the test with 5 mL each of thesample solution and the standard solution as directed under


the Liquid Chromatography according to the following con-ditions, and calculate the ratios, QT and QS, of the peak areaof procaine to that of the internal standard.

Amount (mg) of procaine hydrochloride (C13H20N2O2.HCl)

＝WS×QT

QS

WS: Amount (mg) of procaine hydrochloride for assay

Internal standard solution—A solution of caŠeine in the mo-bile phase (1 in 1000).Operating conditions—




Mobile phase: Adjust the pH of 0.05 mol/L potassium di-hydrogen phosphate TS to 3.0 with phosphoric acid, andadd an amount of sodium 1-pentane sulfonate to make a so-lution so that containing 0.1z. To 800 mL of this solutionadd 200 mL of methanol.

Flow rate: Adjust the ‰ow rate so that the retention timeof procaine is about 10 minutes.System suitability—

System performance: When the procedure is run with 5 mLof the standard solution under the above operating condi-tions, procaine and the internal standard are eluted in thisorder with the resolution between these peaks being not lessthan 8.

System repeatability: When the test is repeated 6 timeswith 5 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratiosof the peak area of procaine to that of the internal standardis not more than 1.0z.

Procarbazine Hydrochloride塩酸プロカルバジン


Procarbazine Hydrochloride, when dried, containsnot less than 98.5z and not more than 101.0z ofC12H19N3O.HCl.


Description Procarbazine Hydrochloride occurs as whiteto light yellowish white crystals or crystalline powder.


It dissolves in dilute hydrochloric acid.Melting point: about 2239C (with decomposition).


Purity (1) Heavy metals—Proceed with 1.0 g of Procar-bazine Hydrochloride according to Method 4, and performthe test. Prepare the control solution with 2.0 mL of Stan-dard Lead Solution (not more than 20 ppm).

(2) Related substances—Dissolve 50 mg of ProcarbazineHydrochloride in 5.0 mL of a solution of L-cysteinehydrochloride in diluted methanol (7 in 10) (1 in 200), anduse this solution as the sample solution. Pipet 1 mL of thesample solution, add a solution of L-cysteine hydrochloridein diluted methanol (7 in 10) (1 in 200) to make exactly 50mL, and use this solution as the standard solution. Performthe test with these solutions as directed under the Thin-layerChromatography. Immerse slowly, by inclining, a plate ofsilica gel with ‰uorescent indicator for thin-layer chro-matography in a solution of L-cysteine hydrochloride indiluted methanol (7 in 10) (1 in 200), allow to stand for 1minute, lift the plate from the solution, dry it in cold windfor 10 minutes, then dry in warm wind for 5 minutes, andthen dry at 609C for 5 minutes. After cooling, spot 5 mLeach of the sample solution and the standard solution on theplate. Develop the plate with a mixture of methanol andethyl acetate (1:1) to a distance of about 12 cm, and air-drythe plate. Examine under ultraviolet light (main wavelength:254 nm): not more than 1 spot other than the principal spotand the spot of the starting point from the sample solutionappears, and is not more intense than the spot from the stan-dard solution.

Pyrazinamideピラジナミド


Pyrazinamide, when dried, contains not less than99.0z and not more than 101.0z of C5H5N3O.


Description Pyrazinamide occurs as white crystals or crys-talline powder.

It is sparingly soluble in water and in methanol, andslightly soluble in ethanol (99.5).

The pH of a solution obtained by dissolving 1.0 g ofPyrazinamide in 100 mL of water is between 5.0 and 7.0.


Identiˆcation (1) Determine the absorption spectrum ofa solution of Pyrazinamide in 0.1 mol/L hydrochloric acidTS (1 in 100,000) as directed under the Ultraviolet-visibleSpectrophotometry, and compare the spectrum with theReference Spectrum: both spectra exhibit similar intensitiesof absorption at the same wavelengths.

(2) Determine the infrared absorption spectrum ofPyrazinamide, previously dried, as directed in the potassium


bromide disk method under the Infrared Spectrophotomet-ry, and compare the spectrum with the Reference Spectrum:both spectra exhibit similar intensities of absorption at thesame wave numbers.

Change the Purity (4) and (5) to read:

Purity(4) Related substances—Dissolve 0.10 g of Pyrazina-

mide in 10 mL of methanol, and use this solution as the sam-ple solution. Pipet 1 mL of the sample solution, addmethanol to make exactly 200 mL, and use this solution asthe standard solution. Perform the test with these solutionsas directed under the Thin-layer Chromatography. Spot 20mL each of the sample solution and the standard solution ona plate of silica gel with ‰uorescent indicator for thin-layerchromatography. Develop the plate with a mixture of 1-butanol, water and acetic acid (100) (3:1:1) to a distance ofabout 10 cm, and air-dry the plate. Examine under ultravio-let light (main wavelength: 254 nm): the spot other than theprincipal spot obtained from the sample solution is not moreintense than the spot from the standard solution.

(5) Readily carbonizable substances—Perform the testwith 0.20 g of Pyrazinamide: the solution has no more colorthan Matching Fluid A.

Add the following:

Pyrrolnitrinピロールニトリン

C10H6Cl2N2O2: 257.073-Chloro-4-(3-chloro-2-nitrophenyl)pyrrole [1018-71-9 ]

Pyrrolnitrin contains not less than 970 mg (potency)per mg, calculated on the dried basis. The potency ofPyrrolnitrin is expressed as mass (potency) of pyrrol-nitrin (C10H6Cl2N2O2).

Description Pyrrolnitrin occurs as yellow to yellow-brown,crystals or crystalline powder.

It is freely soluble in methanol and in ethanol (95), andpractically insoluble in water.

Identiˆcation (1) Determine the absorption spectrum ofa solution of Pyrrolnitrin in ethanol (95) (1 in 100,000) asdirected under the Ultraviolet-visible Spectrophotometry,and compare the spectrum with the Reference Spectrum orthe spectrum of a solution of Pyrrolnitrin Reference Stan-dard prepared in the same manner as the sample solution:both spectra exhibit similar intensities of absorption at thesame wavelengths.

(2) Determine the infrared absorption spectrum of Pyr-rolnitrin as directed in the potassium bromide disk methodunder the Infrared Spectrophotometry, and compare thespectrum with the Reference Spectrum or the spectrum ofPyrrolnitrin Reference Standard: both spectra exhibit simi-lar intensities of absorption at the same wave numbers.


Purity Related substances—Dissolve 0.10 g of Pyrrolnitrinin 10 mL of methanol, and use this solution as the sample so-lution. Pipet 1 mL of the sample solution, and add methanolto make exactly 100 mL. Pipet 3 mL of this solution, addmethanol to make exactly 10 mL, and use this solution as thestandard solution. Perform the test with these solutions asdirected under the Thin-layer Chromatography. Spot 10 mLeach of the sample solution and the standard solution on aplate of silica gel for thin-layer chromatography. Developthe plate with a mixture of xylene, ethyl acetate and formicacid (18:2:1) to a distance of about 10 cm, and dry the plateat 809C for 30 minutes. Spray evenly diluted sulfuric acid (1in 3) on the plate, and heat at 1009C for 30 minutes: the spotother than the principal spot obtained from the sample solu-tion is not more intense than the spot from the standardsolution.



Assay Conduct this procedure using light-resistant vessels.Weigh accurately an amount of Pyrrolnitrin and Pyrrol-nitrin Reference Standard, equivalent to about 50 mg(potency), dissolve in diluted acetonitrile (3 in 5) to make ex-actly 50 mL. Pipet 10 mL each of these solutions, add ex-actly 10 mL of the internal standard solution, add dilutedacetonitrile (3 in 5) to make 100 mL, and use these solutionsas the sample solution and the standard solution. Performthe test with 5 mL each of the sample solution and the stan-dard solution as directed under the Liquid Chromatographyaccording to the following conditions, and determine the ra-tios, QT and QS, of the peak area of pyrrolnitrin to that ofthe internal standard.

Amount [mg (potency)] of C10H6Cl2N2O2

＝WS×QT

QS× 1000

WS: Amount [mg (potency)] of Pyrrolnitrin ReferenceStandard

Internal standard solution—A solution of benzyl benzoatein diluted acetonitrile (3 in 5) (3 in 500).Operating conditions—






of pyrrolnitrin is about 9 minutes.System suitability—

System performance: When the procedure is run with 5 mLof the standard solution under the above operating condi-tions, pyrrolnitrin and the internal standard are eluted in thisorder with the resolution between these peaks being not lessthan 3.

System repeatability: When the test is repeated 6 timeswith 5 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratiosof the peak area of pyrrolnitrin to that of the internal stan-dard is not more than 1.0z.


Add the following:

Ranitidine Hydrochloride塩酸ラニチジン

C13H22N4O3S.HCl: 350.86N-｛2-[(｛5-[(Dimethylamino)methyl]furan-2-yl｝methyl)sulfanyl]ethyl｝-N?-methyl-2-nitroethene-1,1-diamine monohydrochloride [66357-59-3 ]

Ranitidine Hydrochloride, when dried, contains notless than 97.5z and not more than 102.0z of raniti-dine hydrochloride (C13H22N4O3S.HCl).

Description Ranitidine Hydrochloride occurs as a white topale yellow, crystalline or ˆne granular powder.

It is very soluble in water, freely soluble in methanol, andslightly soluble in ethanol (99.5).

It is hygroscopic.It is gradually colored by light.Melting point: about 1409C (with decomposition).

Identiˆcation (1) Determine the absorption spectrum ofa solution of Ranitidine Hydrochloride (1 in 100,000) asdirected under the Ultraviolet-visible Spectrophotometry,and compare the spectrum with the Reference Spectrum orthe spectrum of a solution of Ranitidine HydrochlorideReference Standard prepared in the same manner as thesample solution: both spectra exhibit similar intensities ofabsorption at the same wavelengths.

(2) Determine the infrared absorption spectrum ofRanitidine Hydrochloride as directed in the paste methodunder the Infrared Spectrophotometry, and compare thespectrum with the Reference Spectrum or the spectrum ofpreviously dried Ranitidine Hydrochloride Reference Stan-dard: both spectra exhibit similar intensities of absorption at

the same wave numbers.(3) A solution of Ranitidine Hydrochloride (1 in 50)

responds to the Qualitative Tests for chloride.

pH The pH of a solution obtained by dissolving 1.0 g ofRanitidine Hydrochloride in 100 mL of water is between 4.5and 6.0.

Purity (1) Clarity and color of solution—A solution ofRanitidine Hydrochloride (1 in 10) is clear and pale yellow tolight yellow.

(2) Heavy metals—Proceed with 2.0 g of RanitidineHydrochloride according to Method 2, and perform the test.Prepare the control solution with 2.0 mL of Standard LeadSolution (not more than 10 ppm).

(3) Arsenic—Prepare the test solution with 1.0 g ofRanitidine Hydrochloride according to Method 4, and per-form the test (not more than 2 ppm).

(4) Related substances—Conduct this procedure withoutexposure to light, using light-resistant vessels. Dissolve 0.22g of Ranitidine Hydrochloride in methanol to make exactly10 mL, and use this solution as the sample solution. Pipet0.5 mL of the sample solution, add methanol to make ex-actly 100 mL, and use this solution as the standard solution(1). Pipet 6 mL, 4 mL, 2 mL and 1 mL of the standard solu-tion (1), add to each methanol to make exactly 10 mL, anduse these solutions as the standard solution (2), the standardsolution (3), the standard solution (4) and the standard solu-tion (5), respectively. Separately, dissolve 12.7 mg ofranitidinediamine in methanol to make exactly 10 mL, anduse this solution as the standard solution (6). Perform thetest with these solutions as directed under the Thin-layerChromatography. Spot 10 mL each of the sample solutionand the standard solutions (1), (2), (3), (4) and (5) on a plateof silica gel for thin-layer chromatography. Separately, spot10 mL of the sample solution on the plate, then spot 10 mL ofthe standard solution (6) on the spotted position of the sam-ple solution. Immediately develop the plate with a mixtureof ethyl acetate, 2-propanol, ammonia solution (28) andwater (25:15:5:1) to a distance of about 15 cm, and air-drythe plate. Allow the plate to stand in iodine vapor until thespot from the standard solution (5) appears: the spot ob-tained from the standard solution (6) is completely separatedfrom the principal spot from the sample solution. The spothaving Rf value of about 0.7 from the sample solution is notmore intense than the spot from the standard solution (1),the spots other than the principal spot and the spot of Rf 0.7from the sample solution are not more intense than the spotfrom the standard solution (2), and the total amount of theserelated substances, calculated by comparison with the spotsfrom the standard solutions (1), (2), (3), (4) and (5), is notmore than 1.0z.



Assay Weigh accurately about 20 mg of RanitidineHydrochloride and Ranitidine Hydrochloride Reference


Standard, previously dried, dissolve each in the mobilephase to make exactly 200 mL, and use these solutions as thesample solution and the standard solution. Perform the testwith exactly 10 mL each of the sample solution and the stan-dard solution as directed under the Liquid Chromatographyaccording to the following conditions, and determine thepeak areas, AT and AS, of ranitidine.

Amount (mg) of C13H22N4O3S.HCl＝WS×AT

AS

WS: Amount (mg) of Ranitidine Hydrochloride ReferenceStandard





Mobile phase: A mixture of methanol and diluted 0.5mol/L ammonium acetate TS (1 in 5) (17:3).

Flow rate: Adjust the ‰ow rate so that the retention timeof ranitidine is about 5 minutes.System suitability—

System performance: Dissolve 20 mg of RanitidineHydrochloride and 5 mg of benzalphthalide in 200 mL ofthe mobile phase. When the procedure is run with 10 mL ofthis solution under the above operating conditions, ben-zalphthalide and ranitidine are eluted in this order with theresolution between these peaks being not less than 2.0.

System repeatability: When the test is repeated 6 timeswith 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peakarea of ranitidine is not more than 1.0z.


Reserpineレセルピン


Purity Related substances—Conduct this procedurewithout exposure to daylight, using light-resistant vessels.Dissolve 50 mg of Reserpine in 50 mL of acetonitrile, anduse this solution as the sample solution. Pipet 3 mL of thesample solution, add acetonitrile to make exactly 100 mL,and use this solution as the standard solution. Perform thetest with exactly 10 mL each of the sample solution and thestandard solution as directed under the Liquid Chro-matography according to the following conditions. Deter-mine each peak area from these solutions by the automaticintegration method: the total area of all peaks other than

reserpine peak from the sample solution is not larger thanthe peak area of reserpine from the standard solution.Operating conditions—

Detector, column, and column temperature: Proceed asdirected in the operating conditions in the Assay.

Mobile phase: A mixture of 0.05 mol/L potassium di-hydrogen phosphate, pH 3.0 and acetonitrile (13:7).

Flow rate: Adjust the ‰ow rate so that the retention timeof reserpine is about 20 minutes.

Time span of measurement: About twice as long as theretention time of reserpine.System suitability—

Test for required detection: To exactly 2 mL of the stan-dard solution add acetonitorile to make exactly 50 mL. Con-ˆrm that the peak area of reserpine obtained from 10 mL ofthis solution is equivalent to 3 to 5z of that of reserpine ob-tained from 10 mL of the standard solution.

System performance: Dissolve 0.01 g of Reserpine and 4mg of butyl parahydroxybenzoate in 100 mL of acetonitrile.To 5 mL of this solution add acetonitrile to make 50 mL.When the procedure is run with 20 mL of this solution ac-cording to the operating conditions in the Assay, reserpineand butyl parahydroxybenzoate are eluted in this order withthe resolution between these peaks being not less than 2.0.

System repeatability: When the test is repeated 6 timeswith 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peakarea of reserpine is not more than 2.0z.


Assay Conduct this procedure without exposure to day-light, using light-resistant vessels. Weigh accurately about 10mg each of Reserpine and Reserpine Reference Standard,previously dried, and dissolve each in acetonitrile to makeexactly 100 mL. Pipet 5 mL each of these solutions, add ex-actly 10 mL of the internal standard solution, 5 mL ofacetonitrile and water to make 50 mL, and use these solu-tions as the sample solution and the standard solution, re-spectively. Perform the test with 20 mL each of the samplesolution and the standard solution as directed under the Liq-uid Chromatography according to the following conditions,and calculate the ratios, QT and QS, of the peak area ofreserpine to that of the internal standard.

Amount (mg) of C33H40N2O9＝WS×QT

QS

WS: Amount (mg) of Reserpine Reference Standard

Internal standard solution—A solution of butyl parahydrox-ybenzoate in acetonitrile (1 in 50,000).Operating conditions—





Mobile phase: A mixture of 0.05 mol/L potassium di-hydrogen phosphate, pH 3.0 and acetonitrile (11:9).

Flow rate: Adjust the ‰ow rate so that the retention timeof reserpine is about 10 minutes.System suitability—

System performance: When the procedure is run with 20mL of the standard solution under the above operating con-ditions, reserpine and the internal standard are eluted in thisorder with the resolution between these peaks being not lessthan 2.0.

System repeatability: When the test is repeated 6 timeswith 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratiosof the peak area of reserpine to that of the internal standardis not more than 2.0z.

Retinol Acetate酢酸レチノール


Retinol Acetate is synthetic retinol acetate or syn-thetic retinol acetate diluted with ˆxed oil. It containsnot less than 2,500,000 Vitamin A Units per gram. Asuitable antioxidant may be added.

Retinol Acetate contains not less than 95.0z andnot more than 105.0z of the labeled Units.

Description Retinol Acetate occurs as pale yellow to yel-low-red crystals or an ointment-like substance, and has afaint, characteristic odor, but has no rancid odor.

It is freely soluble in petroleum ether, soluble in ethanol(95), and practically insoluble in water.

It is decomposed by air and by light.

Identiˆcation Dissolve Retinol Acetate and RetinolAcetate Reference Standard, equivalent to 15,000 Units, in 5mL of petroleum ether, and use these solutions as the samplesolution and the standard solution. Perform the test withthese solutions as directed under the Thin-layer Chro-matography. Spot 5 mL each of the sample solution and thestandard solution on a plate of silica gel for thin-layer chro-matography. Develop with a mixture of cyclohexane anddiethyl ether (12:1) to a distance of about 10 cm, and air-drythe plate. Spray evenly antimony (III) chloride TS: the prin-cipal spot obtained from the sample solution is the same incolor tone and Rf value with the blue spot from the standardsolution.

Purity (1) Acid value—Take exactly 5.0 g of RetinolAcetate, and perform the test: not more than 2.0.

(2) Peroxide—Weigh accurately about 5 g of RetinolAcetate, transfer in a 250-mL glass-stoppered conical ‰ask,add 50 mL of a mixture of acetic acid (100) and isooctane(3:2), and gently mix to dissolve completely. Replace the airof the inside gradually with about 600 mL of Nitrogen, then

add 0.1 mL of saturated potassium iodide TS under a cur-rent of Nitrogen. Immediately stopper tightly, and mix witha swirling motion for 1 minute. Add 30 mL of water, stop-per tightly, and shake vigorously for 5 to 10 seconds. Titratethis solution with 0.01 mol/L sodium thiosulfate VS untilthe blue color of the solution disappears after addition of 0.5mL of starch TS near the end point where the solution is apale yellow color. Calculate the amount of peroxide by thefollowing formula: not more than 10 meq/kg.

Amount (meq/kg) of peroxide＝VW× 10

V: Volume (mL) of 0.01 mol/L sodium thiosulfate VSconsumed

W: Amount (g) of the sample

Assay Proceed as directed in Method 1-1 under the Vita-min A Assay.

Containers and storage Containers—Tight containers.Storage—Light-resistant, and almost well-ˆlled, or under

Nitrogen atmosphere, and in a cold place.

Retinol Palmitateパルミチン酸レチノール


Retinol Palmitate is a synthetic retinol palmitate ora synthetic retinol palmitate diluted with ˆxed oil, andcontains not less than 1,500,000 Vitamin A Units ineach gram. It may contain a suitable antioxidant.

Retinol Palmitate contains not less than 95.0z andnot more than 105.0z of the labeled Units.

Description Retinol Palmitate occurs as a light yellow toyellow-red, ointment-like or an oily substance. It has a faint,characteristic odor, but has no rancid odor.

It is very soluble in petroleum ether, slightly soluble inethanol (95), and practically insoluble in water.

It is decomposed by air and by light.

Identiˆcation Dissolve Retinol Palmitate and RetinolPalmitate Reference Standard, equivalent to 15,000 Units,in 5 mL of petroleum ether, and use these solutions as thesample solution and the standard solution. Perform the testwith these solutions as directed under the Thin-layer Chro-matography. Spot 5 mL each of the sample solution and thestandard solution on a plate of silica gel for thin-layer chro-matography. Develop with a mixture of cyclohexane anddiethyl ether (12:1) to a distance of about 10 cm, and air-drythe plate. Spray evenly antimony (III) chloride TS: the prin-cipal spot obtained from the sample solution is the same incolor tone and Rf value with the blue spot from the standardsolution.

Purity (1) Acid value—Take exactly 5.0 g of Retinol


Palmitate, and perform the test: not more than 2.0.(2) Peroxide—Weigh accurately about 5 g of Retinol

Palmitate, transfer in a 250-mL glass-stoppered conical‰ask, add 50 mL of a mixture of acetic acid (100) and isooc-tane (3:2), and gently mix to dissolve completely. Replacethe air of the inside gradually with about 600 mL of Nitro-gen, then add 0.1 mL of saturated potassium iodide TSunder a current of Nitrogen. Immediately stopper tightly,and mix with a swirling motion for 1 minute. Add 30 mL ofwater, stopper tightly, and shake vigorously for 5 to 10seconds. Titrate this solution with 0.01 mol/L sodiumthiosulfate VS until the blue color of the solution disappearsafter addition of 0.5 mL of starch TS near the end pointwhere the solution is a pale yellow color. Calculate theamount of peroxide by the following formula: not morethan 10 meq/kg.

Amount (meq/kg) of peroxide＝VW× 10

V: Volume (mL) of 0.01 mol/L sodium thiosulfate VSW: Amount (g) of the sample



Nitrogen atmosphere, and in a cold place.

Ribostamycin Sulfate硫酸リボスタマイシン


Ribostamycin Sulfate contains not less than 680 mg(potency) per mg, calculated on the dried basis. Thepotency of Ribostamycin Sulfate is expressed as mass(potency) of ribostamycin (C17H34N4O10: 454.47).

Description Ribostamycin Sulfate occurs as a white to yel-lowish white powder.


Identiˆcation (1) Dissolve 20 mg of Ribostamycin Sul-fate in 2 mL of phosphate buŠer solution, pH 6.0, add 1 mLof ninhydrin TS, and boil: a blue-purple color develops.

(2) Dissolve 0.12 g each of Ribostamycin Sulfate andRibostamycin Sulfate Reference Standard in 20 mL ofwater, and use these solutions as the sample solution and thestandard solution. Perform the test with these solutions asdirected under the Thin-layer Chromatography. Spot 5 mLeach of the sample solution and the standard solution on aplate of silica gel for thin-layer chromatography. Developthe plate with a solution of potassium dihydrogen phosphate(3 in 40) to a distance of about 10 cm, and air-dry the plate.Spray evenly 0.2z ninhydrin-water saturated 1-butanol TS,

and heat at 1009C for 10 minutes: the principal spots ob-tained from the sample solution and the standard solutionshow a purple-brown color and the same Rf value.

(3) To 2 mL of a solution of Ribostamycin Sulfate (1 in5) add 1 drop of barium chloride TS: a white turbidity is pro-duced.

Optical rotation [a]D20: ＋42 –＋499(after drying, 0.25 g,water, 25 mL, 100 mm).

pH The pH of a solution obtained by dissolving 1.0 g ofRibostamycin Sulfate in 20 mL of water is between 6.0 and8.0.

Purity (1) Clarity and color of solution—Dissolve 1.0 gof Ribostamycin Sulfate in 5 mL of water: the solution isclear, and colorless or pale yellow.

(2) Heavy metals—Proceed with 1.0 g of RibostamycinSulfate according to Method 1, and perform the test. Pre-pare the control solution with 3.0 mL of Standard Lead So-lution (not more than 30 ppm).

(3) Arsenic—Prepare the test solution with 1.0 g ofRibostamycin Sulfate according to Method 1, and performthe test (not more than 2 ppm).

(4) Related substances—Dissolve 0.12 g of Ribostamy-cin Sulfate in water to make exactly 20 mL, and use this so-lution as the sample solution. Pipet 5 mL of the sample solu-tion, add water to make exactly 100 mL, and use this solu-tion as the standard solution. Perform the test with these so-lutions as directed under the Thin-layer Chromatography.Spot 5 mL each of the sample solution and the standard solu-tion on a plate of silica gel for thin-layer chromatography.Develop the plate with a solution of potassium dihydrogenphosphate (3 in 40) to a distance of about 10 cm, and air-drythe plate. Spray evenly 0.2z ninhydrin-water saturated 1-butanol TS on the plate, and heat at 1009C for 10 minutes:the spot other than the principal spot obtained from thesample solution is not more intense than the spot from thestandard solution.






(3) Standard solutions—Weigh accurately an amount ofRibostamycin Sulfate Reference Standard, previously dried,equivalent to about 20 mg (potency), dissolve in dilutedphosphate buŠer solution, pH 6.0 (1 in 2) to make exactly 50mL, and use this solution as the standard stock solution.Keep the standard stock solution at 5 to 159C and use within20 days. Take exactly a suitable amount of the standardstock solution before use, add 0.1 mol/L phosphate buŠersolution, pH 8.0 to make solutions so that each mL contains


20 mg (potency) and 5 mg (potency), and use these solutionsas the high concentration standard solution and the low con-centration standard solution, respectively.

(4) Sample solutions—Weigh accurately an amount ofRibostamycin Sulfate, equivalent to about 20 mg (potency),and dissolve in water to make exactly 50 mL. Take exactly asuitable amount of this solution, add 0.1 mol/L phosphatebuŠer solution, pH 8.0 to make solutions so that each mLcontains 20 mg (potency) and 5 mg (potency), and use thesesolutions as the high concentration sample solution and thelow concentration sample solution, respectively.


Rifampicinリファンピシン


Rifampicin contains not less than 970 mg (potency)and not more than 1020 mg (potency) per mg, calculat-ed on the dried basis. The potency of Rifampicin is ex-pressed as mass (potency) of rifampicin (C43H58N4O12:822.94).

Description Rifampicin occurs as orange-red to red-brown, crystals or crystalline powder.

It is slightly soluble in water, in acetonitrile, in methanoland in ethanol (95).

Identiˆcation (1) To 5 mL of a solution of Rifampicin inmethanol (1 in 5000) add 0.05 mol/L phosphate buŠer solu-tion, pH 7.0 to make 100 mL. Determine the absorptionspectrum of this solution as directed under the Ultraviolet-visible Spectrophotometry, and compare the spectrum withthe Reference Spectrum or the spectrum of a solution ofRifampicin Reference Standard prepared in the same man-ner as the sample solution: both spectra exhibit similar inten-sities of absorption at the same wavelengths.

(2) Determine the infrared absorption spectrum ofRifampicin as directed in the potassium bromide diskmethod under the Infrared Spectrophotometry, and com-pare the spectrum with the Reference Spectrum or the spec-trum of Rifampicin Reference Standard: both spectraexhibit similar intensities of absorption at the same wavenumbers.

Purity (1) Heavy metals—Proceed with 1.0 g of Rifampi-cin according to Method 2, and perform the test. Prepare thecontrol solution with 2.0 mL of Standard Lead Solution (notmore than 20 ppm).

(2) Arsenic—Prepare the test solution with 1.0 g ofRifampicin according to Method 3, and perform the test(not more than 2 ppm).

(3) Related substances—Perform the test immediatelyafter preparing of the sample and standard solutions. Dis-solve 0.10 g of Rifampicin in 50 mL of acetonitrile, and use

this solution as the sample stock solution. Pipet 5 mL of thesample stock solution, add citric acid-phosphate-acetonitrileTS to make exactly 50 mL, and use this solution as the sam-ple solution. Separately, pipet 1 mL of the sample stock so-lution, and add acetonitrile to make exactly 100 mL. Pipet 5mL of this solution, add citric acid-phosphate-acetonitrileTS to make exactly 50 mL, and use this solution as the stan-dard solution. Perform the test with exactly 50 mL each ofthe sample solution and the standard solution as directed un-der the Liquid Chromatography according to the followingconditions, and determine each peak area by the automaticintegration method: the area of the peak appeared at the rel-ative retention time of about 0.7 with respect to rifampicinfrom the sample solution is not more than 1.5 times the peakarea of rifampicin from the standard solution, the area ofthe peak other than rifampicin and the peak mentionedabove from the sample solution is not more than the peakarea of rifampicin from the standard solution, and the totalarea of the peaks other than rifampicin and the peak men-tioned above from the sample solution is not more than 3.5times the peak area of rifampicin from the standard solu-tion.Operating conditions—


Time span of measurement: About 3 times as long as theretention time of rifampicin after the peak of the solvent.System suitability—

Test for required detectability: Measure exactly 2 mL ofthe standard solution, and add citric acid-phosphate-acetonitrile TS to make exactly 20 mL. Conˆrm that thepeak area of rifampicin obtained from 50 mL of this solutionis equivalent to 7 to 13z of that from 50 mL of the standardsolution.


System repeatability: When the test is repeated 6 timeswith 50 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peakarea of rifampicin is not more than 2.0z.



Assay Weigh accurately an amount of Rifampicin andRifampicin Reference Standard, equivalent to about 40 mg(potency), and dissolve each in acetonitrile to make exactly200 mL. Pipet 10 mL each of these solutions, add citric acid-phosphate-acetonitrile TS to make exactly 100 mL, and usethese solutions as the sample solution and the standard solu-tion. Perform the test with 50 mL each of the sample solutionand the standard solution as directed under the Liquid Chro-matography according to the following conditions, and de-termine the peak areas, AT and AS, of rifampicin.



＝WS×AT

AS× 1000

WS: Amount [mg (potency)] of Rifampicin ReferenceStandard



ameter and 10 cm in length, packed with octylsilanized silicagel for liquid chromatography (5 mm in particle diameter).


Mobile phase: Dissolve 4.2 g of citric acid monohydrateand 1.4 g of sodium perchlorate in 1000 mL of a mixture ofwater, acetonitrile and phosphate buŠer solution, pH 3.1(11:7:2).

Flow rate: Adjust the ‰ow rate so that the retention timeof rifampicin is about 8 minutes.System suitability—

System performance: To 5 mL of a solution of Rifampicinin acetonitrile (1 in 5000) add 1 mL of a solution of butylparahydroxybenzoate in acetonitrile (1 in 5000) and citricacid-phosphate-acetonitrile TS to make 50 mL. When theprocedure is run with 50 mL of this solution under the aboveoperating conditions, butyl parahydroxybenzoate andrifampicin are eluted in this order with the resolution be-tween these peaks being not less than 1.5.

System repeatability: When the test is repeated 5 timeswith 50 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peakarea of rifampicin is not more than 1.0z.


Roxithromycinロキシスロマイシン


Description Roxithromycin occurs as a white crystallinepowder.

It is freely soluble in ethanol (95) and in acetone, solublein methanol, sparingly soluble in acetonitrile, and practicallyinsoluble in water.

Scopolamine Butylbromide臭化ブチルスコポラミン


Purity(3) Related substances—Dissolve 0.10 g of Scopolamine

Butylbromide in the mobile phase to make exactly 10 mL,and use this solution as the sample solution. Separately, dis-solve 10 mg of scopolamine hydrobromide in the mobilephase to make exactly 100 mL. Pipet 10 mL of this solution,add the mobile phase to make exactly 50 mL, and use this so-lution as the standard solution (1). Pipet 5 mL of the stan-dard solution (1), add the mobile phase to make exactly 10mL, and use this solution as the standard solution (2). Per-form the test with exactly 20 mL each of the sample solutionand the standard solutions (1) and (2) as directed under theLiquid Chromatography according to the following condi-tions. Determine each peak area of these solutions by the au-tomatic integration method: the peak area of scopolaminefrom the sample solution is not larger than that from thestandard solution (2), and each area of the peaks other thanthe peak appearing in the ˆrst elution and the peak ofscopolamine and butylscopolamine from the sample solu-tion are not larger than the peak area from the standard so-lution (1).Operating conditions—


Column: A stainless steel column 4.6 mm in inside di-ameter and 15 cm in length, packed with octylsilanized silicagel for liquid chromatography (10 mm in particle diameter).


Mobile phase: Dissolve 2 g of sodium lauryl sulfate in 370mL of water and 680 mL of methanol, and adjust the pH to3.6 with diluted phosphoric acid (1 in 10).

Flow rate: Adjust the ‰ow rate so that the retention timeof butylscopolamine is about 7 minutes.

Time span of measurement: About twice as long as theretention time of butylscopolamine.System suitability—

System performance: Dissolve 5 mg each of ScopolamineButylbromide and scopolamine hydrobromide in 50 mL ofthe mobile phase. When the procedure is run with 20 mL ofthis solution under the above operating conditions, scopola-mine and butylscopolamine are eluted in this order with theresolution between these peaks being not less than 5.

System repeatability: When the test is repeated 6 timeswith 20 mL of the standard solution (2) under the aboveoperating conditions, the relative standard deviation of thepeak area of scopolamine is not more than 2.0z.


Add the following:

Siccaninシッカニン

C22H30O3: 342.47(4aS,6aS,11bR,13aS,13bS )-1,2,3,4,4a,5,6,6a,11b,13b-Decahydro-4,4,6a,9-tetramethyl-13H-benzo[a]furo[2,3,4-mn]xanthen-11-ol [22733-60-4 ]

Siccanin contains not less than 980 mg (potency) permg, calculated on the dried basis. The potency ofSiccanin is expressed as mass (potency) of siccanin(C22H30O3).

Description Siccanin occurs as white to light yellow, crys-tals or crystalline powder.

It is freely soluble in acetone, soluble in methanol and inethanol (99.5), and practically insoluble in water.

Identiˆcation (1) Determine the absorption spectrum ofa solution of Siccanin in ethanol (99.5) (1 in 10,000) asdirected under the Ultraviolet-visible Spectrophotometry,and compare the spectrum with the Reference Spectrum orthe spectrum of a solution of Siccanin Reference Standardobtained in the same manner as the sample solution: bothspectra exhibit similar intensities of absorption at the samewavelengths.

(2) Determine the infrared absorption spectrum of Sic-canin as directed in the potassium bromide disk method un-der the Infrared Spectrophotometry, and compare the spec-trum with the Reference Spectrum or the spectrum of Sicca-nin Reference Standard: both spectra exhibit similar intensi-ties of absorption at the same wave numbers.

Optical rotation [a]D20: －165 –－1759 (0.1 g, ethanol(99.5), 10 mL, 100 mm).


Purity (1) Heavy metals—Proceed with 1.0 g of Siccaninaccording to Method 4, and perform the test. Prepare thecontrol solution with 2.0 mL of Standard Lead Solution (notmore than 20 ppm).

(2) Related substances—Dissolve 0.20 g of Siccanin in10 mL of acetone, and use this solution as the sample solu-tion. Pipet 1 mL of the sample solution, add acetone tomake exactly 200 mL, and use this solution as the standardsolution. Perform the test with these solutions as directedunder the Thin-layer Chromatography. Spot 5 mL each ofthe sample solution and the standard solution on a plate ofsilica gel for thin-layer chromatography. Develop the platewith a mixture of cyclohexane and acetone (5:1) to a distanceof about 10 cm, and air-dry the plate. Spray evenly 4-chlo-robenzenediazonium TS on the plate: the number of the

spots other than the principal spot obtained from the samplesolution is not more than three, and they are not more in-tense than the spot from the standard solution.



Assay Weigh accurately an amount of Siccanin and Sicca-nin Reference Standard, equivalent to about 50 mg (poten-cy), dissolve each in the internal standard solution to makeexactly 50 mL, and use these solutions as the sample solutionand the standard solution. Perform the test with 10 mL eachof the sample solution and the standard solution as directedunder the Liquid Chromatography according to the follow-ing conditions, and determine the ratios, QT and QS, of thepeak area of siccanin to that of the internal standard.

Amount [mg (potency)] of C22H30O3＝WS×QT

QS× 1000

WS: Amount [mg (potency)] of Siccanin Reference Stand-ard

Internal standard solution—A solution of 1,4-diphenylben-zene in methanol (1 in 30,000).Operating conditions—




Mobile phase: A mixture of methanol and phosphatebuŠer solution, pH 5.9 (19:6).

Flow rate: Adjust the ‰ow rate so that the retention timeof siccanin is about 17 minutes.System suitability—

System performance: When the procedure is run with 10mL of the standard solution under the above operating con-ditions, siccanin and the internal standard are eluted in thisorder with the resolution between these peaks being not lessthan 3.0.

System repeatability: When the test is repeated 6 timeswith 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratiosof the peak area of siccanin to that of the internal standard isnot more than 1.0z.




Simˆbrateシンフィブラート

Sisomicin Sulfate硫酸シソマイシン


Purity(3) Related substances—Dissolve 50 mg of Sisomicin

Sulfate, calculated on the dried basis, in water to make 10mL, and use this solution as the sample solution. Pipet 0.5mL, 1 mL and 1.5 mL of the sample solution, add water toeach to make exactly 50 mL, and use these solutions as thestandard solution (1), the standard solution (2) and the stan-dard solution (3), respectively. Perform the test with thesesolutions as directed under the Thin-layer Chromatography.Spot 10 mL each of the sample solution and the standard so-lutions (1), (2), and (3) on a plate of silica gel for thin-layerchromatography. Develop the plate with a mixture ofmethanol, chloroform, ammonia water (28) and acetone(2:2:1:1) to a distance of about 10 cm, and air-dry the plate.Spray evenly 0.2z ninhydrin-water saturated 1-butabol TSon the plate, and heat at 1009C for 5 minutes. The spots cor-responding to Rf about 0.35 and Rf about 0.30 are not moreintense than that of the spot from the standard solution (3),and the spot of gallamine corresponding to Rf about 0.25 isnot more intense than the spot from the standard solution(1). The total amount of the related substances is not morethan 6z.

Sodium Bicarbonate Injection炭酸水素ナトリウム注射液


pH 7.0 – 8.5

Delete the Purity.



Change to read:

Sodium Chloride塩化ナトリウム

NaCl: 58.44

Sodium Chloride, when dried, contains not less than99.0z and not more than 100.5z of NaCl.

Description Sodium Chloride occurs as colorless or white,crystals or crystalline powder.


Identiˆcation (1) A solution of Sodium Chloride (1 in 20)responds to the Qualitative Tests for sodium salt.

(2) A solution of Sodium Chloride (1 in 20) responds tothe Qualitative Tests for chloride.

Purity (1) Clarity and color of solution—Dissolve 1.0 gof Sodium Chloride in 5 mL of water: the solution is clearand colorless.

(2) Acidity or alkalinity—Dissolve 20.0 g of SodiumChloride in 100.0 mL of freshly boiled and cooled water,and use this solution as the sample solution. To 20 mL of thesample solution add 0.1 mL of bromothymol blue TS and0.5 mL of 0.01 mol/L hydrochloric acid VS: the color of thesolution is yellow. Separately, to 20 mL of the sample solu-tion add 0.1 mL of bromothymol blue TS and 0.5 mL of0.01 mol/L sodium hydroxide VS: the color of the solutionis blue.

(3) Sulfates—To 7.5 mL of the sample solution obtainedin (2) add 30 mL of water, and use this solution as the sam-ple solution. Separately, dissolve 0.181 g of potassium sul-fate in diluted ethanol (99.5) (3 in 10) to make exactly 500mL. Pipet 5 mL of this solution, and add diluted ethanol(99.5) (3 in 10) to make exactly 100 mL. To 4.5 mL of thissolution add 3 mL of a solution of barium chloride dihy-drate (1 in 4), shake, and allow to stand for 1 minutes. To2.5 mL of this solution add 15 mL of the sample solutionand 0.5 mL of acetic acid (31), and allow to stand for 5minutes: any turbidity produced does not more than thatproduced in the following control solution.

Control solution: Dissolve 0.181 g of potassium sulfate inwater to make exactly 500 mL. Pipet 5 mL of this solution,add water to make exactly 100 mL, and proceed in the samemanner as directed above using this solution instead of thesample solution.

(4) Phosphates—To 2.0 mL of the sample solution ob-tained in (2) add 5 mL of 2 mol/L sulfuric acid TS and waterto make 100.0 mL, then add 4 mL of ammonium molyb-date-sulfuric acid TS and 0.1 mL of tin (II) chloride-hydrochloric acid TS, and allow to stand for 10 minutes: thecolor of the solution is not darker than the following controlsolution.

Control solution: To 1.0 mL of Standard PhosphoricAcid Solution add 12.5 mL of 2 mol/L sulfuric acid TS and


water to make exactly 250 mL. To 100 mL of this solutionadd 4 mL of ammonium molybdate-sulfuric acid TS and 0.1mL of tin (II) chloride-hydrochloric acid TS, and allow tostand for 10 minutes.

(5) Bromides—To 0.50 mL of the sample solution ob-tained in (2) add 4.0 mL of water, 2.0 mL of dilute phenolred TS and 1.0 mL of a solution of sodium toluenesulfon-chloramide trihydrate (1 in 10,000), and mix immediately.After allowing to stand for 2 minutes, add 0.15 mL of 0.1mol/L sodium thiosulfate VS, mix, add water to make ex-actly 10 mL, and use this solution as the sample solution.Separately, to 5.0 mL of a solution of potassium bromide (3in 1,000,000) add 2.0 mL of dilute phenol red TS and 1.0 mLof a solution of sodium toluenesulfonchloramide trihydrate(1 in 10,000), and mix immediately. Proceed in the samemanner as for the preparation of the sample solution, anduse the solution so obtained as the standard solution. Per-form the test with the sample solution and the standard solu-tion as directed under the Ultraviolet-visible Spectrophoto-metry using water as the control: the absorbance at 590 nmof the sample solution is not more than that of the standardsolution.

(6) Iodides—Wet 5 g of Sodium Chloride with drop-wisely added 0.15 mL of a freshly prepared mixture of starchTS, 0.5 mol/L sulfuric acid TS and sodium nitrite TS(1000:40:3), allow to stand for 5 minutes, and examine un-der daylight: a blue color does not appear.

(7) Ferrocyanides—Dissolve 2.0 g of Sodium Chloridein 6 mL of water, and add 0.5 mL of a mixture of a solutionof iron (II) sulfate heptahydrate (1 in 100) and a solution ofammonium iron (III) sulfate 12-water in diluted sulfuric acid(1 in 400) (1 in 100) (19:1): a blue color does not develop wi-thin 10 minutes.

(8) Heavy metals—Proceed with 5.0 g of Sodium Chlo-ride according to Method 1, and perform the test. Preparethe control solution with 1.5 mL of Standard Lead Solution(not more than 3 ppm).

(9) Iron—To 10 mL of the sample solution obtained in(2) add 2 mL of a solution of citric acid monohydrate (1 in 5)and 0.1 mL of mercapto acetic acid, alkalize with ammoniaTS, add water to make 20 mL, and allow to stand for 5minutes: the solution has not more color than the followingcontrol solution.

Control solution: Pipet 1 mL of Standard Iron Solution,and add water to make exactly 25 mL. To 10 mL of this so-lution add 2 mL of a solution of citric acid monohydrate (1in 5) and 0.1 mL of mercapto acetic acid, and proceed in thesame manner as directed for the sample solution.

(10) Barium—To 5.0 mL of the sample solution ob-tained in (2) add 5.0 mL of water and 2.0 mL of dilute sul-furic acid, and allow to stand for 2 hours: the solution hasnot more turbidity than the following control solution.

Control solution: To 5.0 mL of the sample solution ob-tained in (2) add 7.0 mL of water, and allow to stand for 2hours.

(11) Magnesium and alkaline-earth materials—To 200mL of water add 0.1 g of hydroxylammonium chloride, 10mL of ammonium chloride buŠer solution, pH 10, 1 mL of

0.1 mol/L zinc sulfate VS and 0.2 g of eriochrome black T-sodium chloride indicator, and warm to 409C. Add 0.01mol/L disodium dihydrogen ethylenediamine tetraacetateVS dropwise until the red-purple color of the solutionchanges to blue-purple. To this solution add a solution pre-pared by dissolving 10.0 g of Sodium Chloride in 100 mL ofwater, and add 2.5 mL of 0.01 mol/L disodium dihydrogenethylenediamine tetraacetate VS: the color of the solution isa blue-purple.

(12) Arsenic—Prepare the test solution with 1.0 g of So-dium Chloride according to Method 1, and perform the test(not more than 2 ppm).


Assay Weigh accurately about 50 mg of Sodium Chloride,previously dried, dissolve in 50 mL of water, and titrate with0.1 mol/L silver nitrate VS (potentiometric titration).

Each mL of 0.1 mol/L silver nitrate VS＝ 5.844 mg of NaCl


10z Sodium Chloride Injection10 塩化ナトリウム注射液



Add the following:

Sodium Fusidateフシジン酸ナトリウム

C31H47NaO6: 538.69Monosodium (17Z)-ent-16a-acetoxy-3b,11b-dihydroxy-4b,8b,14a-trimethyl-18-nor-5b,10a-cholesta-17(20),24-dien-21-oate [751-94-0 ]

Sodium Fusidate contains not less than 935 mg(potency) per mg, calculated on the anhydrous basis.The potency of Sodium Fusidate is expressed as mass(potency) of fusidic acid (C31H48O6: 516.71).

Description Sodium Fusidate occurs as white, crystals of


crystalline powder.It is freely soluble in water, in methanol and in ethanol

(99.5).

Identiˆcation (1) Determine the infrared absorptionspectra of Sodium Fusidate as directed in the potassiumbromide disk method under the Infrared Spectrophotomet-ry, and compare the spectrum with the Reference Spectrum:both spectra exhibit similar intensities of absorption at thesame wave numbers.

(2) Sodium Fusidate responds to the Qualitative Test (1)for sodium salt.

Purity Heavy metals—Proceed with 2.0 g of Sodium Fusi-date according to Method 4, and perform the test. Preparethe control solution with 2.0 mL of Standard Lead Solution(not more than 10 ppm).




(2) Culture medium—Use the medium ii in 3) Mediumfor other organisms under (1) Agar media for seed and baselayer.

(3) Standard solutions—Weigh accurately an amount ofDiethanolamine Fusidate Reference Standard, equivalent toabout 20 mg (potency), dissolve in 2 mL of ethanol (95), addwater to make exactly 20 mL, and use this solution as thestandard stock solution. Keep the standard stock solution ata temperature not exceeding 59C and use within 7 days.Take exactly a suitable amount of the standard stock solu-tion before use, add phosphate buŠer solution, pH 6.0 tomake solutions so that each mL contains 4 mg (potency) and1 mg (potency), and use these solutions as the high concentra-tion standard solution and the low concentration standardsolution, respectively.

(4) Sample solutions—Weigh accurately an amount ofSodium Fusidate, equivalent to about 20 mg (potency), anddissolve in water to make exactly 20 mL. Take exactly a suit-able amount of this solution, add phosphate buŠer solution,pH 6.0 to make solutions so that each mL contains 4 mg(potency) and 1 mg (potency), and use these solutions as thehigh concentration sample solution and the low concentra-tion sample solution, respectively.

Containers and storage Containers—Tight containers.Storage—Light-resistant, and at a temperature 2 to 89C.


Sodium Iopodateイオポダートナトリウム

Sodium Iopodate Capsulesイオポダートナトリウムカプセル

Add the following:

Spectinomycin Hydrochloride塩酸スペクチノマイシン

C14H24N2O7.2HCl.5H2O: 495.35(2R,4aR,5aR,6S,7S,8R,9S,9aR,10aS )-Decahydro-4a,7,9-trihydroxy-2-methyl-6,8-bis(methylamino)-4H-pyrano[2,3-b][1,4]benzodioxin-4-onedihydrochloride pentahydrate [22189-32-8 ]

Spectinomycin Hydrochloride contains not less than603 mg (potency) per mg. The potency of Spectinomy-cin Hydrochloride is expressed as mass (potency) ofspectinomycin (C14H24N2O7: 332.35).

Description Spectinomycin Hydrochloride occurs as awhite to light yellowish white crystalline powder.


Identiˆcation (1) To 5 mL of a solution of Spectinomy-cin Hydrochloride (1 in 100) add gently anthrone TS: a blueto blue-green color is produced at the zone of contact.

(2) Determine the infrared absorption spectra of Spec-tinomycin Hydrochloride and Spectinomycin HydrochlorideReference Standard as directed in the paste method underthe Infrared Spectrophotometry, and compare these spectra:both spectra exhibit similar intensities of absorption at thesame wave numbers.

(3) To 3 mL of a solution of Spectinomycin Hydrochlo-ride (1 in 150) add 1 drop of silver nitrate TS: a white turbid-ity is produced.

Optical rotation [a]D20: ＋15 –＋219(2.1 g calculated onthe anhydrous basis, water, 25 mL, 200 mm).

pH Dissolve 0.10 g of Spectinomycin Hydrochloride in 10mL of water: the pH of the solution is between 4.0 and 5.6.

Water Not less than 16.0z and not more than 20.0z

(0.3 g, volumetric titration, direct titration).




(1) Test organism—Klebsiella pneumoniae ATCC 10031(2) Culture medium—Use the medium i in 3) Medium


(3) Standard solutions—Weigh accurately an amount ofSpectinomycin Hydrochloride Reference Standard, equiva-lent to about 20 mg (potency), dissolve in 0.1 mol/L phos-phate buŠer solution, pH 8.0 to make exactly 20 mL, anduse this solution as the standard stock solution. Keep thestandard stock solution at a temperature not exceeding 59Cand use within 10 days. Take exactly a suitable amount ofthe standard stock solution before use, add 0.1 mol/L phos-phate buŠer solution, pH 8.0 to make solutions so that eachmL contains 200 mg (potency) and 50 mg (potency), and usethese solutions as the high concentration standard solutionand the low concentration standard solution, respectively.

(4) Sample solutions—Weigh accurately an amount ofSpectinomycin Hydrochloride, equivalent to about 20 mg(potency), and dissolve in 0.1 mol/L phosphate buŠer solu-tion, pH 8.0 to make exactly 20 mL. Take exactly a suitableamount of this solution, add 0.1 mol/L phosphate buŠer so-lution, pH 8.0 to make solutions so that each mL contains200 mg (potency) and 50 mg (potency), and use these solu-tions as the high concentration sample solution and the lowconcentration sample solution, respectively.


Streptomycin Sulfate硫酸ストレプトマイシン


Streptomycin Sulfate contains not less than 740 mg(potency) per mg, calculated on the dried basis. Thepotency of Streptomycin Sulfate is expressed as mass(potency) of streptomycin (C21H39N7O12: 581.57).

Description Streptomycin Sulfate occurs as a white to lightyellowish white powder.

It is freely soluble in water, and very slightly soluble inethanol (95).

Identiˆcation (1) Dissolve 50 mg of Streptomycin Sulfatein 5 mL of water, add 1 mL of ninhydrin TS and 0.5 mL ofpyridine, and heat for 10 minutes: a purple color is devel-oped.

(2) Dissolve 10 mg each of Streptomycin Sulfate andStreptomycin Sulfate Reference Standard in 10 mL of water,and use these solutions as the sample solution and the stan-dard solution. Perform the test with these solutions as

directed under the Thin-layer Chromatography. Spot 10 mLeach of the sample solution and the standard solution on aplate of silica gel for thin-layer chromatography. Developthe plate with a solution of potassium dihydrogen phosphate(7 in 100) to a distance of about 12 cm, and air-dry the plate.Spray evenly a mixture of a solution of 1,3-dihydrox-ynaphthalene in ethanol (95) (1 in 500) and diluted sulfuricacid (1 in 5) (1:1) on the plate, and heat at about 1509C forabout 5 minutes: the principal spots from the sample solu-tion and the standard solution show the same in color toneand Rf value.

(3) A solution of Streptomycin Sulfate (1 in 5) respondsto the Qualitative Tests for sulfate.

Optical rotation [a]D20: －79 –－889(0.5 g calculated onthe dried basis, water, 50 mL, 100 mm).

pH The pH of a solution obtained by dissolving 2.0 g ofStreptomycin Sulfate in 10 mL of water is between 4.5 and7.0.

Purity (1) Clarity and color of solution—A solution ob-tained by dissolving 1.0 g of Streptomycin Sulfate in 5 mL ofwater is clear, and colorless or pale yellow.

(2) Heavy metals—Proceed with 2.0 g of StreptomycinSulfate according to Method 4, and perform the test. Pre-pare the control solution with 2.0 mL of Standard Lead So-lution (not more than 10 ppm).

(3) Arsenic—Prepare the test solution with 2.0 g ofStreptomycin Sulfate according to Method 3 and performthe test (not more than 1 ppm).

(4) Related substances—Dissolve exactly 0.20 g of Strep-tomycin Sulfate in a mixture of methanol and sulfuric acid(97:3) to make 5 mL, and heat under a re‰ux condenser for 1hour. After cooling, wash the inside of the condenser with asuitable amount of a mixture of methanol and sulfuric acid(97:3), add a mixture of methanol and sulfuric acid (97:3) tomake exactly 20 mL, and use this solution as the sample so-lution. Separately, dissolve exactly 36 mg of D(＋)-mannosein a mixture of methanol and sulfuric acid (97:3) to make 5mL, and heat under a re‰ux condenser for 1 hour. Aftercooling, wash the inside of the condenser with a suitableamount of a mixture of methanol and sulfuric acid (97:3),and add a mixture of methanol and sulfuric acid (97:3) tomake exactly 50 mL. Pipet 5 mL of this solution, add a mix-ture of methanol and sulfuric acid (97:3) to make exactly 50mL, and use this solution as the standard solution. Performthe test with these solutions as directed under the Thin-layerChromatography. Spot 10 mL each of the sample solutionand the standard solution on a plate of silica gel for thin-lay-er chromatography, develop with a mixture of toluene,methanol and acetic acid (100) (2:1:1) to a distance of 13 to15 cm, and air-dry the plate. Spray evenly a mixture of a so-lution of 1,3-dihydroxynaphthalene in ethanol (95) (1 in 500)and diluted sulfuric acid (1 in 5) (1:1) on the plate, and heatat 1109C for 5 minutes: the spot from the sample solutioncorresponding to the spot from the standard solution is notmore intense than the spot from the standard solution.

Loss on drying Not more than 5.0z (0.5 g, reduced pres-


sure not exceeding 0.67 kPa, 609C, 3 hours).




for test organism [5] under (1) Agar media for seed and baselayer, having pH 7.8 – 8.0 after sterilization.

(3) Standard solutions—Weigh accurately an amount ofStreptomycin Sulfate Reference Standard, previously dried,equivalent to about 20 mg (potency), dissolve in dilutedphosphate buŠer solution, pH 6.0 (1 in 2) to make exactly 50mL, and use this solution as the standard stock solution.Keep the standard stock solution between 59C and 159C,and use within 30 days. Take exactly a suitable amount ofthe standard stock solution before use, add 0.1 mol/L phos-phate buŠer solution, pH 8.0 to make solutions so that eachmL contains 8 mg (potency) and 2 mg (potency), and usethese solutions as the high concentration standard solutionand the low concentration standard solution, respectively.

(4) Sample solutions—Weigh accurately an amount ofStreptomycin Sulfate, equivalent to about 20 mg (potency),dissolve in water to make exactly 50 mL. Take exactly a suit-able amount of this solution, add 0.1 mol/L phosphatebuŠer solution, pH 8.0 to make solutions so that each mLcontains 8 mg (potency) and 2 mg (potency), and use these so-lutions as the high concentration sample solution and thelow concentration sample solution, respectively.


Sulbenicillin Sodiumスルベニシリンナトリウム


Sulbenicillin Sodium contains not less than 814 mg(potency) per mg, calculated on the anhydrous basis.The potency of Sulbenicillin Sodium is expressed asmass (potency) of sulbenicillin (C16H18N2O7S2:414.45).

Description Sulbenicillin Sodium occurs as white to lightyellowish white powder.

It is very soluble in water, freely soluble in methanol, andslightly soluble in ethanol (99.5).

It is hygroscopic.

Identiˆcation (1) Determine the infrared absorptionspectrum of Sulbenicillin Sodium as directed in the potassi-um bromide disk method under the Infrared Spectrophoto-metry, and compare the spectrum with the Reference Spec-trum or the spectrum of Sulbenicillin Sodium ReferenceStandard: both spectra exhibit similar intensities of absorp-

tion at the same wave numbers.(2) Sulbenicillin Sodium responds to the Qualitative Test

(1) for sodium salt.

Optical rotation [a]D20: ＋167 –＋1829(1 g calculated onthe anhydrous basis, water, 20 mL, 100 mm).

pH The pH of a solution obtained by dissolving 0.20 g ofSulbenicillin Sodium in 10 mL of water is between 4.5 and7.0.

Purity (1) Clarity and color of solution—Dissolve 2.5 gof Sulbenicillin Sodium in 5 mL of water: the solution isclear and colorless to pale yellow.

(2) Heavy metals—Proceed with 1.0 g of SulbenicillinSodium according to Method 1, and perform the test. Pre-pare the control solution with 2.0 mL of Standard Lead So-lution (not more than 20 ppm).

(3) Arsenic—Prepare the test solution with 1.0 g of Sul-benicillin Sodium according to Method 1, and perform thetest (not more than 2 ppm).

(4) Related substances—Dissolve 0.10 g of SulbenicillinSodium in 15 mL of the mobile phase, and use this solutionas the sample solution. Perform the test with 10 mL of thesample solution as directed under the Liquid Chro-matography according to the following conditions, deter-mine each peak area by the automatic integration method,and calculate the amount of these peaks by the area percen-tage method: the amount of the each peak other than thetwo peaks of sulbenicillin is not more than 2.0z, and thetotal amount of the peaks other than the two peaks of sul-benicillin is not more than 5.0z.Operating conditions—




Mobile phase: Dissolve 10 g of potassium dihydrogenphosphate in 750 mL of water, adjust the pH to 6.0± 0.1with sodium hydroxide TS, and add water to make 1000 mL.To 940 mL of this solution add 60 mL of acetonitrile.

Flow rate: Adjust the ‰ow rate so that the retention timeof the lately eluted peak of sulbenicillin is about 18 minutes.

Time span of measurement: About 1.5 times as long as theretention time of the lately eluted peak of sulbenicillin afterthe solvent peak.System suitability—

Test for required detectability: Measure exactly 1 mL ofthe sample solution, add the mobile phase to make exactly100 mL, and use this solution as the solution for systemsuitability test. Pipet 1 mL of the solution for systemsuitability test, and add the mobile phase to make exactly 10mL. Conˆrm that the total area of the two peaks of sul-benicillin obtained from 10 mL of this solution is equivalentto 7 to 13z of that from 10 mL of the solution for system


suitability test.System performance: When the procedure is run with 10

mL of the sample solution under the above operating condi-tions, the resolution between the two peaks of sulbenicillin isnot less than 2.0.

System repeatability: When the test is repeated 6 timeswith 10 mL of the solution for system suitability test underthe above operating conditions, the relative standard devia-tion of the total areas of the two peaks of sulbenicillin is notmore than 5.0z.




for test organism [5] under (1) Agar media for seed and baselayer. Adjust the pH of the medium so that it will be 6.4 to6.6 after sterilization.

(3) Standard solutions—Weigh accurately an amount ofSulbenicillin Sodium Reference Standard, equivalent toabout 50 mg (potency), dissolve in phosphate buŠer solu-tion, pH 6.0 to make exactly 50 mL, and use this solution asthe standard stock solution. Keep the standard stock solu-tion in a freezer, and use within 4 days. Take exactly a suita-ble amount of the standard stock solution before use, addphosphate buŠer solution, pH 6.0 to make solutions so thateach mL contains 40 mg (potency) and 10 mg (potency), anduse these solutions as the high concentration standard solu-tion and the low concentration standard solution, respec-tively.

(4) Sample solutions—Weigh accurately an amount ofSulbenicillin Sodium, equivalent to about 50 mg (potency),and dissolve in phosphate buŠer solution, pH 6.0 to makeexactly 50 mL. Take exactly a suitable amount of this solu-tion, add phosphate buŠer solution, pH 6.0 to make solu-tions so that each mL contains 40 mg (potency) and 10 mg(potency), and use these solutions as the high concentrationsample solution and the low concentration sample solution,respectively.


Talampicillin Hydrochloride塩酸タランピシリン


Talampicillin Hydrochloride is the hydrochloride ofampicillin phthalidyl ester.

It contains not less than 600 mg (potency) per mg,calculated on the anhydrous basis. The potency ofTalampicillin Hydrochloride is expressed as mass(potency) of ampicillin (C16H19N3O4S: 349.40).

Description Talampicillin Hydrochloride occurs as a whiteto light yellowish white powder.

It is very soluble in methanol, and freely soluble in waterand in ethanol (99.5).

Identiˆcation (1) To 1 mL of a solution of TalampicillinHydrochloride (1 in 30) add 1 mL of sodium hydroxide TS,mix, allow to stand for 5 minutes, and add 2 mL of dilutesulfuric acid and 2 to 3 drops of 2,4-dinitrophenylhydrazineTS: an orange-yellow precipitate is formed.

(2) Determine the infrared absorption spectrum ofTalampicillin Hydrochloride as directed in the potassiumbromide disk method under the Infrared Spectrophotomet-ry, and compare the spectrum with the Reference Spectrumor the spectrum of Talampicillin Hydrochloride ReferenceStandard: both spectra exhibit similar intensities of absorp-tion at the same wave numbers.

(3) To 10 mL of a solution of Talampicillin Hydrochlo-ride (1 in 300) add 1 mL of dilute nitric acid, and add silvernitrate TS: a white precipitate is formed.

Optical rotation [a]D20: ＋151 –＋1719(0.2 g calculated onthe anhydrous basis, ethanol (99.5), 20 mL, 100 mm).

Purity (1) Heavy metals—Proceed with 1.0 g of Talam-picillin Hydrochloride according to Method 2, and performthe test. Prepare the control solution with 2.0 mL of Stan-dard Lead Solution (not more than 20 ppm).

(2) Arsenic—Prepare the test solution with 1.0 g ofTalampicillin Hydrochloride according to Method 4, andperform the test (not more than 2 ppm).

(3) Related substances—Dissolve 50 mg of TalampicillinHydrochloride in ethanol (99.5) to make exactly 10 mL, anduse this solution as the sample solution. Pipet 1 mL, 2 mLand 3 mL of the sample solution, add ethanol (99.5) to eachto make exactly 100 mL, and use these solutions as the stan-dard solution (1), the standard solution (2) and the standardsolution (3), respectively. Perform the test with these solu-tions as directed under the Thin-layer Chromatography.Spot 10 mL each of the sample solution and the standard so-lutions (1), (2) and (3) on a plate of silica gel for thin-layerchromatography. Develop the plate with a mixture of tetra-hydrofuran, ethyl acetate, water and ethanol (95) (4:4:2:1)to a distance of about 13 cm, and air-dry the plate. Sprayevenly a solution of ninhydrin in ethanol (99.5) (1 in 500) on


the plate, and heat at 1109C for 5 minutes: the spots otherthan the principal spot from the sample solution is not moreintense than the spot from the standard solution (3), and thetotal of the amount of each spot other than the principalspot from the sample solution, which is calculated by thecomparison with the spots obtained from the standard solu-tions (1), (2) and (3), is not more than 5z.

(4) 2-Formylbenzoic acid—Dissolve 50 mg of Talam-picillin Hydrochloride in ethanol (99.5) to make exactly 10mL, and use this solution as the sample solution. Separately,dissolve 10 mg of 2-formylbenzoic acid in ethanol (99.5) tomake exactly 100 mL. Pipet 5 mL of this solution, addethanol (99.5) to make exactly 10 mL, and use this solutionas the standard solution. Perform the test with these solu-tions as directed under the Thin-layer chromatography. Spot10 mL each of the sample solution and the standard solutionon a plate of silica gel for thin-layer chromatography, de-velop the plate with a mixture of chloroform and acetic acid(100) (4:1) to a distance of about 13 cm, and air-dry theplate. Spray evenly a solution of 2,4-dinitrophenylhydrazinein diluted sulfuric acid (6 in 25) (1 in 500): the spot of 2-for-mylbenzoic acid obtained from the sample solution is notmore intense than that obtained from the standard solution.


Assay Weigh accurately an amount of TalampicillinHydrochloride and Talampicillin Hydrochloride ReferenceStandard, equivalent to about 20 mg (potency), dissolve inwater to make exactly 20 mL each, and use these solutions asthe sample solution and the standard solution. The standardsolution should be prepared before use. Pipet 2 mL each ofthe sample solution and the standard solution in separate100-mL glass-stoppered ‰asks, add 2.0 mL of sodiumhydroxide TS, and allow them to stand for exactly 15minutes. Add 2.0 mL of diluted hydrochloric acid (1 in 10)and exactly 10 mL of 0.005 mol/L iodine VS, allow them tostand for exactly 15 minutes, and titrate with 0.01 mol/L so-dium thiosulfate VS until the color of the solution is disap-peared. If necessary, add 0.2 to 0.5 mL of starch TS.Separately, pipet 2 mL each of the sample solution and thestandard solution in separate 100-mL glass-stoppered ‰asks,add exactly 10 mL of 0.005 mol/L iodine VS, titrate with0.01 mol/L sodium thiosulfate VS until the color of the so-lution is disappeared, and make any necessary correction.For this titration, add 0.2 to 0.5 mL of starch TS, if necessa-ry. Calculate the amount (mL) of 0.005 mol/L iodine VS, VT

and VS, consumed by the sample solution and the standardsolution, respectively.


＝WS×

VT

VS× 1000

WS: Amount [mg (potency)] of Talampicillin Hydrochlo-ride Reference Standard


Tegafurテガフール


Description Tegafur occurs as a white, crystalline powder.It is soluble in methanol and in acetone, and sparingly

soluble in water and in ethanol (95).It dissolves in dilute sodium hydroxide TS.A solution of Tegafur in methanol (1 in 50) shows no opti-

cal rotation.


Purity(4) Arsenic—Prepare the test solution in a platinum

crucible with 1.0 g of Tegafur according to Method 4, in-cinerating by ignition between 7509C and 8509C, and per-form the test (not more than 2 ppm).

Teicoplaninテイコプラニン


Teicoplanin contains not less than 900 mg (potency)per mg, calculated on the anhydrous, de-sodium chlo-ride and de-residual solvents basis. The potency ofTeicoplanin is expressed as mass (potency) of teicopla-nin (C72-89H68-99Cl2N8-9O28-33).

Change the Content ratio of the active principleto read:

Content ratio of the active principle Dissolve about 20 mgof Teicoplanin in water to make 10 mL, and use this solutionas the sample solution. Perform the test with 20 mL of thesample solution as directed under the Liquid Chro-matography according to the following conditions, and cal-culate the sum of peak areas of teicoplanin A2 group, Sa, thesum of peak areas of teicoplanin A3 group, Sb, and the sumof peak areas of other contents, Sc from the sample solutionby the automatic integration method. Calculate the contentratio of them by the formula given below: teicoplanin A2

group, teicoplanin A3 group, and the other are not less than80.0z, not more than 15.0z and not more than 5.0z, re-spectively.

The elution order of each content and the relative reten-tion time of each content to the retention time of teicoplaninA2-2 are shown in the following table.


Name of content Elutionorder

Relativeretention time

teicoplanin A3 group ≦0.42teicoplanin A3-1 1 0.29

teicoplanin A2 group 0.42＜,≦1.25teicoplanin A2-1 2 0.91teicoplanin A2-2 3 1.00teicoplanin A2-3 4 1.04teicoplanin A2-4 5 1.17teicoplanin A2-5 6 1.20

others 1.25＜

Content ratio (z) of teicoplanin A2 group

＝Sa

Sa＋ 0.83Sb＋ Sc× 100

Content ratio (z) of teicoplanin A3 group

＝0.83Sb

Sa＋ 0.83Sb＋ Sc× 100

Content ratio (z) of others

＝Sc

Sa＋ 0.83Sb＋ Sc× 100





Mobile phase A: Dissolve 7.80 g of sodium dihydrogenphosphate dihydrate in 1650 mL of water, add 300 mL ofacetonitrile, adjust pH to 6.0 with sodium hydroxide TS,and add water to make 2000 mL.

Mobile phase B: Dissolve 7.80 g of sodium dihydrogenphosphate dihydrate in 550 mL of water, add 1400 mL ofacetonitrile, adjust the pH to 6.0 with sodium hydroxide TS,and add water to make 2000 mL.

Flowing of the mobile phase: Flow mobile phase A for 10minutes before injection. After injection, control thegradient by mixing the mobile A and B as directed in the fol-lowing table.

Time after injection Mobile phase Mobile phaseof sample (min) A (z) B (z)

0 – 32 100→ 70 0→ 3032 – 40 70→ 50 30→ 5040 – 42 50→ 100 50→ 0

Flow rate: 1.8 mL per minute.Time span of measurement: About 1.7 times as long as the

retention time of teicoplanin A2-2 after the solvent peak.System suitability—

Test for required detection: Conˆrm that peak height ofteicoplanin A2-2 obtained from the sample solution is equiva-lent to 90z of the full scale.

System performance: When the procedure is run with 20mL of the sample solution under the above operating condi-tions, the symmetry coe‹cient of the peak of teicoplaninA3-1 is not more than 2.2.

Change the Bacterial endotoxins to read:

Bacterial endotoxins Less than 0.75 EU/mg (potency).

Theophyllineテオフィリン


Description Theophylline occurs as white crystals or crys-talline powder.

It is soluble in N,N-dimethylformamide, and slightly solu-ble in water and in ethanol (99.5).

It dissolves in 0.1 mol/L hydrochloric acid TS.


Identiˆcation (1) Determine the absorption spectrum ofa solution of Theophylline in 0.1 mol/L hydrochloric acidTS (1 in 200,000) as directed under the Ultraviolet-visibleSpectrophotometry, and compare the spectrum with theReference Spectrum: both spectra exhibit similar intensitiesof absorption at the same wavelengths.

(2) Determine the infrared absorption spectrum of The-ophylline, previously dried, as directed in the potassiumbromide disk method under the Infrared Spectrophotomet-ry, and compare the spectrum with the Reference Spectrum:both spectra exhibit similar intensities of absorption at thesame wave numbers.

Delete the Purity (5).


Assay Weigh accurately about 0.25 g of Theophylline,previously dried, and dissolve in 100 mL of water, add ex-actly 20 mL of 0.1 mol/L silver nitrate VS, shake the mix-ture, and titrate with 0.1 mol/L sodium hydroxide VS(potentiometric titration). Perform a blank determination,and make any necessary correction.

Each mL of 0.1 mol/L sodium hydroxide VS＝ 18.02 mg of C7H8N4O2


Thiamine Hydrochloride塩酸チアミン


Water Not more than 5.0z (30 mg, coulometric titration).

Thiamine Nitrate硝酸チアミン


Assay Weigh accurately about 0.1 g each of Thiamine Ni-trate, previously dried, and Thiamine Hydrochloride Refer-ence Standard (previously determine its water content in thesame manner as directed under Thiamine Hydrochloride),and dissolve them in the mobile phase to make exactly 50mL. To 10 mL each of the solutions, accurately measured,add exactly 5 mL each of the internal standard solution, addthe mobile phase to make 50 mL, and use these solutions asthe sample solution and the standard solution. Perform thetest with 10 mL each of the sample solution and the standardsolution as directed under the Liquid Chromatography ac-cording to the following conditions and calculate the ratios,QT and QS, of the peak area of thiamine to that of the inter-nal standard.

Amount (mg) of C12H17N5O4S＝WS×QT

QS× 0.9706

WS: Amount (mg) of Thiamine Hydrochloride ReferenceStandard, calculated on the anhydrous basis

Internal standard solution—A solution of methyl benzoatein methanol (1 in 50).Operating conditions—

Detector: An ultraviolet spectrophotometer (wavelength:254 nm).



Mobile phase: Dissolve 1.1 g of sodium l-octanesulfonatein 1000 mL of diluted acetic acid (100) (1 in 100). To 600 mLof this solution add 400 mL of a mixture of methanol andacetonitrile (3:2).

Flow rate: Adjust the ‰ow rate so that the retention timeof thiamine is about 12 minutes.System suitability—

System performance: When the procedure is run with 10mL of the standard solution under the above operating con-ditions, thiamine and the internal standard are eluted in thisorder with the resolution between these peaks being not lessthan 6.

System repeatability: When the test is repeated 6 times

with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratiosof the peak area of thiamine to that of the internal standardis not more than 1.0z.

Thiopental Sodiumチオペンタールナトリウム


Purity(4) Related substances—Dissolve 50 mg of Thiopental

Sodium in 50 mL of the mobile phase, and use this solutionas the sample solution. Pipet 1 mL of the sample solution,add the mobile phase to make exactly 200 mL, and use thissolution as the standard solution. Perform the test with ex-actly 20 mL each of the sample solution and the standard so-lution as directed under the Liquid Chromatography accord-ing to the following conditions. Measure each peak area ofeach solution by the automatic integration method: the totalarea of peaks other than those of thiopental in the samplesolution is not larger than the peak area of thiopental in thestandard solution.Operating conditions—


Column: A stainless steel column 4.6 mm in inside di-ameter and 15 cm in length, packed with octadecylsilanizedsilica gel (5 mm in particle diameter).


Mobile phase: Dissolve 1 g of potassium dihydrogen phos-phate in 1000 mL of water, and adjust the pH to 3.0 withphosphoric acid. To 700 mL of this solution add 300 mL ofacetonitrile.

Flow rate: Adjust the ‰ow rate so that the retention timeof thiopental is about 15 minutes.

Time span of measurement: About 1.5 times as long as theretention time of thiopental.System suitability—

Test for required detection: To exactly 2 mL of the stan-dard solution add the mobile phase to make exactly 10 mL.Conˆrm that the peak area of thiopental obtained from 20mL of this solution is equivalent to 15 to 25z of that ofthiopental obtained from 20 mL of the standard solution.

System performance: Dissolve 5 mg each of isopropylparahydroxybenzoate and propyl parahydroxybenzoate in50 mL of acetonitrile, and add water to make 100 mL. Whenthe procedure is run with 20 mL of this solution under theabove operating conditions, isopropyl parahydroxybenzoateand propyl parahydroxybenzoate are eluted in this orderwith the resolution between these peaks being not less than1.9.

System repeatability: When the test is repeated 6 timeswith 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak


area of thiopental is not more than 2.0z.

Tipepidine Hibenzateヒベンズ酸チペピジン


Purity(4) Related substances—(i) Dissolve 10 mg of Tipep-

idine Hibenzate in 20 mL of the mobile phase, and use thissolution as the sample solution. Pipet 1 mL of the sample so-lution, add the mobile phase to make exactly 100 mL, anduse this solution as the standard solution. Perform the testwith exactly 20 mL each of the sample solution and the stan-dard solution as directed under the Liquid Chromatographyaccording to the following conditions. Determine each peakarea of both solutions by the automatic integration method:the total area of all peaks other than the area of the hibenzicacid and tipepidine from the sample solution is not largerthan the peak area of the tipepidine from the standard solu-tion.Operating conditions—




Mobile phase: A mixture of a solution of ammoniumacetate (1 in 100) and tetrahydrofuran (32:13).

Flow rate: Adjust the ‰ow rate so that the retention timeof tipepidine is about 12 minutes.

Time span of measurement: As long as the retention timeof tipepidine after the solvent peak.System suitability—

Test for required detection: To exactly 2 mL of the stan-dard solution add the mobile phase to make exactly 20 mL.Conˆrm that the peak area of tipepidine obtained from 20mL of this solution is equivalent to 7 to 13z of that of tipepi-dine obtained from 20 mL of the standard solution.

System performance: Dissolve 10 mg of TipepidineHibenzate and 3 mg of propyl parahydroxybenzoate in 100mL of the mobile phase. When the procedure is run with 20mL of this solution under the above operating conditions,hibenzic acid, tipepidine and propyl parahydroxybenzoateare eluted in this order with the resolution between the peaksof tipepidine and propyl parahydroxybenzoate being not lessthan 3.

System repeatability: When the test is repeated 6 timeswith 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peakarea of tipepidine is not more than 1.5z.

(ii) Dissolve 10 mg of Tipepidine Hibenzate in 20 mL ofthe mobile phase, and use this solution as the sample solu-

tion. Pipet 1 mL of the sample solution, add the mobilephase to make exactly 100 mL, and use this solution as thestandard solution. Perform the test with exactly 20 mL eachof the sample solution and the standard solution as directedunder the Liquid Chromatography according to the follow-ing conditions, and determine each peak area by the auto-matic integration method: the total area of all peaks otherthan the area of the hibenzic acid and tipepidine from thesample solution is not larger than 1W2 times the peak area ofthe tipepidine from the standard solution.Operating conditions—




Mobile phase: A mixture of methanol and a solution ofammonium acetate (1 in 500) (13:7).

Flow rate: Adjust the ‰ow rate so that the retention timeof tipepidine is about 10 minutes.

Time span of measurement: As long as the retention timeof tipepidine after the solvent peak.System suitability—

Test for required detection: To exactly 2 mL of the stan-dard solution add the mobile phase to make exactly 20 mL.Conˆrm that the peak area of tipepidine obtained from 20mL of this solution is equivalent to 7 to 13z of that of tipepi-dine obtained from 20 mL of the standard solution.

System performance: Dissolve 12 mg of TipepidineHibenzate and 4 mg of xanthene in 50 mL of the mobilephase. When the procedure is run with 10 mL of this solutionunder the above operating conditions, hibenzic acid, tipepi-dine and xanthene are eluted in this order with the resolutionbetween the peaks of tipepidine and xanthene being not lessthan 3.

System repeatability: When the test is repeated 6 timeswith 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peakarea of tipepidine is not more than 3.0z.

Tobramycinトブラマイシン


Tobramycin contains not less than 900 mg (potency)per mg, calculated on the anhydrous basis. The poten-cy of Tobramycin is expressed as mass (potency) oftobramycin (C18H37N5O9).

Description Tobramycin occurs as a white to pale yellow-ish white powder.


It is very soluble in water, freely soluble in formamide,slightly soluble in methanol, and very slightly soluble inethanol (95).

It is hygroscopic.

Identiˆcation (1) Determine the spectrum of a solutionof Tobramycin in heavy water for nuclear magneticresonance spectroscopy (1 in 125) as directed under theNuclear Magnetic Resonance Spectroscopy (1H), using sodi-um 3-trimethylsilylpropanesulfonate for nuclear magneticresonance spectroscopy as an internal reference compound:it exhibits a double signal A at around d5.1 ppm, a multiplesignal B between d2.6 ppm and d4.0 ppm, and a multiplesignal C between d1.0 ppm and d2.1 ppm. The ratio of theintegrated intensity of these signals, A:B:C, is about 1:8:2.

(2) Dissolve 10 mg each of Tobramycin and TobramycinReference Standard in 1 mL of water, and use these solu-tions as the sample solution and the standard solution.Perform the test with these solutions as directed under theThin-layer Chromatography. Spot 4 mL of the sample solu-tion and the standard solution on a plate of silica gel forthin-layer chromatography. Develop the plate with a mix-ture of ammonia TS, 1-butanol and methanol (5:5:2) to adistance of about 10 cm, and air-dry the plate. Spray evenlyninhydrin TS on the plate, and heat at 1009C for 5 minutes:the Rf values of the principal spots obtained from the sam-ple solution and the standard solution are the same.



pH The pH of a solution obtained by dissolving 0.10 g ofTobramycin in 10 mL of water is between 9.5 and 11.5.

Purity (1) Clarity and color of solution—Dissolve 1.0 gof Tobramycin in 10 mL of water: the solution is clear andcolorless to pale yellow.

(2) Heavy metals—Proceed with 1.0 g of Tobramycinaccording to Method 2, and perform the test. Prepare thecontrol solution with 3.0 mL of Standard Lead Solution (notmore than 30 ppm).

(3) Related substances—Dissolve 80 mg of Tobramycinin 10 mL of diluted ammonia solution (28) (1 in 250), anduse this solution as the sample solution. Pipet 1 mL of thesample solution, add diluted ammonia solution (28) (1 in250) to make exactly 100 mL, and use this solution as thestandard solution. Perform the test with these solutions asdirected under the Thin-layer Chromatography. Spot 5 mLof the sample solution and the standard solution on a plateof silica gel for thin-layer chromatography. Develop theplate with a mixture of ammonia solution (28), ethanol (95)and 2-butanone (1:1:1) to a distance of about 10 cm, air-drythe plate, then further dry at 1109C for 10 minutes. Immedi-ately spray evenly a mixture of water and sodiumhypochlorite TS (4:1) on the plate, air-dry the plate, thenspray potassium iodide-starch TS on the plate: the spot otherthan the principal spot from the sample solution is not moreintense than the spot from the standard solution.

Water Not more than 11.0z (0.1 g, volumetric titration,

direct titration). Use a mixture of formamide for water de-termination and methanol for water determination (3:1) in-stead of methanol for water determination.





(3) Standard solutions—Weigh accurately an amount ofTobramycin Reference Standard, equivalent to about 25 mg(potency), dissolve in 0.1 mol/L phosphate buŠer solution,pH 8.0 to make exactly 25 mL, and use this solution as thestandard stock solution. Keep the standard stock solutionbetween 59C and 159C, and use within 30 days. Take exactlya suitable amount of the standard stock solution before use,add 0.1 mol/L phosphate buŠer solution, pH 8.0 to makesolutions so that each mL contains 8 mg (potency) and 2 mg(potency), and use these solutions as the high concentrationstandard solution and the low concentration standard solu-tion, respectively.

(4) Sample solutions—Weigh accurately an amount ofTobramycin, equivalent to about 25 mg (potency), and dis-solve in 0.1 mol/L phosphate buŠer solution, pH 8.0 tomake exactly 25 mL. Take exactly a suitable amount of thissolution, add 0.1 mol/L phosphate buŠer solution, pH 8.0to make solutions so that each mL contains 8 mg (potency)and 2 mg (potency), and use these solutions as the high con-centration sample solution and the low concentration samplesolution, respectively.


Tocopherolトコフェロール


Assay Dissolve about 50 mg each of Tocopherol and To-copherol Reference Standard, accurately weighed, in etha-nol (99.5) to make exactly 50 mL, and use these solutions asthe sample solution and the standard solution. Perform thetest with exactly 20 mL each of these solutions as directed un-der the Liquid Chromatography according to the followingconditions, and determine the peak heights, HT and HS, oftocopherol in the sample solution and the standard solution.

Amount (mg) of C29H50O2＝WS×HT

HS

WS: Amount (mg) of Tocopherol Reference Standard


length: 292 nm).


Column: A stainless steel column 4.6 mm in inside di-ameter and 15 cm in length, packed with octadecylsilanizedsilica gel for liquid chromatography (5 mL in particlediameter).



of tocopherol is about 10 minutes.System suitability—

System performance: Dissolve 0.05 g each of Tocopheroland tocopherol acetate in 50 mL of ethanol (99.5). When theprocedure is run with 20 mL of this solution under the aboveoperating conditions, tocopherol and tocopherol acetate areeluted in this order with the resolution between these peaksbeing not less than 2.6.

System repeatability: When the test is repeated 5 timeswith 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peakheights of tocopherol is not more than 0.8z.

Tocopherol Acetate酢酸トコフェロール


Assay Dissolve 50 mg each of Tocopherol Acetate andTocopherol Acetate Reference Standard, accuratelyweighed, in ethanol (99.5) to make exactly 50 mL, and usethese solutions as the sample solution and the standard solu-tion. Perform the test with exactly 20 mL each of these solu-tions as directed under the Liquid Chromatography accord-ing to the following conditions, and determine the peakheights, HT and HS, of tocopherol acetate in the sample solu-tion and the standard solution, respectively.

Amount (mg) of C31H52O3＝WS×HT

HS

WS: Amount (mg) of Tocopherol Acetate ReferenceStandard



ameter and 15 cm in length, packed with octadecylsilanizedsilica gel (5 mm in particle diameter).



of tocopherol acetate is about 12 minutes.System suitability—

System performance: Dissolve 0.05 g each of TocopherolAcetate and tocopherol in 50 mL of ethanol (99.5). Whenthe procedure is run with 20 mL of this solution under theabove operating conditions, tocopherol and tocopherol

acetate are eluted in this order with the resolution betweenthese peaks being not less than 2.6.

System repeatability: When the test is repeated 5 timeswith 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peakheights of tocopherol acetate is not more than 0.8z.

Triamcinoloneトリアムシノロン


Assay Dissolve about 20 mg each of Triamcinolone andTriamcinolone Reference Standard, previously dried and ac-curately weighed, in a solution of L-ascorbic acid in metha-nol (1 in 1000) to make exactly 50 mL. Pipet 5 mL each ofthese solutions, add exactly 5 mL each of the internal stan-dard solution, add a solution of L-ascorbic acid in methanol(1 in 1000) to make 20 mL, and use these solutions as thesample solution and the standard solution. Perform the testwith 10 mL each of these solutions as directed under the Liq-uid Chromatography according to the following conditions,and calculate the ratios, QT and QS, of the peak height of tri-amcinolone to that of the internal standard, respectively.

Amount (mg) of C21H27FO6＝WS×QT

QS

WS: Amount (mg) of Triamcinolone Reference Standard

Internal standard solution—Dissolve 15 mg of methyl para-hydroxybenzoate in a solution of L-ascorbic acid in metha-nol (1 in 1000) to make 100 mL.Operating conditions—


Column: A stainless steel column 4.0 mm in inside di-ameter and 15 cm in length, packed with octadecylsilanizedsilica gel (5 mm in particle diameter).



of triamcinolone is about 10 minutes.System suitability—

System performance: When the procedure is run with 10mL of the standard solution under the above operating con-ditions, triamcinolone and the internal standard are eluted inthis order with the resolution between these peaks being notless than 2.0.

System repeatability: When the test is repeated 6 timeswith 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratiosof the peak height of triamcinolone to that of the internalstandard is not more than 1.5z.


Change to read:

Trichomycinトリコマイシン

Trichomycin A33-(3-Amino-3,6-dideoxy-b-D-mannopyranosyloxy)-17-[6-(4-aminophenyl)-4-hydroxy-1-methyl-6-oxohexyl]-1,3,5,9,11,37-hexahydroxy-18-methyl-13,15-dioxo-16,39-dioxabicyclo[33.3.1]nonatriaconta-19,21,23,25,27,29,31-heptaene-36-carboxylic acid [12698-99-6]Trichomycin B33-(3-Amino-3,6-dideoxy-b-D-mannopyranosyloxy)-17-[6-(4-aminophenyl)-4-hydroxy-1-methyl-6-oxohexyl]-1,3,5,7,9,37-hexahydroxy-18-methyl-13,15-dioxo-16,39-dioxabicyclo[33.3.1]nonatriaconta-19,21,23,25,27,29,31-heptaene-36-carboxylic acid [12699-00-2][1394-02-1, Trichomycin]

Trichomycin contains not less than 7000 Units permg, calculated on the dried basis. The potency ofTrichomycin is expressed as unit based on the amountof trichomycin. 1 unit of Trichomycin is equivalent to0.05 mg of trichomycin.

Description Trichomycin occurs as a yellow to yellow-brown powder.

It is practically insoluble in water, in ethanol (99.5) and intetrahydrofuran.

It dissolves in dilute sodium hydroxide TS.It is hygroscopic.

Identiˆcation (1) To 2 mg of Trichomycin add 2 mL ofsulfuric acid: a blue color appears, and the color is changedto a blue-purple after allowing to stand.

(2) Dissolve 1 mg of Trichomycin in 50 mL of a solutionof sodium hydroxide (1 in 200). Determine the absorptionspectrum of this solution as directed under the Ultraviolet-visible Spectrophotometry: it exhibits maxima between 359nm and 365 nm, between 378 nm and 384 nm, and between400 nm and 406 nm.

Content ratio of the active principle Conduct this proce-dure without exposure to daylight, using light-resistant ves-sels. Dissolve about 10 mg of Trichomycin in 50 mL of amixture of tetrahydrofuran for liquid chromatography andwater (3:1), and use this solution as the sample solution. Per-form the test with 5 mL of the sample solution as directed un-der the Liquid Chromatography according to the followingconditions, determine the peak areas by the automatic in-tegration method, and calculate the amount of trichomycinA and trichomycin B by the area percentage method: the

amount of trichomycin A is between 20z and 40z, and thatof trichomycin B is between 15z and 25z. The relativeretention time of trichomycin B with respect to trichomycinA is about 1.2.Operating conditions—




Mobile phase: Dissolve 3.4 g of potassium dihydrogenphosphate and 1.7 g of sodium lauryl sulfate in a mixture of600 mL of water and 400 mL of acetonitrile for liquid chro-matography.

Flow rate: Adjust the ‰ow rate so that the retention timeof trichomycin A is about 8 minutes.

Time span of measurement: About 4 times as long as theretention time of trichomycin A.System suitability—

Test for required detectability: Measure exactly 5 mL ofthe sample solution, add a mixture of tetrahydrofuran forliquid chromatography and water (3:1) to make exactly 50mL, and use this solution as the solution for system suitabil-ity test. Pipet 5 mL of the solution for system suitability test,and add a mixture of tetrahydrofuran for liquid chro-matography and water (3:1) to make exactly 30 mL. Con-ˆrm that the peak area of trichomycin A obtained from 5 mLof this solution is equivalent to 12 to 22z of that from 5 mLof the solution for system suitability test.

System performance: When the procedure is run with 5 mLof the solution for system suitability test under the aboveoperating conditions, trichomycin A and trichomycin B areeluted in this order with the resolution between these peaksbeing not less than 2.5.

System repeatability: When the test is repeated 6 timeswith 5 mL of the solution for system suitability test under theabove operating conditions, the relative standard deviationof the peak area of trichomycin A is not more than 2.0z.


Assay Conduct this procedure without exposure to day-light, using light-resistant vessels. Weigh accurately anamount of Trichomycin and Trichomycin Reference Stan-dard, equivalent to about 150,000 units, dissolve themseparately in a mixture of tetrahydrofuran for liquid chro-matography and water (3:1) to make exactly 100 mL, anduse these solutions as the sample solution and the standardsolution. Perform the test with exactly 20 mL each of thesample solution and the standard solution as directed underthe Liquid Chromatography according to the following con-ditions, and determine the peak areas, AT and AS, oftrichomycin.


Amount (unit) of trichomycin＝WS×AT

AS

WS: Amount (unit) of Trichomycin Reference Standard



ameter and 15 cm in length, packed with silica gel for liquidchromatography (10 mm in particle diameter).


Mobile phase: Dissolve 15 g of ammonium acetate in 120mL of water, and add 1000 mL of acetonitrile for liquidchromatography and 700 mL of methanol.

Flow rate: Adjust the ‰ow rate so that the retention timeof trichomycin is about 6 minutes.System suitability—

System performance: Dissolve 5 mg of Trichomycin and 1mg of berberine chloride in 100 mL of a mixture of tetra-hydrofuran for liquid chromatography and water (3:1).When the procedure is run with 20 mL of this solution underthe above operating conditions, berberine and trichomycinare eluted in this order with the resolution between thesepeaks being not less than 4.

System repeatability: When the test is repeated 6 timeswith 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peakarea of trichomycin is not more than 2.0z.

Containers and storage Containers—Tight containers.Storage—Light-resistant, and in a cold place.

Add the following:

Trimebutine Maleateマレイン酸トリメブチン

C22H29NO5.C4H4O4: 503.54(2RS)-2-Dimethylamino-2-phenylbutyl 3,4,5-trimethoxybenzoate monomaleate [34140-59-5]

Trimebutine Maleate, when dried, contains not lessthan 98.5z and not more than 101.0z of trimebutinemaleate (C22H29NO5.C4H4O4).

Description Trimebutine Maleate occurs as white, crystalsor crystalline powder.

It is freely soluble in N,N-dimethylformamide and in acet-ic acid (100), soluble in acetonitrile, and slightly soluble inwater and in ethanol (99.5).

It dissolves in 0.01 mol/L hydrochloric acid TS.

A solution of it in N,N-dimethylformamide (1 in 20)shows no optical rotation.

Identiˆcation (1) Determine the absorption spectrum ofa solution of Trimebutine Maleate in 0.01 mol/Lhydrochloric acid TS (1 in 50,000) as directed under theUltraviolet-visible Spectrophotometry, and compare thespectrum with the Reference Spectrum: both spectra exhibitsimilar intensities of absorption at the same wavelengths.

(2) Determine the infrared absorption spectrum ofTrimebutine Maleate as directed in the potassium bromidedisk method under the Infrared Spectrophotometry, andcompare the spectrum with the Reference Spectrum: bothspectra exhibit similar intensities of absorption at the samewave numbers.


Purity (1) Heavy metals—Proceed with 2.0 g of Trime-butine Maleate according to Method 2, and perform the test.Prepare the control solution with 2.0 mL of Standard LeadSolution (not more than 10 ppm).

(2) Arsenic—Prepare the test solution with 2.0 g ofTrimebutine Maleate according to Method 3, and performthe test (not more than 1 ppm).

(3) Related substances—Dissolve 0.10 g of TrimebutineMaleate in 100 mL of a mixture of 0.01 mol/L hydrochloricacid TS and acetonitrile (13:7), and use this solution as thesample solution. Pipet 1 mL of the sample solution, add amixture of 0.01 mol/L hydrochloric acid TS and acetonitrile(13:7) to make exactly 250 mL, and use this solution as thestandard solution. Perform the test with exactly 20 mL eachof the sample solution and the standard solution as directedunder the Liquid Chromatography according to the follow-ing conditions, and determine each peak area by the auto-matic integration method: the area of the peak other thanmaleic acid and trimebutine from the sample solution is notmore than 1/2 times the peak area of trimebutine from thestandard solution, and the total area of the peaks other thanmaleic acid and trimebutine is not more than the peak areaof trimebutine from the standard solution.Operating conditions—




Mobile phase: To 650 mL of diluted perchloric acid (17 in20,000), previously adjusted the pH to 3.0 with a solution ofammonium acetate (1 in 1000), add 1 g of sodium 1-pen-tanesulfonate to dissolve. To 650 mL of this solution add350 mL of acetonitrile.

Flow rate: Adjust the ‰ow rate so that the retention timeof trimebutine is about 9 minutes.

Time span of measurement: About 2 times as long as theretention time of trimebutine after the peak of maleic acid.



the standard solution, and add a mixture of 0.01 mol/Lhydrochloric acid TS and acetonitrile (13:7) to make exactly20 mL. Conˆrm that the peak area of trimebutine obtainedfrom 20 mL of this solution is equivalent to 20 to 30z of thatfrom 20 mL of the standard solution.

System performance: Dissolve 40 mg of TrimebutineMaleate and 20 mg of imipramine hydrochloride in 100 mLof a mixture of 0.01 mol/L hydrochloric acid TS andacetonitrile (13:7). When the procedure is run with 20mL ofthis solution under the above operating conditions, trimebu-tine and imipramine are eluted in this order with the resolu-tion between these peaks being not less than 2.5.

System repeatability: When the test is repeated 6 timeswith 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peakarea of trimebutine is not more than 5z.



Assay Weigh accurately about 0.8 g of Trimebutine Male-ate, previously dried, dissolve in 70 mL of acetic acid (100),and titrate with 0.1 mol/L perchloric acid VS until the colorof the solution changes from purple through blue to blue-green (indicator: 3 drops of crystal violet TS). Perform ablank determination, and make any necessary correction.

Each mL of 0.1 mol/L perchloric acid VS＝ 50.35 mg of C22H29NO5.C4H4O4


Trimetoquinol Hydrochloride塩酸トリメトキノール


Trimetoquinol Hydrochloride contains not less than98.5z and not more than 101.0z of C19H23NO5.HCl(mol. wt.: 381.85), calculated on the anhydrous basis.


Description Trimetoquinol Hydrochloride occurs as white,crystals or crystalline powder.

It is freely soluble in methanol, and sparingly soluble inwater and in ethanol (99.5).

Melting point: about 1519C (with decomposition, afterdrying in vacuum, 1059C, 4 hours).


Identiˆcation (1) Determine the absorption spectrum ofa solution of Trimetoquinol Hydrochloride in 0.01 mol/Lhydrochloric acid TS (1 in 20,000) as directed under the

Ultraviolet-visible Spectrophotometry, and compare thespectrum with the Reference Spectrum: both spectra exhibitsimilar intensities of absorption at the same wavelengths.

(2) Determine the infrared absorption spectrum ofTrimetoquinol Hydrochloride as directed in the potassiumchloride disk method under the Infrared Spectrophotomet-ry, and compare the spectrum with the Reference Spectrum:both spectra exhibit similar intensities of absorption at thesame wave numbers.

(3) A solution of Trimetoquinol Hydrochloride (1 in 50)responds to the Qualitative Tests (1) for chloride.


Optical rotation [a]D20: －16 –－199(0.25 g, calculated onthe anhydrous basis, water, after warming and cooling,25 mL, 100 mm).


Purity (1) Clarity and color of solution—Dissolve 0.10 gof Trimetoquinol Hydrochloride in 10 mL of water bywarming: the solution is clear and colorless.

(2) Sulfate—Perform the test with 0.5 g of Trimeto-quinol Hydrochloride. Prepare the control solution with0.40 mL of 0.005 mol/L sulfuric acid VS (not more than0.038z).

(3) Heavy metals—Proceed with 1.0 g of TrimetoquinolHydrochloride according to Method 2, and perform the test.Prepare the control solution with 2.0 mL of Standard LeadSolution (not more than 20 ppm).

(4) Related substances—Dissolve 50 mg of Trimeto-quinol Hydrochloride in 50 mL of the mobile phase, and usethis solution as the sample solution. Pipet 1 mL of this solu-tion, add the mobile phase to make exactly 100 mL, and usethis solution as the standard solution. Perform the test withexactly 20 mL each of the sample solution and the standardsolution, as directed under the Liquid Chromatography ac-cording to the following conditions. Determine each peakarea of both solutions by the automatic integration method:the total area of the peaks other than that of trimetoquinolfrom the sample solution is not larger than the peak area oftrimetoquinol from the standard solution.Operating conditions—




Mobile phase: Dissolve 2 g of potassium dihydrogen phos-phate and 2 g of sodium 1-pentane sulfonate in 1000 mL ofwater. Adjust with phosphoric acid to a pH between 2.8 and3.2, and ˆlter through a membrane ˆlter with pore size of 0.4mm. Add 200 mL of acetonitrile to 800 mL of the ˆltrate.

Flow rate: Adjust the ‰ow rate so that the retention timeof trimetoquinol is about 7 minutes.


Time span of measurement: About twice as long as theretention time of trimetoquinol after the solvent peak.System suitability—

Test for required detection: To exactly 2 mL of the stan-dard solution add the mobile phase to make exactly 20 mL.Conˆrm that the peak area of trimetoquinol obtained from20 mL of this solution is equivalent to 7 to 13z of that oftrimetoquinol obtained from 20 mL of the standard solution.

System performance: Dissolve 5 mg of TrimetoquinolHydrochloride and 1 mg of procaine hydrochloride in 50 mLof the mobile phase. When the procedure is run with 20 mLof this solution under the above operating conditions,procaine and trimetoquinol are eluted in this order with theresolution between these peaks being not less than 4.

System repeatability: When the test is repeated 6 timeswith 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peakarea of trimetoquinol is not more than 2.0z.

Delete the Loss on drying and add the following:


Vancomycin Hydrochloride塩酸バンコマイシン


Vancomycin Hydrochloride contains not less than1000 mg (potency) per mg, calculated on the anhydrousbasis. The potency of Vancomycin Hydrochloride isexpressed as mass (potency) of vancomycin(C66H75Cl2N9O24: 1449.25).

Description Vancomycin Hydrochloride occurs as a whitepowder.

It is freely soluble in water, soluble in formamide, slightlysoluble in methanol, very slightly soluble in ethanol (95),and practically insoluble in acetonitrile.

It is hygroscopic.

Identiˆcation (1) Determine the absorption spectrum ofa solution of Vancomycin Hydrochloride (1 in 10,000) asdirected under the Ultraviolet-visible Spectrophotometry,and compare the spectrum with the Reference Spectrum orthe spectrum of a solution of Vancomycin HydrochlorideReference Standard prepared in the same manner as thesample solution: both spectra exhibit similar intensities ofabsorption at the same wavelengths.

(2) Determine the infrared absorption spectrum of Van-comycin Hydrochloride as directed in the potassiumbromide disk method under the Infrared Spectrophotomet-ry, and compare the spectrum with the Reference Spectrumor the spectrum of Vancomycin Hydrochloride ReferenceStandard: both spectra exhibit similar intensities of absorp-

tion at the same wave numbers.(3) Dissolve 20 mg of Vancomycin Hydrochloride in 10

mL of water, and add 1 drop of silver nitrate TS: a white tur-bidity is produced.

Optical rotation [a]D20: －30 –－409(0.2 g calculated onthe anhydrous basis, water, 20 mL, 100 mm).

pH The pH of a solution obtained by dissolving 0.25 g ofVancomycin Hydrochloride in 5 mL of water is between 2.5and 4.5.

Purity (1) Heavy metals—Proceed with 1.0 g of Van-comycin Hydrochloride according to Method 2, and per-form the test. Prepare the control solution with 2.0 mL ofStandard Lead Solution (not more than 20 ppm).

(2) Related substances—Dissolve 0.10 g of VancomycinHydrochloride in 10 mL of the mobile phase A, and use thissolution as the sample solution. Pipet 1 mL of the sample so-lution, add the mobile phase A to make exactly 25 mL, anduse this solution as the standard solution. Perform the testwith exactly 20 mL each of the sample solution and the stan-dard solution as directed under the Liquid Chromatographyaccording to the following conditions. If necessary, proceedwith 20 mL of the mobile phase A in the same manner tocompensate for the base line. Determine each peak area bythe automatic integration method: the area of each peakother than vancomycin from the sample solution is not morethan the peak area of vancomycin from the standard solu-tion, and the total area of the peaks other than vancomycinis not more than 3 times of the peak area of vancomycinfrom the standard solution.Operating conditions—




Mobile phase A: A mixture of triethylamine buŠer solu-tion, pH 3.2, acetonitrile and tetrahydrofuran (92:7:1). Ad-just the amount of acetonitrile so that the retention time ofvancomycin is 7.5 to 10.5 minutes.

Mobile phase B: A mixture of triethylamine buŠer solu-tion, pH 3.2, acetonitrile and tetrahydrofuran (70:29:1).



Mobile phaseA (z)

Mobile phaseB (z)

0 – 12 100 012 – 20 100→ 0 0→ 10020 – 22 0 100

Flow rate: 1.5 mL per minute.Time span of measurement: As long as about 2.5 times of


the retention time of vancomycin after the solvent peak.System suitability—

Test for required detectability: Conˆrm that the peak areaof vancomycin obtained from 20 mL of the standard solutionis equivalent to 3 to 5z of that from 20 mL of the sample so-lution.

System performance: Dissolve 5 mg of VancomycinHydrochloride in 10 mL of water, heat at 659C for 48 hours,and cool to the ordinal temperature. When the procedure isrun with 20 mL of this solution under the above operatingconditions, related substance 1, vancomycin and related sub-stance 2 are eluted in this order, the resolution between thepeaks of the related substance 1 and vancomycin is not lessthan 3, the theoretical plates of the peak of vancomycin isnot less than 1500 steps, and the related substance 2 is elutedbetween 15 and 18 minutes.

System repeatability: When the test is repeated 5 timeswith 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peakarea of vancomycin is not more than 2.0z.

Water Not more than 5.0z (0.1 g, volumetric titration,direct titration. Use a mixture of formamide for water deter-mination and methanol for water determination (3:1)).




for test organism [5] under (1) Agar media for seed and baselayer. Adjust the pH of the medium so that it will be 6.2 to

6.4 after sterilization.(3) Standard solutions—Weigh accurately an amount of

Vancomycin Hydrochloride Reference Standard, equivalentto about 25 mg (potency), dissolve in water to make exactly25 mL, and use this solution as the standard stock solution.Keep the standard stock solution at 59C or below, and usewithin 7 days. Take exactly a suitable amount of the stan-dard stock solution before use, add 0.1 mol/L phosphatebuŠer solution, pH 4.5 to make solutions so that each mLcontains 100 mg (potency) and 25 mg (potency), and use thesesolutions as the high concentration standard solution andthe low concentration standard solution, respectively.

(4) Sample solutions—Weigh accurately an amount ofVancomycin Hydrochloride, equivalent to about 25 mg(potency), and dissolve in water to make exactly 25 mL.Take exactly a suitable amount of this solution, add 0.1mol/L phosphate buŠer solution, pH 4.5 to make solutionsso that each mL contains 100 mg (potency) and 25 mg (poten-cy), and use these solutions as the high concentration samplesolution and the low concentration sample solution, respec-tively.


Xylitol Injectionキシリトール注射液



15411541

General Rulesfor Crude Drugs

Change the paragraph 1 to read:

1. Crude drugs in the monographs include medici-nal parts obtained from plants or animals, cell inclu-sions and secretes separated from the origins, their ex-tracts, and minerals. General Rules for Crude Drugsand the Crude Drugs in General Tests, Processes andApparatus are applicable to the followings:

Acacia, Achyranthes Root, Agar, Agar Powder,Akebia Stem, Alisma Rhizome, Aloe, Alpinia O‹cina-rum Rhizome, Amomum Seed, Anemarrhena Rhi-zome, Angelica Dahurica Root, Apricot Kernel, Are-ca, Artemisia Capillaris Flower, Asiasarum Root,Asparagus Tuber, Astragalus Root, Atractylodes Lan-cea Rhizome, Atractylodes Rhizome, Bear Bile, Bearb-erry Leaf, Belladonna Root, Benzoin, Bitter Carda-mon, Bitter Orange Peel, Bupleurum Root, BurdockFruit, Calumba, Capsicum, Cardamon, Cassia Seed,Catalpa Fruit, Chrysanthemum Flower, Chuling,Cimicifuga Rhizome, Cinnamon Bark, Citrus UnshiuPeel, Clematis Root, Clove, Cnidium Rhizome, CoixSeed, Condurango, Coptis Rhizome, Corn Starch,Cornus Fruit, Corydalis Tuber, Cyperus Rhizome,Digenea, Digitalis, Dioscorea Rhizome, EphedraHerb, Eucommia Bark, Evodia Fruit, Fennel, For-sythia Fruit, Fritillaria Bulb, Gambir, Gardenia Fruit,Gastrodia Tuber, Gentian, Geranium Herb, Ginger,Ginseng, Glehnia Root, Glycyrrhiza, Gypsum, HempFruit, Hoelen, Honey, Houttuynia Herb, ImmatureOrange, Imperata Rhizome, Ipecac, Japanese AngelicaRoot, Japanese Gentian, Japanese Valerian, Jujube,Jujube Seed, Lithospermum Root, Longgu, LoquatLeaf, Magnolia Bark, Magnolia Flower, MallotusBark, Mentha Herb, Moutan Bark, Mulberry Bark,Notopterygium Rhizome, Nuphar Rhizome, NuxVomica, Ophiopogon Tuber, Oriental Bezoar, OysterShell, Panax Rhizome, Peach Kernel, Peony Root,Perilla Herb, Pharbitis Seed, Phellodendron Bark,

Picrasma Wood, Pinellia Tuber, Plantago Herb, Plan-tago Seed, Platycodon Root, Polygala Root, Polygo-num Root, Potato Starch, Powdered Acacia, Pow-dered Alisma Rhizome, Powdered Aloe, PowderedAmomum Seed, Powdered Atractylodes Lancea Rhi-zome, Powdered Atractylodes Rhizome, PowderedCalumba, Powdered Capsicum, Powdered CinnamonBark, Powdered Clove, Powdered Cnidium Rhizome,Powdered Coix Seed, Powdered Coptis Rhizome,Powdered Cyperus Rhizome, Powdered Digitalis,Powdered Dioscorea Rhizome, Powdered Fennel,Powdered Gambir, Powdered Gardenia Fruit, Pow-dered Gentian, Powdered Geranium Herb, PowderedGinger, Powdered Ginseng, Powdered Glycyrrhiza,Powdered Hoelen, Powdered Ipecac, PowderedJapanese Angelica Root, Powdered Japanese Gentian,Powdered Japanese Valerian, Powdered MagnoliaBark, Powdered Moutan Bark, Powdered OysterShell, Powdered Panax Rhizome, Powdered PeachKernel, Powdered Peony Root, Powdered Phelloden-dron Bark, Powdered Picrasma Wood, PowderedPlatycodon Root, Powdered Polygala Root, PowderedPolypourus Sclerotium, Powdered Rhubarb, Pow-dered Rose Fruit, Powdered Scutellaria Root, Pow-dered Senega, Powdered Senna Leaf, Powdered Smi-lax Rhizome, Powdered Sophora Root, PowderedSweet Hydrangea Leaf, Powdered Swertia Herb, Pow-dered Tragacanth, Powdered Zanthoxylum Fruit,Prunella Spike, Pueraria Root, Red Ginseng, Rehman-nia Root, Rhubarb, Rice Starch, Rose Fruit, Rosin,SaŒower, SaŠron, Saposhnikovia Root, SaussureaRoot, Schisandra Fruit, Schizonepeta Spike, ScopoliaRhizome, Scutellaria Root, Senega, Senna Leaf,Sinomenium Stem, Smilax Rhizome, Sophora Root,Sweet Hydrangea Leaf, Swertia Herb, Toad Venom,Tragacanth, Trichosanthes Root, Uncaria Thorn,Wheat Starch, Zanthoxylum Fruit, Zedoary.

15431543

O‹cial Monographsfor Part II

Acaciaアラビアゴム


Identiˆcation To 1 g of powdered Acacia add 25 mL ofwater and 1 mL of sulfuric acid, and heat under a re‰ux con-denser in a boiling water bath for 60 minutes. After cooling,add gently 2.0 g of anhydrous sodium carbonate. To 1 mLof this solution add 9 mL of methanol, mix well, centrifuge,and use the supernatant liquid as the sample solution.Separately, dissolve 0.01 g of D-galactose in 1 mL water, addmethanol to make 10 mL, and use this solution as the stan-dard solution (1). Proceed with L-arabinose and with L-rhamnose monohydrate in the same manner as for thepreparation of the standard solution (1), and use so obtainedsolutions as the standard solution (2) and the standard solu-tion (3), respectively. Perform the test with these solutions asdirected under the Thin-layer chromatography. Spot 10 mLeach of the sample solution and the standard solutions (1),(2) and (3) on a plate of silica gel for thin-layer chro-matography. Develop the plate with a mixture of acetoneand water (9:1) to a distance of about 10 cm, and air-dry theplate. Spray evenly 1-naphthol-sulfuric acid TS on the plate,and heat at 1059C for 5 minutes: the three spots from thesample solution are the same with the spots of D-galactose,L-arabinose and L-rhamnose from the standard solution inthe color tone and the Rf value, respectively.


Purity (1) Insoluble residue—To 5.0 g of pulverized Aca-cia add 100 mL of water and 10 mL of dilute hydrochloricacid, and dissolve by gentle boiling for 15 minutes withswirling. Filter the warm mixture through a tared glass ˆlter(G3), wash the residue thoroughly with hot water, and dry at1059C for 5 hours: the mass of the residue does not exceed10.0 mg.

(2) Tannin-bearing gums—To 10 mL of a solution ofAcacia (1 in 50) add 3 drops of iron (III) chloride TS: nodark green color is produced.

(3) Glucose—Dissolve 0.01 g of glucose in 1 mL ofwater, add methanol to make 10 mL, and use this solution asthe standard solution. Proceed with the sample solution ob-tained in the Identiˆcation and the standard solution ob-tained here as directed in the Identiˆcation: any spot at theRf value corresponding to glucose from the standard solu-tion does not appear from the sample solution.

Powdered Acaciaアラビアゴム末


Identiˆcation To 1 g of Powdered Acacia add 25 mL ofwater and 1 mL of sulfuric acid, and heat under a re‰ux con-denser in a boiling water bath for 60 minutes. After cooling,add gently 2.0 g of anhydrous sodium carbonate. To 1 mLof this solution add 9 mL of methanol, mix well, centrifuge,and use the supernatant liquid as the sample solution.Separately, dissolve 0.01 g of D-galactose in 1 mL water, addmethanol to make 10 mL, and use this solution as the stan-dard solution (1). Proceed with L-arabinose and with L-rhamnose monohydrate in the same manner as for thepreparation of the standard solution (1), and use so obtainedsolutions as the standard solution (2) and the standard solu-tion (3), respectively. Perform the test with these solutions asdirected under the Thin-layer chromatography. Spot 10 mLeach of the sample solution and the standard solutions (1),(2) and (3) on a plate of silica gel for thin-layer chro-matography. Develop the plate with a mixture of acetoneand water (9:1) to a distance of about 10 cm, and air-dry theplate. Spray evenly 1-naphthol-sulfuric acid TS on the plate,and heat at 1059C for 5 minutes: the three spots from thesample solution are the same with the spots of D-galactose,L-arabinose and L-rhamnose from the standard solution inthe color tone and the Rf value, respectively.


Purity (1) Insoluble residue—To 5.0 g of Powdered Aca-cia add 100 mL of water and 10 mL of dilute hydrochloricacid, and dissolve by gentle boiling for 15 minutes withswirling. Filter the warm mixture through a tared glass ˆlter(G3), wash the residue thoroughly with hot water, and dry at1059C for 5 hours: the mass of the residue does not exceed10.0 mg.

(2) Tannin-bearing gums—To 10 mL of a solution ofPowdered Acacia (1 in 50) add 3 drops of iron (III) chlorideTS: no dark green color is produced.

(3) Glucose—Dissolve 0.01 g of glucose in 1 mL ofwater, add methanol to make 10 mL, and use this solution asthe standard solution. Proceed with the sample solution ob-tained in the Identiˆcation and the standard solution ob-tained here as directed in the Identiˆcation: any spot at theRf value corresponding to glucose from the standard solu-tion does not appear from the sample solution.

15441544 Supplement I, JP XIVO‹cial Monographs for Part II

Aloeアロエ


Aloe is the dried juice of the leaves mainly of Aloeferox Miller, or of hybrids of the species with Aloeafricana Miller or Aloe spicata Baker (Liliaceae).

It contains not less than 4.0z of barbaloin, calcu-lated on the basis of dried material.

Change the Loss on drying to read:

Loss on drying Not more than 12.0z.

Add the following next to Extract content:

Component determination Weigh accurately about 0.1 gof pulverized Aloe, add 40 mL of methanol, and heat undera re‰ex condenser on a water bath for 30 minutes. Aftercooling, ˆlter, and add methanol to the ˆltrate to make ex-actly 50 mL. Pipet 5 mL of the solution, add methanol tomake exactly 10 mL, and use this solution as the sample so-lution. Separately, weigh accurately about 10 mg of bar-baloin for component determination, previously dried in adesiccator (in vacuum, phosphorus (V) oxide) for 24 hours,add 0.04 g of oxalic acid dihydrate, and dissolve in methanolto make exactly 100 mL. Pipet 5 mL of the solution, addmethanol to make exactly 10 mL, and use this solution as thestandard solution. Perform the test with exactly 5 mL each ofthe sample solution and the standard solution as directed un-der the Liquid Chromatography according to the followingconditions, and measure the peak areas of barbaloin, AT andAS, of both solutions.

Amount (mg) of barbaloin＝WS×AT

AS× 2

WS: Amount (mg) of barbaloin for component determi-nation


length: 360 nm).Column: A stainless steel column about 6 mm in inside di-

ameter and about 15 cm in length, packed with octadecyl-silanized silica gel for liquid chromatography (5 mm in parti-cle diameter).



Flow rate: Adjust the ‰ow rate so that the retention timeof barbaloin is about 12 minutes.System suitability—

System performance: To about 10 mg of barbaloin forcomponent determination add 0.04 g of oxalic acid dihy-drate, and dissolve in methanol to make exactly 100 mL.Pipet 5 mL of the solution, add 1 mL of a solution of ethen-

zamide in methanol (1 in 2000) and methanol to make ex-actly 10 mL. When the procedure is run with 5 mL of this so-lution under the above operating conditions except thewavelength of 300 nm, barbaloin and ethenzamide are elut-ed in this order with the resolution between these peaksbeing not less than 2.0.

System repeatability: When the test is repeated 6 timeswith 5 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peakarea of barbaloin is not more than 1.5z.

Powdered Aloeアロエ末


Identiˆcation (2) To 0.2 g of Powdered Aloe add 10 mLof methanol, shake for 5 minutes, ˆlter, and use the ˆltrateas the sample solution. Separately, dissolve 1 mg of bar-baloin for thin-layer chromatography in 1 mL of methanol,and use this solution as the standard solution. Perform thetest with these solutions as directed under the Thin-layerChromatography. Spot 10 mL each of the sample solutionand the standard solution on a plate of silica gel for thin-lay-er chromatography. Develop the plate with a mixture ofethyl acetate, acetone, water and acetic acid (100) (20:5:2:2)to a distance of about 10 cm, and air-dry the plate. Examineunder ultraviolet light (main wavelength: 365 nm): one spotamong several spots from the sample solution has the samecolor tone and the same Rf value with the red ‰uorescentspot from the standard solution.

Add the following:

Alpinia O‹cinarum RhizomeAlpiniae o‹cinari Rhizoma

リョウキョウ

Alpinia O‹cinarum Rhizome is the rhizome of Al-pinia o‹cinarum Hance (Zingiberaceae).

Description Alpinia O‹cinarum Rhizome is a slightlycurved and cylindrical rhizome, sometimes branched; 2 to 8cm in length, 0.6 to 1.5 cm in diameter; externally red-brownto dark brown with ˆne striped lines, grayish white nodesand several traces of rootlet; hard to break; fracture surface,light brown in color and thickness of cortex is as same asthat of stele.

Odor, characteristic; taste, extremely pungent.Under a microscope, transverse section reveals epidermal

cells often containing resin-like substances; cortex, endoder-mis and stele present beneath the epidermis; cortex and steledivided by endodermis; vascular bundles surrounded byˆbers, scattered throughout the cortex and stele, cortex and

15451545Supplement I, JP XIV O‹cial Monographs for Part II

stele composed of parenchyma interspersed with oil cells;parenchymatous cells containing solitary crystals of calciumoxalate and starch grains, starch grains generally simple(sometimes 2- to 8-compound), ovate, oblong or narrowlyovate, 10 – 40 mm in diameter and with an eccentric navel.

Identiˆcation To 0.5 g of pulverized Alpinia O‹cinarumRhizome add 5 mL of acetone, shake for 5 minutes, andˆlter. Perform the test with the ˆltrate as directed under theThin-layer Chromatography. Spot 5 mL of the ˆltrate on aplate of silica gel for thin-layer chromatography, develop theplate with a mixture of cyclohexane, ethyl acetate and aceticacid (100) (12:8:1) to a distance of about 10 cm, and air-drythe plate: two yellow-brown spots appear at around Rf 0.4 –0.5.

Loss on drying Not more than 15.0z (6 hours).

Total ash Not more than 7.5z.

Acid-insoluble ash Not more than 1.5z.

Extract content Dilute ethanol-extract: not less than14.0z.

Arecaビンロウジ


Identiˆcation Weigh 3 g of pulverized Areca in a glass-stoppered centrifuge tube, and add 30 mL of diethyl etherand 5 mL of sodium hydroxide TS, stopper tightly, shakefor 5 minutes, centrifuge, and separate the diethyl etherlayer. Evaporate the diethyl ether on a water bath, dissolvethe residue in 1.5 mL of methanol, ˆlter, and use the ˆltrateas the sample solution. Separately, dissolve 5 mg of areco-line hydrobromide for thin-layer chromatography in 1 mLof methanol, and use this solution as the standard solution.Perform the test with these solutions as directed under theThin-layer chromatography. Spot 5 mL each of the samplesolution and the standard solution on a plate of silica gel forthin-layer chromatography. Develop the plate with a mix-ture of acetone, water and acetic acid (100) (10:6:1) to a dis-tance of about 10 cm, and air-dry the plate. Spray evenlyiodine TS on the plate: one spot among the spots from thesample solution and a red-brown spot from the standard so-lution show the same color tone and the same Rf value.

Asiasarum Rootサイシン


Description Asiasarum Root is a nearly cylindrical rhi-zome with numerous thin and long roots, externally lightbrown to dark brown. The root, about 15 cm in length,

about 0.1 cm in diameter, with shallow longitudinal wrinkleson the surface, and brittle. The rhizome, 2 – 4 cm in length,0.2 – 0.3 cm in diameter, often branched, with longitudinalwrinkles on the surface; internode short; each node hasseveral scars of petiole and peduncle, and several thin andlong roots.

Odor, characteristic; taste, acrid, with some sensation ofnumbness on the tongue.


Purity (1) Terrestrial part—Any terrestrial parts are notfound.


Purity(3) Aristolochic acid I—To exactly 2.0 g of pulverized

Asiasarum Root add exactly 50 mL of diluted methanol (3 in4), shake for 15 minutes, ˆlter, and use the ˆltrate as thesample solution. Separately, dissolve exactly 1.0 mg ofaristolochic acid I for crude drugs purity test in dilutedmethanol (3 in 4) to make exactly 100 mL. Pipet 1 mL of thissolution, add diluted methanol (3 in 4) to make exactly 25mL, and use this solution as the standard solution. Performthe test with exactly 20 mL each of the sample solution andthe standard solution as directed under the Liquid Chro-matography, according to the following conditions: the sam-ple solution shows no peak at the retention time corre-sponding to aristolochic acid I from the standard solution. Ifthe sample solution shows such a peak, repeat the test underdiŠerent conditions to conˆrm that the peak in question isnot aristolochic acid I.Operating conditions—

Detector: An ultraviolet or visible absorption photometer(wavelength: 400 nm).



Mobile phase: A mixture of a solution prepared by dis-solving 7.8 g of sodium dihydrogen phosphate dihydrate and2 mL of phosphoric acid in water to make 1000 mL andacetonitrile (11:9).

Flow rate: Adjust the ‰ow rate so that the retention timeof aristolochic acid I is about 15 minutes.System suitability—

Test for required detectability: Measure exactly 1 mL ofthe standard solution, and add diluted methanol (3 in 4) tomake exactly 10 mL. Conˆrm that the ratio, S/N, of the sig-nal (S) and noise (N) of aristolochic acid I obtained from 20mL of this solution is not less than 3. In this case, S meansthe peak height on the chromatogram not including noiseobtained by drawing an average line of the detector output,and N is 1/2 of the diŠerence between the maximum andminimum output signals of the baseline around the peak inthe range of 20 times the width at half-height of the peak.


System repeatability: When the test is repeated 6 timeswith 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peakarea of aristolochic acid I is not more than 5.0z.

Add the following:

Asparagus TuberAsparagi Tuber

テンモンドウ

Asparagus Tuber is the tuber of Asparagus cochin-chinensis Merrill (Liliaceae), from which most of thecork layer is removed, usually, after being steamed.

Description Asparagus Tuber is a fusiform to cylindricaltuber, 5 to 15 cm in length, 0.5 to 2 cm in diameter; external-ly light yellow-brown to light brown, translucent and oftenwith longitudinal wrinkles; ‰exible, or hard and easilybroken in texture; fractured surface, grayish yellow, glossyand horny.

Odor, characteristic; taste, sweet at ˆrst, followed by aslightly bitter aftertaste.

Under a microscope, a transverse section of AsparagusTuber reveals stone cells and bundles of them on outer layerof cortex; mucilaginous cells containing raphides of calciumoxalate in the parenchyma cells of cortex and centralcylinder; no starch grains.

Identiˆcation To 1 g of coarsely cut Asparagus Tuber add5 mL of a mixture of 1-butanol and water (40:7), shake for30 minutes, ˆlter, and use the ˆltrate as the sample solution.Perform the test with the sample solution as directed underthe Thin-layer Chromatography. Spot 10 mL of the samplesolution on a plate of silica gel for thin-layer chro-matography, develop the plate with a mixture of 1-butanol,water and acetic acid (100) (10:6:3) to a distance of about 10cm, and air-dry the plate. Spray evenly dilute sulfuric acidon the plate, and heat at 1059C for 2 minutes: the spot of ared-brown at ˆrst then changes to brown color appears ataround Rf 0.4.



Add the following:

Burdock FruitArctii Fructus

ゴボウシ

Burdock Fruit is the fruit of Arctium lappa Linn áe(Compositae).

Description Burdock Fruit is slightly curved, long obovateachene, 5 to 7 mm in length, 2.0 to 3.2 mm in width, 0.8 to1.5 mm in thickness; externally grayish brown to brown,with black spots; hollow about 1 mm in diameter at onebroad end; ‰at, indistinct, longitudinal ridge at the othernarrow end. 100 fruits weighing 1.0 to 1.5 g.

Practically odorless; taste, bitter and oily.Under a microscope, transverse section reveals an exocarp

of single-layered epidermal tissue, mesocarp of slightlyscleriˆed parenchyma, and endocarp of a single layer ofstone cells; seed coat composed of radially elongated, scleri-ˆed epidermis, and parenchyma several cells thick; paren-chymatous cells of the mesocarp contain a brown substance;stone cells of endocarp contain solitary, discrete crystals ofcalcium oxalate; cotyledons with starch grains, oil drops,aleurone grains, and minute crystals of calcium oxalate.

Identiˆcation To 0.5 g of pulverized Burdock Fruit add 20mL of methanol, shake for 10 minutes, ˆlter, and use ˆltrateas the sample solution. Perform the test with the sample so-lution as directed under the Thin-layer Chromatography.Spot 5 mL of the sample solution on a plate of silica gel forthin-layer chromatography, develop the plate with a mixtureof acetone, ethyl acetate and water (15:10:1) to a distance ofabout 10 cm, and air-dry the plate. Spray evenly dilute sul-furic acid on the plate, and heat at 1059C for 5 minutes: ared-purple spot appears at around Rf 0.4.





Capsicumトウガラシ

Delete the Extract content.

Powdered Capsicumトウガラシ末

Delete the Extract content.


Add the following:

Chrysanthemum FlowerChrysanthemi Flos

キクカ

Chrysanthemum Flower is the capitulum of 1)Chrysanthemum morifolium Ramatulle or 2)Chrysanthemum indicum Linn áe (Compositae).

Description 1) Chrysanthemum Flower is capitulum, 15to 40 mm in diameter; involucre consisting of 3 to 4 rows ofinvolucral scales; the outer involucral scale linear to lanceo-late, inner involucral scale narrow ovate to ovate; ligulate‰owers are numerous, white to yellow; tubular ‰owers insmall number, light yellow-brown; tubular ‰owers oc-casionally degenerate; outer surface of involucre green-brown to brown; light in texture and easy to break.

Odor, characteristic; taste, slightly bitter.2) Chrysanthemum Flower is capitulum, 3 to 10 mm in

diameter; involucre consisting of 3 to 5 rows of involucralscales; the outer involucral scale linear to lanceolatae, innerinvolucral scale narrow ovate to ovate; ligulate ‰ower is sin-gle, yellow to light yellow-brown; tubular ‰owers innumerous, light yellow-brown; outer surface of involucreyellow-brown to brown; light in texture and easy to break.

Odor, characteristic; taste, slightly bitter.

Identiˆcation To 1 g of pulverized Chrysanthemum Flow-er add 20 mL of methanol, shake for 10 minutes, and ˆlter.Evaporate the ˆltrate to dryness, dissolve the residue in 1 mLof methanol, and use this as the sample solution. Separately,dissolve 1 mg of luteolin for thin-layer chromatography in 1mL of methanol, and use this as the standard solution. Per-form the test with these solutions as directed under the Thin-layer Chromatography. Spot 10 mL each of the sample solu-tion and the standard solution on a plate of silica gel forthin-layer chromatography, develop the plate with a mixtureof ethyl acetate, 2-butanone, water and formic acid(25:3:1:1) to a distance of about 10 cm, and air-dry the plate.Spray evenly iron (III) chloride-methanol TS on the plate:one of several spots obtained from the sample solution hasthe same color tone and the same Rf value with the darkgreen spot obtained from the standard solution.





Cinnamon Barkケイヒ

Change the Total ash to read:


Powdered Cinnamon Barkケイヒ末



Add the following:

Clematis RootClematidis Radix

イレイセン

Clematis Root is the root and rhizome of Clematischinensis Osbeck, Clematis manshurica Ruprecht, orClematis hexapetala Pallas (Ranunculaceae).

Description Clematis Root consists of short rhizome andnumerous slender roots. The root, 10 to 20 cm in length, 1 to2 mm in diameter, externally brown to blackish brown, withˆne longitudinal wrinkles, brittle. The cortex easily separa-ble from central cylinder; root, grayish white to light brownin the transverse section, light grayish yellow to yellow in thecentral cylinder; under a magnifying glass, central cylinderalmost round, slight 2 to 4 sinuses on xylem. The rhizome, 2to 4 cm in length, 5 to 20 mm in diameter, externally lightgrayish brown to grayish brown; cortex peeled oŠ andˆbrous, often with rising node; apex having the residue ofligniˆed stem.

Odor, slight; practically tasteless.Under a microscope, transverse section of root reveals a

uni-layered epidermis in the outermost layer; with exodermislying just inside of the epidermis; cortex and stele divided byendodermis; cortex composed of parenchymatous tissue;xylem with 2 – 4 small concavities where phloem is present;parenchymatous cells contain both simple and 2- to 8-com-pound starch grains.

Identiˆcation (1) To 0.5 g of pulverized Clematis Rootadd 10 mL of water, and boil for 2 to 3 minutes. After cool-ing, shake vigorously: a lasting ˆne foam appears.

(2) To 0.5 g of pulverized Clematis Root add 3 mL ofacetic anhydride, warm on a water bath for 2 minutes, andˆlter. To the ˆltrate add 1 mL of sulfuric acid gently: abrown color appears at the zone of contact.






Cod Liver Oil肝油


Description Cod Liver Oil is a yellow to orange oily liquid.It has a characteristic, slightly ˆshy odor and a mild taste.

It is miscible with chloroform.It is slightly soluble in ethanol (95), and practically insolu-

ble in water.It is decomposed by air or by light.

Coptis Rhizomeオウレン


Identiˆcation(2) To 0.5 g of pulverized Coptis Rhizome add 20 mL of

methanol, shake for 2 minutes, ˆlter, and use the ˆltrate asthe sample solution. Separately, dissolve 1 mg of BerberineChloride Reference Standard in 1 mL of methanol, and usethis solution as the standard solution. Perform the test withthese solutions as directed under the Thin-layer Chro-matography. Spot 5 mL each of the sample solution and thestandard solution on a plate of silica gel for thin-layer chro-matography. Develop the chromatogram with a mixture of1-butanol, water and acetic acid (100) (7:2:1) to a distance ofabout 10 cm, and air-dry the plate. Examine under ultravio-let light (main wavelength: 365 nm): one spot among thespots from the sample solution and a spot from the standardsolution with yellow to yellow-green ‰uorescence show thesame color tone and the same Rf value.

Powdered Coptis Rhizomeオウレン末


Identiˆcation(2) To 0.5 g of Powdered Coptis Rhizome add 20 mL of

methanol, shake for 2 minutes, ˆlter, and use the ˆltrate asthe sample solution. Separately, dissolve 1 mg of BerberineChloride Reference Standard in 1 mL of methanol, and usethis solution as the standard solution. Perform the test withthese solutions as directed under the Thin-layer Chro-matography. Spot 5 mL each of the sample solution and the

standard solution on a plate of silica gel for thin-layer chro-matography. Develop the chromatogram with a mixture of1-butanol, water and acetic acid (100) (7:2:1) to a distance ofabout 10 cm, and air-dry the plate. Examine under ultravio-let light (main wavelength: 365 nm): one spot among thespots from the sample solution and a spot from the standardsolution with yellow to yellow-green ‰uorescence show thesame color tone and the same Rf value.

Add the following:

Eucommia BarkEucommiae Cortex

トチュウ

Eucommia Bark is the bark of Eucommia ulmoidesOliver (Eucommiaceae).

Description Eucommia Bark is a semi-tubular or plate-likebark, 2 to 6 mm in thickness; externally pale grayish brownto grayish brown, and rough in texture, sometimes reddish-brown due to the cork layer falling oŠ; internally dark vio-let, smooth and covered with a linear pattern that runs lon-gitudinally, silk-like threads of gutta-percha (a thermoplas-tic rubber-like substance) appearing when broken.

Odor faint but distinctive; taste slightly sweet.Under a microscope, transverse section reveals paren-

chymatous cells containing gutta-percha; phloem with stone-cell and ˆber layers; rays in rows of 2 – 3 cells; calcium oxa-late crystals absent.

Identiˆcation Put 1 g of pulverized Eucommia Bark in aglass-stoppered centrifuge tube, add 10 mL of water and20 mL of diethyl ether, shake for 15 minutes, and centrifuge.Take the diethyl ether layer so obtained, evaporate thediethyl ether on a water bath, and add 1 mL of ethanol(99.5) to the residue: colloidal substances appear.




Extract content Dilute ethanol-extract: not less than 7.0z.

Evodia Fruitゴシュユ


Evodia Fruit is the fruit of Evodia rutaecarpa Ben-tham or Evodia o‹cinalis Dode or Evodia bodinieriDode (Rutaceae).


Add the following:

Fritillaria BulbFritillariae Bulbus

バイモ

Fritillaria Bulb is the bulb of Fritillaria verticillataWilldenow var. thunbergii Baker (Liliaceae).

Description Fritillaria Bulb is a depressed spherical bulb, 2to 3 cm in diameter, 1 to 2 cm in height, consisting of 2thickened scaly leaves often separated; externally and inter-nally white to light yellow-brown in color; inside base is in aslightly dark color; the bulb sprinkled with lime beforedrying is dusted with white powder; fractured surface, whitein color and powdery.

Odor, slight and characteristic; taste, bitter.Under the microscope, a transverse section reveals the

outermost layer (epidermis) to be composed of a single layerof cells; numerous vascular bundles scattered throughout theparenchyma inside of the epidermis; parenchyma ˆlled withstarch grains; starch grains are mainly simple (rarely 2 – 3composite), 5 – 50 mm in diameter, narrowly ovate to ovateor triangular to obovate, stratiform ˆgure obvious; epider-mal cells and parenchymatous cells near the vessels containsolitary crystals of calcium oxalate.

Identiˆcation Put 2 g of pulverized Fritillaria Bulb in aglass-stoppered centrifuge tube, add 10 mL of ammonia TSand 20 mL of a mixture of ethyl acetate and diethyl ether(1:1), shake for 20 minutes, and centrifuge. Take the upperlayer, add 20 g of anhydrous sodium sulfate to the layer,shake, and ˆlter. Evaporate the ˆltrate to dryness, dissolvethe residue in 1 mL of ethanol (99.5), and use this solution asthe sample solution. Perform the test with the sample solu-tion as directed under the Thin-layer Chromatography. Spot10 mL of the sample solution on a plate of silica gel for thin-layer chromatography, develop the plate with a mixture ofethyl acetate, methanol and ammonia solution (28) (17:2:1)to a distance of about 10 cm, and air-dry the plate. Sprayevenly DragendorŠ's TS for spraying on the plate: two spotsof a yellow-red color appear at around Rf 0.4 and at aroundRf 0.6.





Gardenia Fruitサンシシ


Gardenia Fruit is the fruit of Gardenia jasminoidesEllis (Rubiaceae).

It contains not less than 3.0z of geniposide(C17H24O10: 388.37), calculated on the basis of driedmaterial.


Identiˆcation (1) To 1.0 g of pulverized Gardenia Fruit,previously dried in a desiccator (silica gel) for 24 hours, add100 mL of hot water, warm the mixture between 609C and709C for 30 minutes with frequent shaking, and ˆlter aftercooling. To 1.0 mL of the ˆltrate add water to make 10 mL:the color of the resulting solution is yellow and is not lighterthan that of the following control solution.

Control solution: Dissolve 9.8 mg of carbazochrome sodi-um sulfonate in water to make exactly 10 mL. Pipet 1 mL ofthis solution, and add water to make exactly 50 mL.


Loss on drying Not more than 13.0z.

Add the following next to Total ash:

Component determination Weigh accurately about 0.5 gof pulverized Gardenia Fruit, transfer into a glass-stopperedcentrifuge tube, add 40 mL of diluted methanol (1 in 2),shake for 15 minutes, centrifuge, and take the supernatantliquid. To the residue add 40 mL of diluted methanol (1 in2), and perform as the same as above. Combine the extractsso obtained, and add diluted methanol (1 in 2) to make ex-actly 100 mL. Pipet 5 mL of the solution, add methanol tomake exactly 20 mL, use this solution as the sample solu-tion. Separately, weigh accurately about 10 mg of genipo-side for component determination, previously dried in adesiccator (in vacuum, phosphorus (V) oxide) for 24 hours,and dissolve in methanol to make exactly 100 mL. Pipet 5mL of the solution, add methanol to make exactly 10 mL,and use this solution as the standard solution. Perform thetest with exactly 10 mL each of the sample solution and thestandard solution as directed under the Liquid Chro-matography according to the following conditions, andmeasure the peak areas of geniposide, AT and AS, of bothsolutions.

Amount (mg) of geniposide＝WS×AT

AS× 2

WS: Amount (mg) of geniposide for component determi-nation







of geniposide is about 15 minutes.System suitability—

System performance: Dissolve 1 mg each of geniposidefor component determination and caŠeine in methanol tomake 15 mL. When the procedure is run with 10 mL of thissolution under the above operating conditions, caŠeine andgeniposide are eluted in this order with the resolution be-tween these peaks being not less than 3.5.

System repeatability: When the test is repeated 6 timeswith 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peakarea of geniposide is not more than 1.5z.

Add the following:

Gastrodia TuberGastrodiae Tuber

テンマ

Gastrodia Tuber is the steamed tuber of Gastrodiaelata Blume (Orchidaceae).

Description Gastrodia Tuber is an irregularly curved and‰attened cylindrical to ‰attened fusiform tuber, 5 to 15 cmin length, 2 to 5 cm in diameter, 1 to 2 cm in thickness; exter-nally light yellow-brown to light yellowish white; with ringnodes, and irregular longitudinal wrinkles; hard in texture;fractured surface, dark brown to yellow-brown in color,with luster, horny and gluey.

Odor, characteristic; practically tasteless.Under a microscope, a transverse section reveals paren-

chyma cells containing needle raphides of calcium oxalate;starch grain absent.

Identiˆcation To 1 g of pulverized Gastrodia Tuber add 5mL of methanol, shake for 15 minutes, and ˆlter. Evaporatethe ˆltrate to dryness, dissolve the residue in 1 mL ofmethanol, and use this solution as the sample solution. Per-form the test with the sample solution as directed under theThin-layer Chromatography. Spot 10 mL of the sample solu-tion on a plate of silica gel for thin-layer chromatography,develop the plate with a mixture of ethyl acetate, methanoland water (8:2:1) to a distance of about 10 cm, and air-drythe plate. Spray evenly dilute sulfuric acid on the plate, andheat at 1059C for 1 minutes: a red-purple spot appears ataround Rf 0.4.



Extract content Dilute ethanol-soluble extract: not lessthan 16.0z.

Glycyrrhizaカンゾウ


Identiˆcation To 2 g of pulverized Glycyrrhiza add 10 mLof a mixture of ethanol (95) and water (7:3), heat by shakingon a water bath for 5 minutes, cool, ˆlter, and use the ˆltrateas the sample solution. Separately, dissolve 5 mg of Glycyr-rhizinic Acid Reference Standard in 1 mL of a mixture ofethanol (95) and water (7:3), and use this solution as thestandard solution. Perform the test with these solutions asdirected under the Thin-layer Chromatography. Spot 2 mLeach of the sample solution and the standard solution on aplate of silica gel with ‰uorescent indicator for thin-layerchromatography. Develop the plate with a mixture of 1-butanol, water and acetic acid (100) (7:2:1) to a distance ofabout 10 cm, and air-dry the plate. Examine under ultravio-let light (main wavelength: 254 nm): one spot among thespots from the sample solution and a dark purple spot fromthe standard solution show the same color tone and the sameRf value.

Powdered Glycyrrhizaカンゾウ末


Identiˆcation To 2 g of Powdered Glycyrrhiza add 10 mLof a mixture of ethanol (95) and water (7:3), heat by shakingon a water bath for 5 minutes, cool, ˆlter, and use the ˆltrateas the sample solution. Separately, dissolve 5 mg of Glycyr-rhizinic Acid Reference Standard in 1 mL of a mixture ofethanol (95) and water (7:3), and use this solution as thestandard solution. Perform the with these solutions test asdirected under the Thin-layer Chromatography. Spot 2 mLeach of the sample solution and the standard solution on aplate of silica gel with ‰uorescent indicator for thin-layerchromatography. Develop the plate with a mixture of 1-butanol, water and acetic acid (100) (7:2:1) to a distance ofabout 10 cm, and air-dry the plate. Examine under ultravio-let light (main wavelength: 254 nm): one spot among thespots from the sample solution and a dark purple spot fromthe standard solution show the same color tone and the sameRf value.


Add the following:

Hemp FruitCannabis Fructus

マシニン

Hemp Fruit is the fruit of Cannabis sativa Linn áe(Cannabidaceae).

Description Hemp Fruit is a slightly compressed void fruit,4 to 5 mm in length, 3 to 4 mm in diameter; externallygrayish green to grayish brown; pointed at one end, a scar ofgynophore at the other end, and crest lines on both sides;outer surface lustrous with white mesh-like pattern; slightlyhard pericarp; seed, slightly green in color and internally hasgrayish white albumen; 100 fruits weighing 1.6 to 2.7 g.

Practically odorless, aromatic on chewing; taste, mild andoily.

Under a microscope, a transverse section reveals the ex-ocarp to be a single-layered epidermis; mesocarp composedof parenchyma, a pigment cell layer and rows of short, smallcells; endocarp made up of a layer of radially elongatedstone cells; seed coat comprises a tubular cell layer andspongy tissue. Inside of the seed; exosperm consists of onelayer of parenchymatous cells, endosperm of one to severallayers of parenchymatous cells; most of the embryo com-posed of parenchyma, vascular bundles occurring in the cen-ter of hypocotyls and cotyledons; embryo parenchyma con-tains aleurone grains and oil drops.

Identiˆcation To 0.3 g of pulverized Hemp Fruit add 3 mLof methanol, shake for 10 minutes, centrifuge, and use thesupernatant liquid as the sample solution. Perform the testwith the sample solution as directed under the Thin-layerChromatography. Spot 5 mL of the sample solution on aplate of silica gel for thin-layer chromatography, develop theplate with a mixture of hexane and ethyl acetate (9:2) to adistance of about 10 cm, and air-dry the plate. Spray evenlyvanillin-sulfuric acid TS on the plate, and heat at 1059C for5 minutes: a dark blue-purple spot appears at around Rf 0.6.

Purity Bract—Hemp Fruit does not contain bract.




Imperata Rhizomeボウコン


Identiˆcation To 1 g of pulverized Imperata Rhizome add20 mL of hexane, allow the mixture to stand for 30 minuteswith occasional shaking, and ˆlter. Evaporate the hexane ofthe ˆltrate under reduced pressure, dissolve the residue in 5

mL of acetic anhydride, place 0.5 mL of this solution in atest tube, and add carefully 0.5 mL of sulfuric acid to maketwo layers: a red-brown color develops at the zone of con-tact, and the upper layer acquires a blue-green to blue-purplecolor.

Powdered Japanese Gentianリュウタン末


Identiˆcation To 0.5 g of Powdered Japanese Gentian add10 mL of methanol, shake for 20 minutes, ˆlter, and use theˆltrate as the sample solution. Separately, dissolve 1 mg ofgentiopicroside for thin-layer chromatography in 1 mL ofmethanol, and use this solution as the standard solution.Perform the test with these solutions as directed under theThin-layer Chromatography. Spot 10 mL each of the samplesolution and the standard solution on a plate of silica gelwith ‰uorescent indicator for thin-layer chromatography.Develop the plate with a mixture of ethyl acetate, ethanol(99.5) and water (8:2:1) to a distance of about 10 cm, andair-dry the plate. Examine under ultraviolet light (mainwavelength: 254 nm): one spot among the spots from thesample solution and a dark purple spot from the standardsolution show the same color tone and the same Rf value.

Add the following:

Jujube SeedZizyphi Semen

サンソウニン

Jujube Seed is the seed of Zizyphus jujuba Millervar. spinosa (Bunge) Hu ex H. F. Chou (Rham-naceae).

Description Jujube Seed is a compressed ovate to orbicu-lar, lenticular seed, 5 to 9 mm in lengh, 4 to 6 mm in width, 2to 3 mm in thickness, externally brown to dark red-brown,glossy; hilum at one end, charaza at the other end; seed coatsightly ˆexible, covering, milky white endosperm and lightyellow embryo. 100 seeds weighing 3.0 to 4.5 g.

Odor, slightly oily; taste, mild and slightly oily.Under a microscope, transverse section reveals seed coat

composed of an upper epidermis, parenchyma and lowerepidermis; upper epidermal cells scleriˆed and elongated inradial direction; lower epidermis covered with cuticle; en-dosperm composed of parenchyma, containing aggregatedcrystals of calcium oxalate, aleurone grains and starchgrains; cotyledons composed of parenchyma that containsaleurone grains, starch grains and oil drops.

Identiˆcation To 2 g of pulverized Jujube Seed add 10 mLof methanol, and heat under a re‰ux condenser for 10


minutes. After cooling, ˆlter, and use the ˆltrate as the sam-ple solution. Perform the test with the sample solution asdirected under the Thin-layer Chromatography. Spot 10 mLof the sample solution on a plate of silica gel with ‰uorescentindicator for thin-layer chromatography, develop the platewith a mixture of acetone, ethyl acetate, water and aceticacid (100) (10:10:3:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (mainwavelength: 254 nm): a purple spot appears at around Rf0.3, which shows a yellow-green to grayish green color afterspraying 1-naphthol-sulfuric acid TS on the plate and heat-ing at 1059C for 5 minutes.

Purity Foreign substance—Jujube Seed contains not morethan 1.0z of the endocarp and other foreign substances.




Add the following:

Loquat LeafEriobotryae Folium

ビワヨウ

Loquat Leaf is the leaf of Eriobotrya japonica Lin-dley (Rosaceae).

Description Loquat Leaf is an oblong to wide lanceolateleaf, 12 to 30 cm in length, 4 to 9 cm in width; pointed at theapex and wedge-shaped at the base; roughly serrate leaf withshort petiole; occasionally being cut into strips 5 to 10 mm inshorter diameter and several cm in longer diameter; uppersurface green to green-brown in color, lower surface lightgreen-brown with light brown woolly hairs; vein, light yel-low-brown in color, raised out on the lower surface of theleaf.

Odor, slight; practically tasteless.Under a microscope, a transverse section of Loquat Leaf

reveals thick cuticle on both surfaces; palisade tissue, mostly4 to 5 layers with several large cells without chloroplast; atmain vein, ring of collateral bundle partly cut by intrudingfundamental tissue at xylem side, and group of ˆber attach-ing to phloem; solitary and clustered crystals of calcium oxa-late in mesophyll; woolly hair, unicellular and curved, about25 mm in thickness, and up to 1.5 mm in length.

Identiˆcation To 0.3 g of pulverized Loquat Leaf add 10mL of methanol, warm on a water bath for 5 minutes withoccasional shaking, cool, ˆlter, and use the ˆltrate as thesample solution. Perform the test with this solution asdirected under the Thin-layer Chromatography. Spot 5 mLof the sample solution on a plate of silica gel for thin-layerchromatography. Develop the plate with a mixture ofacetonitrile and water (3:2) to a distance of about 10 cm, and

air-dry the plate. Spray evenly dilute sulfuric acid on theplate, and heat at 1059C for 10 minutes: a red-purple prin-cipal spot appears at around Rf 0.5.




Magnolia Barkコウボク

Change the Extract content to read:


Powdered Magnolia Barkコウボク末

Change the Extract content to read:


Add the following:

Magnolia FlowerMagnoliae Flos

シンイ

Magnolia Flower is the ‰ower bud of Magnoliasalicifolia Maximowicz, Magnolia kobus De Candolle,Magnolia biondii Pampanini, Magnolia sprengeriPampanini or Magnolia denudata Desrousseaux(Magnoliaceae).

Description Magnolia Flower is a fusiform ‰ower bud, 15to 45 mm in length, 6 to 20 mm in diameter of central part;often having ligneous peduncles on base; usually 3 bracts,externally with sparse hairs; brown to dark brown; or withdense hairs, grayish white to light yellow-brown, and the in-ner surface smooth and dark brown in color; interiorperianth of 9 pieces or 12 pieces, same size or exterior threepieces are smaller; 50 to 100 stamens and numerous pistils.Brittle in texture.

Odor, characteristic; taste, acrid and slightly bitter.

Identiˆcation To 1 g of pulverized Magnolia Flower add10 mL of methanol, shake for 15 minutes, ˆlter, and use theˆltrate as the sample solution. Perform the test with thesample solution as directed under the Thin-layer Chro-matography. Spot 20 mL of the sample solution on a plate of


silica gel for thin-layer chromatography, develop the platewith a mixture of ethyl acetate, acetone, water and formicacid (5:3:1:1) to a distance of about 10 cm, and air-dry theplate. Spray evenly DragendorŠ's TS for spraying on theplate: a yellow-red spot appears at around Rf 0.3.





Essential oil content Perform the test with 50.0 g of pul-verized Magnolia Flower as directed in the Essential oil con-tent under the Crude Drugs Test: the volume of essential oilis not less than 0.5 mL.

Morphine and Atropine Injectionモルヒネアトロピン注射液


Description Morphine and Atropine Injection is a clear,colorless liquid.

It is gradually colored by light.pH: 2.5 – 5.0


Identiˆcation To 2 mL of Morphine and Atropine Injec-tion add 2 mL of ammonia TS, and extract with 10 mL ofdiethyl ether. Filter the extract with a ˆlter paper, evaporatethe ˆltrate on a water bath to dryness, dissolve the residue in1 mL of ethanol (99.5), and use this solution as the samplesolution. Separately, dissolve 0.1 g of Morphine Hydrochlo-ride in 10 mL of water, perform with 2 mL of this solutionthe same procedure as used for preparation of the sample so-lution, and use the solution so obtained as the standard solu-tion (1). Separately, dissolve 3 mg of Atropine Sulfate Ref-erence Standard in 10 mL of water, perform with 2 mL ofthis solution the same procedure as used for preparation ofthe sample solution, and use the solution so obtained as thestandard solution (2). Perform the test with these solutionsas directed under the Thin-layer Chromatography. Spot 10mL each of the sample solution, the standard solution (1) andthe standard solution (2) on a plate of silica gel for thin-layerchromatography. Develop the plate with a mixture ofmethanol and ammonia solution (28) (200:3) to a distance ofabout 10 cm, and air-dry the plate. Spray evenly Dragendor-Š's TS on the plate: the two spots obtained from the samplesolution show the same color tone and the same Rf valuewith either spot of orange color obtained from the standardsolution (1) or the standard solution (2) (morphine and atro-pine).


Assay (1) Morphine hydrochloride—Pipet 2 mL of Mor-phine and Atropine Injection, add exactly 10 mL of the in-ternal standard solution, then add water to make 50 mL,and use this solution as the sample solution. Separately,weigh accurately about 25 mg of morphine hydrochloridefor assay, add exactly 10 mL of the internal standard solu-tion to dissolve, then add water to make 50 mL, and use thissolution as the standard solution. Perform the test with 20mL of the sample solution and the standard solution asdirected under the Liquid Chromatography according to thefollowing conditions, and calculate the ratios, QT and QS, ofthe peak area of morphine to that of the internal standard.


＝WS×QT

QS× 1.1679


Internal standard solution—A solution of Etilefrine Hydro-chloride (1 in 500).Operating conditions—




Mobile phase: Dissolve 1.0 g of sodium lauryl sulfate in500 mL of diluted phosphoric acid (1 in 1000), and adjustthe pH with sodium hydroxide TS to 3.0. To 240 mL of thissolution add 70 mL of tetrahydrofuran, and mix.


System performance: When the procedure is run with 20mL of the standard solution under the above operating con-ditions, morphine and the internal standard are eluted in thisorder with the resolution between these peaks being not lessthan 3.


(2) Atropine sulfate—Pipet 2 mL of Morphine andAtropine Injection, add exactly 2 mL of the internal stan-dard solution, and use this solution as the sample solution.Separately, weigh accurately about 15 mg of Atropine Sul-fate Reference Standard (separately determine its loss ondrying in the same manner as directed under Atropine Sul-fate), and dissolve in water to make exactly 50 mL. Pipet2 mL of this solution, add exactly 2 mL of the internal stan-dard solution, and use this solution as the standard solution.


Perform the test with 20 mL each of the sample solution andthe standard solution as directed under the Liquid Chro-matography according to the following conditions, and cal-culate the ratios, QT and QS, of the peak areas of atropine tothat of the internal standard.

Amount (mg) of atropine sulfate [(C17H23NO3)2.H2SO4.H2O]

＝WS×QT

QS×

125× 1.027

WS: Amount (mg) of Atropine Sulfate Reference Stand-ard, calculated on the dried basis

Internal standard solution—A solution of Etilefrine Hydro-chloride (1 in 12,500).Operating conditions—

Column, column temperature, and mobile phase: Proceedas directed in the operating conditions in the Assay (1).



System performance: When the procedure is run with20 mL of the sample solution under the above operating con-ditions, morphine, the internal standard and atropine areeluted in this order, and the resolution between morphineand the internal standard is not less than 3.

System repeatability: When the test is repeated 6 timeswith 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratiosof the peak area of atropine to that of the internal standardis not more than 1.0z.

Mulberry Barkソウハクヒ


Identiˆcation Heat 1 g of pulverized Mulberry Bark with20 mL of hexane under a re‰ux condenser on a water bathfor 15 minutes, and ˆlter. Evaporate the hexane of theˆltrate under reduced pressure, dissolve the residue in 10 mLof acetic anhydride, place 0.5 mL of the solution in a testtube, and add carefully 0.5 mL of sulfuric acid to make twolayers: a red-brown color develops at the zone of contact.

Add the following:

Notopterygium RhizomeNotopterygii Rhizoma

キョウカツ

Notopterygium Rhizome is the rhizome and root ofNotopterygium incisum Ting ex H. T. Chang orNotopterygium forbesii Boissieu (Umbelliferae).

Description Notopterygium Rhizome is slightly curved,cylindrical to conical, 3 to 10 cm in length, 5 to 20 mm in di-ameter; rhizome occasionally branched; externally yellow-brown to dark brown. The rhizome with nearly orbicular,hollowed stem scars at the apex, sometimes having shortresidue of stem; externally node rising, internode short; rootscars in warty processes on the node; externally root hascoarse longitudinal wrinkles and lateral root scars in wartyprocesses; light and slightly brittle in texture, easy to break.The transverse section of the rhizome reveals numerous radi-al cracks; cortex yellow-brown to brown; xylem light yellowto light grayish yellow; pith grayish white to light brown.Under a magnifying glass, the rhizome reveals brown, ˆnepoints of resin canals in the cortex and pith.

Odor, characteristic; taste, slightly acid at ˆrst, followedby a slightly pungent and slightly numbing aftertaste.

Under a microscope, transverse section shows the outer-most layer to be composed of a cork layer several to a dozenor so cells thick; collenchyma just inside of the cork layer;oil canals scattered in cortex, large ones more than 300 mm indiameter; intercellular space occurring in radial direction incortex; oil canals scattered in pith, large ones more than500 mm in diameter; parenchymatous cells contain simpleand 2- to 3-compound starch grains.

Identiˆcation To 0.3 g of pulverized Notopterygium Rhi-zome add 3 mL of hexane in a glass-stoppered centrifugetube, shake for 10 minutes, centrifuge, and use the super-natant liquid as the sample solution. Perform the test withthe sample solution as directed under the Thin-layer Chro-matography. Spot 10 mL of the sample solution on a plate ofoctadecylsilanized silica gel with ‰uorescent indicator forthin-layer chromatography, develop the plate with a mixtureof methanol and water (9:1) to a distance of about 10 cm,and air-dry the plate. Examine under ultraviolet light (mainwavelength: 365 nm): a bluish white ‰uorescent spot appearsat around Rf 0.5, which shows a dark purple color underultraviolet light (main wavelength: 254 nm).






Opium Alkaloids Hydrochlorides塩酸アヘンアルカロイド


Description Opium Alkaloids Hydrochlorides occur as awhite to light brown powder.

It is soluble in water, and slightly soluble in ethanol (95).It is colored by light.


Purity (1) Clarity and color of solution－Dissolve 0.5 gof Opium Alkaloids Hydrochlorides in 10 mL of water: thesolution is clear, and its absorbance at 420 nm is not morethan 0.20.

Powdered Panax Rhizomeチクセツニンジン末


Identiˆcation Shake 0.5 g of Powdered Panax Rhizomewith 10 mL of methanol for 10 minutes, ˆlter, and use theˆltrate as the sample solution. Separately, dissolve 2 mg ofchikusetsusaponin IV for thin-layer chromatography in1 mL of methanol, and use this solution as the standard so-lution. Perform the test with these solutions as directed un-der the Thin-layer Chromatography. Spot 5 mL each of thesample solution and the standard solution on a plate of silicagel for thin-layer chromatography. Develop the plate with amixture of ethyl acetate, water and formic acid (5:1:1) to adistance of about 10 cm, and air-dry the plate. Spray evenlydilute sulfuric acid on the plate, and heat the plate at 1109Cfor 5 minutes: one of several spots obtained from the samplesolution shows the same color tone and the same Rf valuewith the purple-red spot from the standard solution.

Peony Rootシャクヤク


Identiˆcation(2) To 2 g of pulverized Peony Root add 10 mL of

methanol, warm on a water bath for 5 minutes, cool, ˆlter,and use the ˆltrate as the sample solution. Separately, dis-solve 1 mg of Paeoni‰orin Reference Standard in 1 mL ofmethanol, and use this solution as the standard solution.Perform the test with these solutions as directed under theThin-layer Chromatography. Spot 10 mL each of the samplesolution and the standard solution on a plate of silica gel forthin-layer chromatography. Develop the plate with a mix-ture of acetone, ethyl acetate and acetic acid (100) (10:10:1)to a distance of about 10 cm, and air-dry the plate. Spray

evenly 4-methoxybenzaldehyde-sulfuric acid TS upon theplate, and heat at 1059C for 5 minutes: one spot among thespots from the sample solution and the purple-red spot fromthe standard solution show the same color tone and the sameRf value.


Assay Weigh accurately about 0.5 g of pulverized PeonyRoot, add 50 mL of diluted methanol (1 in 2), heat under are‰ux condenser on a water bath for 30 minutes, cool, andˆlter. To the residue add 50 mL of diluted methanol (1 in 2),and proceed in the same manner. Combine the ˆltrates, adddiluted methanol (1 in 2) to make exactly 100 mL, and usethis solution as the sample solution. Separately, weigh ac-curately about 10 mg of Paeoni‰orin Reference Standard(separately detarmine the water content), dissolve in dilutedmethanol (1 in 2) to make exactly 100 mL, and use this solu-tion as the standard solution. Perform the test with exactly10 mL each of the sample solution and the standard solutionas directed under the Liquid Chromatography according tothe following conditions. Determine the peak areas, AT andAS, of paeoni‰orin in each solution.

Amount (mg) of paeoni‰orin (C23H28O11)＝WS×AT

AS

WS: Amount (mg) of Paeoni‰orin Reference Standard,calculated on the anhydrous basis





Mobile phase: A mixture of water, acetonitrile and phos-phoric acid (850:150:1).

Flow rate: Adjust the ‰ow rate so that the retention timeof paeoni‰orin is about 10 minutes.System suitability—

System performance: Dissolve 1 mg each of Paeoni‰orinReference Standard and albi‰orin in diluted methanol (1 in2) to make 10 mL. When the procedure is run with 10 mL ofthis solution under the above operating conditions, albi‰orinand paeoni‰orin are eluted in this order with the resolutionbetween these peaks being not less than 2.5.

System repeatability: When the test is repeated 6 timeswith the standard solution under the above operating condi-tions, the relative standard deviation of the peak area ofpaeoni‰orin is not more than 1.5z.


Powdered Peony Rootシャクヤク末


Identiˆcation(2) To 2 g of Powdered Peony Root add 10 mL of

methanol, warm on a water bath for 5 minutes, cool, ˆlter,and use the ˆltrate as the sample solution. Separately, dis-solve 1 mg of Paeoni‰orin Reference Standard in 1 mL ofmethanol, and use this solution as the standard solution.Perform the test with these solutions as directed under theThin-layer Chromatography. Spot 10 mL each of the samplesolution and the standard solution on a plate of silica gel forthin-layer chromatography. Develop the plate with a mix-ture of acetone, ethyl acetate and acetic acid (100) (10:10:1)to a distance of about 10 cm, and air-dry the plate. Sprayevenly 4-methoxybenzaldehyde-sulfuric acid TS on theplate, and heat at 1059C for 5 minutes: one spot among thespots from the sample solution and the purple spot from thestandard solution show the same color tone and the same Rfvalue.

Phellodendron Barkオウバク


Identiˆcation(2) Use the ˆltrate obtained in (1) as the sample solution.

Separately, dissolve 1 mg of Berberine Chloride ReferenceStandard in 1 mL of methanol, and use this solution as thestandard solution. Perform the test with these solutions asdirected under the Thin-layer Chromatography. Spot 5 mLeach of the sample solution and the standard solution on aplate of silica gel for thin-layer chromatography. Developthe plate with a mixture of 1-butanol, water and acetic acid(100) (7:2:1) to a distance of about 10 cm, and air-dry theplate. Examine under ultraviolet light (main wavelength: 365nm): one spot among the spots from the sample solution anda spot with yellow to yellow-green ‰uorescence from thestandard solution show the same color tone and the same Rfvalue.

Powdered Phellodendron Barkオウバク末


Identiˆcation(2) Use the ˆltrate obtained in (1) as the sample solution.

Separately, dissolve 1 mg of Berberine Chloride ReferenceStandard in 1 mL of methanol, and use this solution as thestandard solution. Perform the test with these solutions as

directed under the Thin-layer Chromatography. Spot 5 mLeach of the sample solution and the standard solution on aplate of silica gel for thin-layer chromatography. Developthe plate with a mixture of 1-butanol, water and acetic acid(100) (7:2:1) to a distance of about 10 cm, and air-dry theplate. Examine under ultraviolet light (main wavelength: 365nm): one spot among the spots from the sample solution anda spot with yellow to yellow-green ‰uorescence from thestandard solution show the same color tone and the same Rfvalue.

Compound Phellodendron Powderfor Cataplasmパップ用複方オウバク散


Identiˆcation Shake thoroughly 0.2 g of Compound Phel-lodendron Powder for Cataplasm with 5 mL of methanol,ˆlter, and use the ˆltrate as the sample solution. Dissolve 1mg of Berberine Chloride Reference Standard in 1 mL ofmethanol, and use the solution as the standard solution. Per-form the test with these solutions as directed under the Thin-layer Chromatography. Spot 5 mL each of the sample solu-tion and the standard solution on a plate of silica gel forthin-layer chromatography. Develop the plate with a mix-ture of 1-butanol, water and acetic acid (100) (7:2:1) to a dis-tance of about 10 cm, air-dry the plate. Examine underultraviolet light (main wavelength: 365 nm): one spot amongthe spots from the sample solution and a spot with yellow toyellow-green ‰uorescence from the standard solution showthe same color tone and the same Rf value (berberine).

Phellodendron, Albumin Tannateand Bismuth Subnitrate Powderオウバクタンナルビンビスマス散


Identiˆcation (1) Shake thoroughly 0.1 g of Phelloden-dron, Albumin Tannate and Bismuth Subnitrate Powderwith 5 mL of methanol, ˆlter, and use the ˆltrate as the sam-ple solution. Dissolve 1 mg of Berberine Chloride ReferenceStandard in 1 mL of methanol, and use this solution as thestandard solution. Perform the test with these solutions asdirected under the Thin-layer Chromatography. Spot 5 mLeach of the sample solution and the standard solution on aplate of silica gel for thin-layer chromatography. Developthe plate with a mixture of 1-butanol, water and acetic acid(100) (7:2:1) to a distance of about 10 cm, air-dry the plate.Examine under ultraviolet light (main wavelength: 365 nm):one spot among the spots from the sample solution and aspot with yellow to yellow-green ‰uorescence from the stan-dard solution show the same color tone and the same Rf


value (berberine).


Phenovalin and Magnesium OxidePowderフェノバリンマグネシア散

Add the following:

Polygonum RootPolygoni Multi‰ori Radix

カシュウ

Polygonum Root is the root of Polygonum multi‰o-rum Thunberg (Polygonaceae), often being cut intoround slices.

Description Polygonum Root is nearly fusiform, 10 to 15cm in length, 2 to 5 cm in diameter; externally red-brown todark brown; roughly wrinkled; a cross section light red-brown or light grayish brown, with numerous abnormal vas-cular bundles scattering irregularly around the large vascularbundles near center; heavy and hard in texture.

Odor, slight and characteristic; taste, astringent andslightly bitter.

Under a microscope, transverse section reveals the outer-most layer to be several cells thick and composed of cork;cork cells contain brown substances; cortex composed ofparenchyma; abnormal vascular bundles, exhibiting a ringof cambium; xylem lies inside of the cambium, and phloemoutside; ˆbers lie outside the phloem; central portion of rootligniˆed; parenchymatous cells contain aggregated crystalsof calcium oxalate, and both simple and 2- to 8-compoundstarch grains; navel of starch grain obvious.

Identiˆcation To 1 g of pulverized Polygonum Root add10 mL of methanol, shake for 15 minutes, and ˆlter.Evaporate the ˆltrate to dryness, dissolve the residue in 2 mLof methanol, and use this as the sample solution. Performthe test with the sample solution as directed under the Thin-layer Chromatography. Spot 5 mL of the sample solution ona plate of silica gel for thin-layer chromatography, developthe plate with a mixture of ethyl acetate, water, methanoland acetic acid (100) (200:10:10:3) to a distance of about 10cm, and air-dry the plate. Examine under ultraviolet light(main wavelength: 365 nm): a ‰uorescent bluish white spotappears at around Rf 0.3.



Extract content Dilute ethanol-extract: not less than

17.0z.

Pueraria Rootカッコン

Change the originWlimits of content to read:

Pueraria Root is the root of Pueraria lobata Ohwi(Leguminosae), from which periderm has been re-moved.

It contains not less than 2.0z of puerarin (C21H20O9:416.38), calculated on the basis of dried material.


Identiˆcation To 2 g of pulverized Pueraria Root add 10mL of methanol, shake for 3 minutes, ˆlter, and use theˆltrate as the sample solution. Separately, dissolve 1 mg ofPuerarin Reference Standard in 1 mL of methanol, and usethis solution as the standard solution. Perform the test withthese solutions as directed under the Thin-layer Chro-matography. Spot 2 mL each of the sample solution and thestandard solution on a plate of silica gel for thin-layer chro-matography. Develop the plate with a mixture of ethylacetate, methanol and water (12:2:1) to a distance of about10 cm, and air-dry the plate. Examine under ultraviolet light(main wavelength: 365 nm): one of the spots from the sam-ple solution and a blue-white spot from the standard solu-tion show the same color tone and the same Rf value.

Add the following next to Total ash:

Assay Weigh accurately about 0.3 g of pulverized PuerariaRoot, add 50 mL of diluted methanol (1 in 2), and heat un-der a re‰ex condenser on a water bath for 30 minutes, cool,and ˆlter. To the residue add 50 mL of diluted methanol (1in 2), and perform as the same as above. Combine theˆltrates, add diluted methanol (1 in 2) to make exactly 100mL, and use this solution as the sample solution. Separately,weigh accurately about 10 mg of Puerarin Reference Stan-dard (separately detarmine the water content), add dilutedmethanol (1 in 2) to make exactly 100 mL, and use this solu-tion as the standard solution. Perform the test with 10 mLeach of the sample solution and the standard solution asdirected under the Liquid Chromatography according to thefollowing conditions, and measure the peak areas of puera-rin, AT and AS, of each solution.

Amount (mg) of puerarin (C21H20O9)＝WS×AT

AS

WS: Amount (mg) of Puerarin Reference Standard, calcu-lated on the anhydrous basis

Operating conditions－Detector: An ultraviolet absorption photometer (wave-





Mobile phase: A mixture of 0.05 molWL sodium dihydro-gen phosphate TS and acetonitrile (9:1).

Flow rate: Adjust the ‰ow rate so that the retention timeof puerarin is about 15 minutes.System suitability－

System performance: When the procedure is run with10 mL of the standard solution under the above operatingconditions, the number of theoretical plates and the symmet-ry coe‹cient of the peak of puerarin are not less than 3000and not more than 2.0, respectively.

System repeatability: When the test is repeated 6 timeswith 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peakarea of puerarin is not more than 1.5z.

Rhubarbダイオウ


Identiˆcation To 2 g of pulverized Rhubarb add 40 mL ofa mixture of tetrahydrofuran and water (7:3), shake for 30minutes, and centrifuge. Transfer the supernatant liquid to aseparator, add 13 g of sodium chloride, and shake for 30minutes. Separate the water layer with undissolved sodiumchloride, and adjust the pH to 1.5 by adding 1 molWLhydrochloric acid TS. Transfer this solution to another sepa-rator, add 30 mL of tetrahydrofuran, shake for 10 minutes,separate the tetrahydrofuran layer, and use this solution asthe sample solution. Separately, dissolve 1 mg of SennosideA Reference Standard in 4 mL of a mixture of tetrahydrofu-ran and water (7:3), and use this solution as the standard so-lution. Perform the test with these solutions as directed un-der the Thin-layer Chromatography. Spot 40 mL each of thesample solution and the standard solution on a plate of silicagel for thin-layer chromatography at 10 mm along the initialline. Develop the plate with a mixture of ethyl acetate, 1-propanol, water and acetic acid (100) (40:40:30:1) to a dis-tance of about 15 cm, and air-dry the plate. Examine underultraviolet light (main wavelength: 365 nm): one of the spotsfrom the sample solution and a red ‰uorescent spot from thestandard solution show the same color tone and the same Rfvalue.

Delete the Component determination and addthe following:

Assay Weigh accurately about 0.5 g of pulverizedRhubarb, add exactly 50 mL of a solution of sodium hydro-gen carbonate (1 in 1000), shake for 30 minutes, ˆlter, anduse the ˆltrate as the sample solution. Separately, weigh ac-curately about 10 mg of Sennoside A Reference Standard,

dissolve in a solution of sodium hydrogen carbonate (1 in1000) to make exactly 50 mL. Pipet 5 mL of this solution,add a solution of sodium hydrogen carbonate (1 in 1000) tomake exactly 20 mL and use this solution as the standard so-lution. Perform the test with exactly 10 mL of the sample so-lution and the standard solution as directed under the LiquidChromatography according to the following conditions, anddetermine the peak areas, AT and AS, of sennoside A in eachsolution.

Amount (mg) of sennoside A (C42H38O20)

＝WS×AT

AS×

14

WS: Amount (mg) of Sennoside A Reference Standard,calculated on the anhydrous basis


length: 340 nm).Column: A stainless steel column about 4 to 6 mm in in-

side diameter and about 15 to 25 cm in length, packed withoctadecylsilanized silica gel for liquid chromatography (5 to10 mm in particle diameter).


Mobile phase: A mixture of diluted acetic acid (100) (1 in80) and acetonitrile (4:1).

Flow rate: Adjust the ‰ow rate so that the retention timeof sennoside A is about 15 minutes.System suitability－

System performance: Dissolve 1 mg each of Sennoside AReference Standard and naringin for thin-layer chro-matography in a solution of sodium hydrogen carbonate (1in 1000) to make 10 mL. When the procedure is run with 20mL of this solution under the above operating conditions,sennoside A and naringin are eluted in this order with theresolution between these peaks being not less than 3.

System repeatability: When the test is repeated 6 timeswith 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peakarea of sennoside A is not more than 1.5z.

Powdered Rhubarbダイオウ末


Identiˆcation To 2 g of Powdered Rhubarb add 40 mL ofa mixture of tetrahydrofuran and water (7:3), shake for 30minutes, and centrifuge. Transfer the supernatant liquid to aseparator, add 13 g of sodium chloride, and shake for 30minutes. Separate the water layer with undissolved sodiumchloride, and adjust the pH to 1.5 with 1 molWL hydrochlor-ic acid TS. Transfer this solution to another separator, add30 mL of tetrahydrofuran, shake for 10 minutes, separatethe tetrahydrofuran layer, and use this solution as the sam-ple solution. Separately, dissolve 1 mg of Sennoside A


Reference Standarad in 4 mL of a mixture of tetrahydrofu-ran and water (7:3), and use this solution as the standard so-lution. Perform the test with these solutions as directed un-der the Thin-layer Chromatography. Spot 40 mL each of thesample solution and the standard solution on a plate of silicagel for thin-layer chromatography at 10 mm along the initialline. Develop the plate with a mixture of 1-propanol, ethylacetate, water and acetic acid (100) (40:40:30:1) to a distanceof about 15 cm, and air-dry the plate. Examine under ultrav-iolet light (main wavelength: 365 nm): one of the spots fromthe sample solution and a red ‰uorescent spot from the stan-dard solution show the same color tone and the same Rfvalue.


Assay Weigh accurately about 0.5 g of PowderedRhubarb, add exactly 50 mL of a solution of sodium hydro-gen carbonate (1 in 1000), shake for 30 minutes, ˆlter, anduse the ˆltrate as the sample solution. Separately, weigh ac-curately about 10 mg of Sennoside A Reference Standard,dissolve in a solution of sodium hydrogen carbonate (1 in1000) to make exactly 50 mL. Pipet 5 mL of this solution,add a solution of sodium hydrogen carbonate (1 in 1000) tomake exactly 20 mL, and use this solution as the standardsolution. Perform the test with exactly 10 mL each of thesample solution and the standard solution as directed underthe Liquid Chromatography according to the following con-ditions, and determine the peak areas, AT and AS, of senno-side A in each solution.


＝WS×AT

AS×

14

WS: Amount (mg) of Sennoside A Reference Standard,calculated on the anhydrous basis


length: 340 nm).Column: A stainless steel column about 4 to 6 mm in in-

side diameter and about 15 to 25 cm in length, packed withoctadecylsilanized silica gel for liquid chromatography (5 to10 mm in particle diameter).


Mobile phase: A mixture of diluted acetic acid (100) (1 in80) and acetonitrile (4:1).


System performance: Dissolve 1 mg each of Sennoside AReference Standard and naringin for thin-layer chro-matography in a solution of sodium hydrogen carbonate (1in 1000) to make 10 mL. When the procedure is run with 20mL of this solution under the above operating conditions,sennoside A and naringin are eluted in this order with theresolution between these peaks being not less than 3.


Scopolia Extract and EthylAminobenzoate Powderロートエキスアネスタミン散


Identiˆcation(4) Place 30 g of Scopolia Extract and Ethyl Aminoben-

zoate Powder in a glass-stoppered conical ‰ask, add 100 mLof water, shake for 30 minutes, and ˆlter immediately bysuction through a glass ˆlter (G3). Transfer the residue in the‰ask to the same glass ˆlter with the ˆltrate, and ˆlter theresidue by suction while pressing vigorously the residue onthe same glass ˆlter. Place 75 mL of the ˆltrate in a 300-mLbeaker, and add cautiously 10 mL of diluted sulfuric acid (1in 3). Add 0.2 mL of bromocresol green TS to this solution,and add dilute sulfuric acid dropwise while shaking thor-oughly, until the color of the solution changes from green toyellow-green. After cooling, place this solution in a separa-tor, wash with two 25-mL portions of a mixture of hexaneand diethyl ether (1:1) by shaking well, and place the waterlayer in another separator. Make slightly alkaline with am-monia TS, add immediately 30 mL of diethyl ether, andshake well. Wash the diethyl ether layer with two 10-mL por-tions of a saturated solution of sodium chloride, separate thediethyl ether layer, add 3 g of anhydrous sodium sulfate,shake, and ˆlter through a pledget of cotton. Evaporate theˆltrate to dryness, dissolve the residue in 0.2 mL of ethanol(95), and use this solution as the sample solution. Separate-ly, dissolve 2 mg of Atropine Sulfate Reference Standardand 1 mg of Scopolamine Hydrobromide Reference Stan-dard in 1 mL each of ethanol (95), and use these solutions asstandard solution (1) and standard solution (2). Perform thetest with these solutions as directed under the Thin-layerChromatography. Spot 10 mL each of the sample solutionand the standard solutions on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture ofacetone, water and ammonia solution (28) (90:7:3) to a dis-tance of about 10 cm, and dry the plate at 809C for 10minutes. After cooling, spray evenly DragendorŠ's TS forspraying on the plate: two principal spots from the samplesolution show the same color tone and the same Rf valuewith each yellow-red spot from the standard solutions,respectively.



Compound Scopolia Extract andTannic Acid Ointment複方ロートエキスタンニン軟膏

Compound Scopolia Extract andTannic Acid Suppositories複方ロートエキスタンニン坐剤

Scutellaria Rootオウゴン


Identiˆcation(2) To 2 g of pulverized Scutellaria Root add 10 mL of

methanol, warm on a water bath for 3 minutes, cool, ˆlter,and use the ˆltrate as the sample solution. Separately, dis-solve 1 mg of Baicalin Reference Standard in 1 mL ofmethanol, and use this solution as the standard solution.Perform the test with these solutions as directed under theThin-layer Chromatography. Spot 5 mL each of the samplesolution and the standard solution on a plate of silica gel forthin-layer chromatography. Develop the plate with a mix-ture of 1-butanol, water and acetic acid (100) (4:2:1) to a dis-tance of about 10 cm, and air-dry the plate. Spray evenly asolution of iron (III) chloride hexahydrate in methanol (1 in100) on the plate: one spot among the spots from the samplesolution and a dark green spot from the standard solutionshow the same color tone and the same Rf value.

Powdered Scutellaria Rootオウゴン末


Identiˆcation(2) To 2 g of Powdered Scutellaria Root add 10 mL of

methanol, warm on a water bath for 3 minutes, cool, ˆlter,and use the ˆltrate as the sample solution. Separately, dis-solve 1 mg of Baicalin Reference Standard in 1 mL ofmethanol, and use this solution as the standard solution.Perform the test with these solutions as directed under theThin-layer Chromatography. Spot 5 mL each of the samplesolution and the standard solution on a plate of silica gel forthin-layer chromatography. Develop the plate with a mix-ture of 1-butanol, water and acetic acid (100) (4:2:1) to a dis-tance of about 10 cm, and air-dry the plate. Spray evenly asolution of iron (III) chloride hexahydrate in methanol (1 in100) on the plate: one spot among the spots from the sample

solution and dark green spot from the standard solutionshow the same color tone and the same Rf value.

Senna Leafセンナ


Senna Leaf is the lea‰ets of Cassia angustifolia Vahlor Cassia acutifolia Delile (Leguminosae).

It contains not less than 1.0z of total sennosides[sennoside A (C42H38O20: 862.75) and sennoside B(C42H38O20: 862.75)], calculated on the basis of driedmaterial.


Identiˆcation(2) To 2 g of pulverized Senna Leaf add 40 mL of a mix-

ture of tetrahydrofuran and water (7:3), shake for 30minutes, and centrifuge. Transfer the supernatant liquid to aseparator, add 13 g of sodium chloride, and shake for 30minutes. Separate the water layer with undissolved sodiumchloride, and adjust the pH to 1.5 by adding 1 molWLhydrochloric acid TS. Transfer this solution to another sepa-rator, shake with 30 mL of tetrahydrofuran for 10 minutes,separate the tetrahydrofuran layer, and use this solution asthe sample solution. Separately, dissolve 1 mg of SennosideA Reference Standarad in 1 mL of a mixture of tetra-hydrofuran and water (7:3), and use this solution as the stan-dard solution. Perform the test as directed under the Thin-layer Chromatography with the sample solution and thestandard solution. Spot 10 mL each of these solutions on aplate of silica gel for thin-layer chromatography. Developthe plate with a mixture of 1-propanol, ethyl acetate, waterand acetic acid (100) (40:40:30:1) to a distance of about 15cm, and air-dry the plate. Examine under ultraviolet light(main wavelength: 365 nm): one spot among the spots fromthe sample solution and a red ‰uorescent spot from the stan-dard solution show the same color tone and the same Rfvalue.


Assay Weigh accurately about 0.5 g of pulverized SennaLeaf in a glass-stoppered centrifuge tube, add 25 mL ofdiluted methanol (7 in 10), shake for 30 minutes, centrifuge,and separate the supernatant liquid. To the residue adddiluted methanol (7 in 10) twice with a 10-mL portion, shakefor 10 minutes, centrifuge, and separate the supernatant liq-uid. Combine all the extracts, add diluted methanol (7 in 10)to make exactly 50 mL, and use this solution as the samplesolution. Separately, weigh accurately about 10 mg of Sen-noside A Reference Standard, dissolve in a solution of sodi-um hydrogen carbonate (1 in 100) to make exactly 20 mL,and use this solution as standard stock solution (1). Weigh


accurately about 10 mg of Sennoside B Reference Standard,dissolve in a solution of sodium hydrogen carbonate (1 in100) to make exactly 20 mL, and use this solution as stan-dard stock solution (2). Pipet 5 mL of the standard stock so-lution (1) and 10 mL of the standard stock solution (2), addmethanol to make exactly 50 mL, and use this solution as thestandard solution. Perform the test with exactly 10 mL eachof the sample solution and the standard solution as directedunder the Liquid Chromatography according to the follow-ing conditions. Determine the peak areas, ATa and ASa, ofsennoside A, and the peak areas, ATb and ASb, of sennosideB in each solution, calculate the amounts of sennoside A andsennoside B by the following equations, and designate thetotal as the amount of total sennosides.


＝WSa×ATa

ASa×

14

Amount (mg) of sennoside B (C42H38O20)

＝WSb×ATb

ASb×

12

WSa: Amount (mg) of Sennoside A Reference Standard,calculated on the anhydrous basis

WSb: Amount (mg) of Sennoside B Reference Standard,calculated on the anhydrous basis

Operating conditions－Detector: An ultraviolet aborption photometer (wave-




Mobile phase: Dissolve 2.45 g of tetra-n-heptylammoni-um bromide in 1000 mL of a mixture of diluted 1 molWLacetic acid-sodium acetate buŠer solution, pH 5.0 (1 in 10)and acetonitrile (17:8).


System performance: When the procedure is run with 10mL of the standard solution under the above operating con-ditions, sennoside B and sennoside A are eluted in this orderwith the resolution between these peaks being not less than15, and the number of theoretical plates of the peak of sen-noside A being not less than 8000.


Powdered Senna Leafセンナ末


Powdered Senna Leaf is the powder of Senna Leaf.It contains not less than 1.0z of total sennosides

[sennoside A (C42H38O20: 862.75) and sennoside B(C42H38O20: 862.75)], calculated on the basis of driedmaterial.


Identiˆcation(2) To 2 g of Powdered Senna Leaf add 40 mL of a mix-

ture of tetrahydrofuran and water (7:3), shake for 30minutes, and centrifuge. Transfer the supernatant liquid to aseparator, add 13 g of sodium chloride, and shake for 30minutes. Separate the water layer with undissolved sodiumchloride, and adjust the pH to 1.5 with 1 molWL hydrochlor-ic acid TS. Transfer this solution to another separator, shakewith 30 mL of tetrahydrofuran for 10 minutes, separate thetetrahydrofuran layer, and use this solution as the samplesolution. Separately, dissolve 1 mg of Sennoside A Refer-ence Standard in 1 mL of a mixture of tetrahydrofuran andwater (7:3), and use this solution as the standard solution.Spot 10 mL each of these solutions on a plate of silica gel forthin-layer chromatography. Develop the plate with a mix-ture of ethyl acetate, 1-propanol, water and acetic acid (100)(40:40:30:1) to a distance of about 15 cm, and air-dry theplate. Examine under ultraviolet light (main wavelength: 365nm): one spot among the spots from the sample solution anda red ‰uorescent spot from the standard solution show thesame color tone and the same Rf value.


Assay Weigh accurately about 0.5 g of Powdered SennaLeaf in a glass-stoppered centrifuge tube, add 25 mL ofdiluted methanol (7 in 10), shake for 30 minutes, centrifuge,and separate the supernatant liquid. To the residue adddiluted methanol (7 in 10) twice with a 10-mL portoin, shakefor 10 minutes, centrifuge, and separate the supernatant liq-uid. Combine all the extracts, add diluted methanol (7 in 10)to make exactly 50 mL, and use this solution as the samplesolution. Separately, weigh accurately about 10 mg of Sen-noside A Reference Standard, dissolve in a solution of sodi-um hydrogen carbonate (1 in 100) to make exactly 20 mL,and use this solution as standard stock solution (1). Weighaccurately about 10 mg of Sennoside B Reference Standard,dissolve in a solution of sodium hydrogen carbonate (1 in100) to make exactly 20 mL, and use this solution as stan-dard stock solution (2). Pipet 5 mL of the standard stock so-lution (1) and 10 mL of the standard stock solution (2), addmethanol to make exactly 50 mL, and use this solution as thestandard solution. Perform the test with exactly 10 mL each


of the sample solution and the standard solution as directedunder the Liquid Chromatography according to the follow-ing conditions. Determine the peak areas, ATa and ASa, ofsennoside A, and the peak areas, ATb and ASb, of sennosideB in each solution, calculate the amounts of sennoside A andsennoside B by the following equations, and designate thetotal as the amount of total sennoside.


＝WSa×ATa

ASa×

14

Amount (mg) of sennoside B (C42H38O20)

＝WSb×ATb

ASb×

12

WSa: Amount (mg) of Sennoside A Reference Standard,calculated on the anhydrous basis

WSb: Amount (mg) of Sennoside B Reference Standard,calculated on the anhydrous basis





Mobile phase: Dissolve 2.45 g of tetra-n-heptylammoni-um bromide in 1000 mL of a mixture of diluted 1 molWLacetic acid-sodium acetate buŠer solution, pH 5.0 (1 in 10)and acetonitrile (17:8).


System performance: When the procedure is run with 10mL of the standard solution under the above operating con-ditions, sennoside B and sennoside A are eluted in this orderwith the resolution between these peaks being not less than15, and the number of theoretical plates of the peak of sen-noside A being not less than 8000.


Sinomenium Stemボウイ



Swertia Herbセンブリ


Identiˆcation To 2 g of pulverized Swertia Herb add 10mL of ethanol (95), shake for 5 minutes, ˆlter, and use theˆltrate as the sample solution. Separately, dissolve 2 mg ofSwertiamarin Refereance Standard in 1 mL of ethanol (95),and use this solution as the standard solution. Perform thetest with these solutions as directed under the Thin-layerChromatography. Spot 10 mL each of the sample solutionand the standard solution on a plate of silica gel with com-plex ‰uorescent indicator for thin-layer chromatography.Develop the plate with a mixture of ethyl acetate, 1-propanoland water (6:4:3) to a distance of about 10 cm, and air-drythe plate. Examine under ultraviolet light (broad spectrumwavelength): one spot among several spots from the samplesolution and a red spot from the standard solution show thesame color tone and the same Rf value.

Powdered Swertia Herbセンブリ末


Identiˆcation To 2 g of Powdered Swertia Herb add 10 mLof ethanol (95), shake for 5 minutes, ˆlter, and use theˆltrate as the sample solution. Separately, dissolve 2 mg ofSwertiamarin Reference Standard in 1 mL of ethanol (95),and use this solution as the standard solution. Perform thetest with these solutions as directed under the Thin-layerChromatography. Spot 10 mL each of the sample solutionand the standard solution on a plate of silica gel with com-plex ‰uorescent indicator for thin-layer chromatography.Develop the plate with a mixture of ethyl acetate, 1-propanoland water (6:4:3) to a distance of about 10 cm, and air-drythe plate. Examine under ultraviolet light (broad spectrumwavelength): one spot among several spots from the samplesolution and a red spot from the standard solution show thesame color tone and the same Rf value.


Swertia and Sodium BicarbonatePowderセンブリ重曹散

Change the Identiˆcation (1) and (2) to read:

Identiˆcation (1) To 10 g of Swertia and Sodium Bicar-bonate Powder add 10 mL of ethanol (95), shake for 15minutes, ˆlter, and use the ˆltrate as the sample solution.Separately, dissolve 1 mg of Swertiamarin Reference Stan-dard in 1 mL of ethanol (95), and use this solution as thestandard solution. Perform the test with these solutions asdirected under the Thin-layer Chromatography. Spot 30 mLeach of the sample solution and the standard solution on aplate of silica gel with ‰uorescent indicator for thin-layerchromatography. Proceed as directed in the Identiˆcationunder Powdered Swertia Herb.

(2) To 0.5 g of Swertia and Sodium Bicarbonate Powderadd 10 mL of water. After stirring, centrifuge the mixturewith 500 revolutions per minute. Smear, using a small glassrod, the slide glass with a small amount of the precipitate,add 1 drop of a mixture of water and glycerin (1:1), and puta cover glass on it so that the tissue section spreads evenlywithout overlapping each other, taking precaution againstinclusion of bubbles, and use this as the preparation formicroscopic examination. If the precipitate separates intotwo layers, proceed with the upper layer in the same manner,and use as the preparation for microscopic examination.Heat the preparation for microscopic examination in a shorttime: the preparation reveals the yellow-green to yellow-brown, approximately spherical pollen grains with granularpatterns under a microscope. The pollen grains are 25 – 34mm in diameter.

Toad Venomセンソ


Identiˆcation To 1 g of pulverized Toad Venom add 10mL of acetone, shake for 10 minutes, ˆlter, and use theˆltrate as the sample solution. Separately, dissolve 5 mg ofresibufogenin for thin-layer chromatography in 5 mL of ace-tone, and use this solution as the standard solution. Performthe test with these solutions as directed under the Thin-layerChromatography. Spot 10 mL each of the sample solutionand the standard solution on a plate of silica gel for thin-lay-er chromatography, develop the plate with a mixture of cy-clohexane and acetone (3:2) to a distance of about 10 cm,and air-dry the plate. Spray evenly dilute sulfuric acid on theplate, and heat at 1059C for 5 minutes: one of several spotsobtained from the sample solution has the same color toneand the same Rf value with the blue-green spot obtainedfrom the standard solution.

Add the following:

Uncaria ThornUncariae Uncis Cum Ramulus

チョウトウコウ

Uncaria Thorn is, usually, the prickle of Uncariarhynchophylla Miquel, Uncaria sinensis Haviland orUncaria macrophylla Wallich (Rubiaceae).

Uncaria Thorn contains not less than 0.03z of totalrhynchophylline (rhynchophylline and hirstine), calcu-lated on the dried basis.

Description Uncaria Thorn is uncinate prickle or shortstem with opposite or single prickle; the prickle, 1 to 4 cm inlength, curved and acuminate; externally red-brown to darkbrown or yellow-brown, some one with hairs, the transversesection oblong to elliptical, light brown; stem thin and pris-matic square to cylindrical, 2 to 5 mm in diameter, external-ly, red-brown to dark brown or yellow-brown; the transversesection, square to elliptical; the pith light brown, square toelliptical; hard in texture.

Odorless and practically tasteless.Under a microscope, a transverse section of the prickle

reveals vascular bundles in the cortex, unevenly distributedand arranged in a ring. Parenchyma cells in the secondarycortex containing sand crystals of calcium oxalate.

Identiˆcation To 1 g of pulverized Uncaria Thorn add 20mL of methanol, boil under a re‰ux condenser on a waterbath for 5 minutes, and ˆlter. Evaporate the ˆltrate to dry-ness, add 5 mL of dilute acetic acid to the residue, warm themixture on a water bath for 1 minute, and ˆlter after cool-ing. Spot 1 drop of the ˆltrate on a ˆlter paper, air-dry,spray DragendorŠ's TS for spraying on it, and allow tostand: a yellow-red color develops.




Component determination Weigh accurately about 0.2 gof medium powdered Uncaria Thorn, transfer into a glass-stoppered centrifuge tube, add 30 mL of a mixture ofmethanol and dilute acetic acid (7:3), shake for 30 minutes,centrifuge, and separate the supernatant liquid. To theresidue add a mixture of methanol and dilute acetic acid(7:3) twice with a 10-mL portion, proceed in the same man-ner, and combine all of the supernatant liquid. To the com-bined liquid add a mixture of methanol and dilute acetic acid(7:3) to make exactly 50 mL, and use this as the sample solu-tion. Separately, weigh accurately about 5 mg of rhyn-chophylline for component determination, previously driedin a desiccator (silica gel) for 24 hours, and dissolve in a mix-ture of methanol and dilute acetic acid (7:3) to make exactly100 mL. Pipet 1 mL of this solution, add a mixture ofmethanol and dilute acetic acid (7:3) to make exactly 10 mL,


and use this solution as the standard solution (1). Separately,dissolve 1 mg of hirsutine in 100 mL of a mixture ofmethanol and dilute acetic acid (7:3), and use this as thestandard solution (2). Perform the test with exactly 20 mLeach of the sample solution and the standard solution (1)and (2) as directed under the Liquid Chromatography ac-cording to the following conditions, and determine the peakareas, ATa and ATb, of rhynchophylline and hirsutine ob-tained from the sample solution, and the peak area, AS, ofrhynchophylline from the standard solution (1).

Amount (mg) of total rhynchophylline

＝WS×ATa＋ 1.405ATb

AS×

120

WS: Amount (mg) of rhynchophylline for component de-termination





Mobile phase: Dissolve 3.85 g of ammonium acetate inabout 200 mL of water, add 10 mL of acetic acid (100) andwater to make 1000 mL, and add 350 mL of acetonitrile.

Flow rate: Adjust the ‰ow rate so that the retention timeof rhynchophylline is about 17 minutes.System suitability—

System performance: Dissolve 5 mg of rhynchophyllinefor component determination in 100 mL of a mixture ofmethanol and dilute acetic acid (7:3). To 5 mL of this solu-tion add 1 mL of ammonia solution (28), and re‰ux for 10minutes or heat at about 509C for 2 hours. After cooling, to1 mL of the solution so obtained add a mixture of methanoland dilute acetic acid (7:3) to make 5 mL. When the proce-dure is run with 20 mL of this solution under the above oper-ating conditions, the peak of isorhynchophylline is appearsin addition to the peak of rhynchophylline, and the resolu-tion between these peaks is not less than 1.5.

System repeatability: When the test is repeated 6 timeswith 20 mL of the standard solution (1) under the aboveoperating conditions, the relative standard deviation of thepeak area of rhynchophylline is not more than 1.5z.

Change to read:

Vitamin A OilビタミンA油

Vitamin A Oil is synthetic vitamin A esters dilutedwith ˆxed oils. It contains not less than 30,000 Vita-min A Units per g. It may contain suitable antiox-idants.

It contains not less than 90.0z and not more than120.0z of the labeled amount of vitamin A.

Description Vitamin A Oil is a yellow to yellow-brown,clear or slightly turbid oil. It is odorless or has a faint, char-acteristic odor.

It is decomposed upon exposure to air or light.

Identiˆcation Dissolve Vitamin A Oil, Retinol AcetateReference Standard and Retinol Palmitate Reference Stan-dard, equivalent to 15,000 Units, in 5 mL of petroleumether, and use these solutions as the sample solution, thestandard solution (1) and the standard solution (2), respec-tively. Perform the test with these solutions as directed un-der the Thin-layer Chromatography. Spot 5 mL each of thesample solution and the standard solutions (1) and (2) on aplate of silica gel for thin-layer chromatography. Developwith a mixture of cyclohexane and diethyl ether (12:1) to adistance of about 10 cm, and air-dry the plate. Spray evenlyantimony (III) chloride TS: the principal spot obtained fromthe sample solution has the same color tone and the same Rfvalue with the blue spot obtained from the standard solution(1) or the standard solution (2).

Purity (1) Acid－Dissolve 1.2 g of Vitamin A Oil in 30mL of a mixture of neutralized ethanol and diethyl ether(1:1), boil gently for 10 minutes under a re‰ux condenser,cool, and add 5 drops of phenolphthalein TS and 0.60 mL of0.1 molWL sodium hydroxide VS: a red color develops.

(2) Rancidity—No unpleasant odor of rancid oil is per-ceptible by warming Vitamin A Oil.



Nitrogen atmosphere.


Change to read:

Vitamin A Oil CapsulesVitamin A Capsules

ビタミンA油カプセル

Vitamin A Oil Capsules contain not less than 90.0zand not more than 130.0z of the labeled Units ofVitamin A.

Method of preparation Prepare as directed under Cap-sules, using Vitamin A Oil.

Description The content of Vitamin A Oil Capsules con-forms to the requirements of the Description under VitaminA Oil.

Identiˆcation Proceed the test with the content of VitaminA Oil Capsules as directed in the Identiˆcation under Vita-min A Oil.

Assay Weigh accurately 20 Vitamin A Oil Capsules, andopen the capsules to take out the content. Wash the capsuleswell with a small amount of diethyl ether, allow the capsulesto stand at ordinal temperature to vaporize the diethyl ether,and weigh accurately. Perform the test with the content asdirected under the Vitamin A Assay, and calculate the unitsof vitamin A per capsule. Before applying Method 1-1, it isnecessary to know which the sample is, retinol acetate orretinol palmitate.


Storage—Light-resistant.

15671567Supplement I, JP XIV Infrared Reference Spectra

Add the following:Part I

15681568 Supplement I, JP XIVInfrared Reference Spectra

15871587Supplement I, JP XIV Ultraviolet-visible Reference Spectra

Delete the following Ultraviolet-visible Reference Spectra:

Part I

Homochlorcyclizine Hydrochloride 1

Homochlorcyclizine Hydrochloride 2

15881588 Supplement I, JP XIVUltraviolet-visible Reference Spectra

Change to read:Part I


Add the following:Part I

16111611

General Information

Change to read:

1. Aristolochic acidAristolochic acid, which occurs in plants of Aristolochia-

ceae, is suspected to cause renal damage. It is also reportedto be oncogenic (see References).

Aristolochic acid toxicity will not be a problem if crudedrugs of the origin and parts designated in the JP are used,but there may be diŠerences in crude drug nomenclature be-tween diŠerent countries, and it is known that crude drugpreparations not meeting the speciˆcations of the JP are cir-culating in some countries. Consequently, when crude drugsor their preparations are used, it is important that thematerials should not include any plant containingaristolochic acid.

In the Supplement I to JP14, the test for aristolochic acidI was added to the Purity under Asiasarum Root, which con-sists of the rhizome and root. Because the aerial part of theplant may contain aristolochic acid and may have beenimproperly contaminated in Asiasarum Root.

It is considered that Akebia Stem, Sinomenium Stem andSaussurea Root do not contain aristolochic acid, unlessplants of origin other than that designated in the JP areused. However, contamination of aristolochic acid might oc-cur, as mentioned above. In this case, the test described inthe Purity under Asiasarum Root is useful for checking thepresence of aristolochic acid.References:

Drug & Medical Device Safety Information (No.161)(July, 2000).

New England Journal of Medicine (June 8, 2000).Mutation Research 515, 63–72 (2002).

Change to read:

5. International HarmonizationImplemented in the JapanesePharmacopoeia Fourteenth

EditionItems for which harmonization has been agreed among

the European Pharmacopoeia, the United States Phar-macopeia and the Japanese Pharmacopoeia are implement-ed in the Japanese Pharmacopoeia Fourteenth Edition (JP14). They are shown in the tables below. The column headedHarmonized items shows the harmonized items written inthe Pharmacopoeial Harmonization Agreement Document,and the column headed JP 14 shows the items as they appearin JP 14. In the Remarks column, notes on any diŠerencesbetween JP 14 and the agreement are shown as occasion de-mands.

1612

Harmonized items JP 14 Remarks

Bacterial Endotoxin Test

Apparatus

Preparation of Standard EndotoxinStock solution

Preparation of Standard Endotoxinsolution

Preparation of sample solutions

Determination of Maximum Valid Di-lution

Gel-clot technique(1) Preparatory testing(2) Limit test(3) Assay

Photometric techniques(1) Turbidimetric technique(2) Chromogenic technique(3) Preparatory testing(4) Assay

Reagents, Test SolutionsAmebocyte lysateLysate TSWater for bacterial endotoxins test(BET)

Bacterial Endotoxins Test

Apparatus

Preparation of Standard EndotoxinStock Solution

Preparation of Standard EndotoxinSolution

Preparation of sample solutions

Determination of Maximum Valid Di-lution

Gel-clot techniques(1) Preparatory testing(2) Limit test(3) Assay

Photometric techniques(1) Turbidimetric technique(2) Chromogenic technique(3) Preparatory testing(4) Assay

Reagents, Test SolutionsLysate reagentLysate TSWater for bacterial endotoxins test

Note: The method for decision of the limit for bacterial endotoxins was agreed between the three pharmacopoeias, but in theDecision of Limit for Bacterial Endotoxins under the General Information in JP 14, the maximum adult dose is calculatedbased on an average body mass of an adult of 60 kg.

1612 Supplement I, JP XIVGeneral Information

1613


Residue on IgnitionWSulphated AshTest

(Introduction)

Residue on Ignition Test

(Introduction) Explanation of JP's particular expres-sions in the introduction: The descrip-tion, for example, ``not more than 0.10z (1 g),'' in a monograph, indicatesthat the mass of the residue is not morethan 1.0 mg per g of the substance to betested in which about 1 g of the sub-stance is weighed accurately and ignitedby the procedure described below, and`àfter drying'' indicates that the sam-ple is tested after being dried under theconditions speciêd in the test for Losson drying.

Procedure Procedure Explanation of JP's particular expres-sions for taking the sample: When thequantity of the sample to be taken is in-dicated in a volume, pipet exactly theamount directed in the monograph andtransfer to the above crucible. Whendirected as `àfter evaporating,'' heatproperly to evaporate the solution.


Sodium Chloride Sodium Chloride

Deˆnition limits of the content

Identiˆcation A Identiˆcation (1)

Identiˆcation B Identiˆcation (2)

Acidity or alkalinity Purity (2) Acidity or alkalinity

Bromides Purity (5) Bromides

Ferrocyanides Purity (7) Ferrocyanides

Iodides Purity (6) Iodides

Nitrites not speciêd

Phosphates Purity (4) Phosphates

Sulphates Purity (3) Sulfates

Aluminium not speciêd

Barium Purity (10) Barium

Iron Purity (9) Iron

Magnesium and alkaline-earth metals Purity (11) Magnesium and alkaline-earth materials

Potassium not speciêd

Loss on drying Loss on drying

Bacterial endotoxins not speciêd

Assay Assay

1613Supplement I, JP XIV General Information

1614


Citric Acid Monohydrate Citric Acid


Identiˆcation Identiˆcation

Appearance of solution Purity (1) Clarity and color of solu-tion

Readily carbonisable substances Purity (5) Readily carbonizable sub-stances

Oxalic acid Purity (3) Oxalate



Water Water

Sulphated ash Residue on ignition


Assay Assay

Storage Containers and storage


Citric Acid, Anhydrous Anhydrous Citric Acid


Identiˆcation Identiˆcation

Appearance of solution Purity (1) Clarity and color of solu-tion

Readily carbonisable substances Purity (5) Readily carbonizable sub-stances

Oxalic acid Purity (3) Oxalate



Water Water

Sulphated ash Residue on ignition


Assay Assay

Storage Containers and storage


1615


General Information

Sodium Dodecyl Sulphate Poly-acrylamide Gel Electrophoresis(SDS-PAGE)

General Information

SDS-Polyacrylamide Gel Electrophore-sis

Characteristics of PolyacrylamideGels

1. Characteristics of PolyacrylamideGels

Denaturing Polyacrylamide Gel Elec-trophoresis

Reducing conditions

Non-reducing conditions

2. Polyacrylamide Gel Electrophore-sis under Denaturing Conditions

1) Reducing conditions

2) Non-reducing conditions

Characteristics of DiscontinuousBuŠer System Gel Electrophoresis

3. Characteristics of DiscontinuousBuŠer System Gel Electrophoresis

Preparing Vertical DiscontinuousBuŠer SDS Polyacrylamide Gels

Assembling of the gel moulding cas-sette

Preparation of the gel

4. Preparing Vertical DiscontinuousBuŠer SDS-Polyacrylamide Gels

1) Assembling of the gel mouldingcassette

2) Preparation of the gel

Mounting the gel in the electrophore-sis apparatus and electrophoreticseparation

3) Mounting the gel in the electro-phoresis apparatus and electro-phoretic separation

Detection of Protein in Gels

Coomassie staining

Silver staining

5. Detection of Proteins in Gels

1) Coomassie staining

2) Silver staining

Drying of Stained SDS Poly-acrylamide Gels

6. Drying of Stained SDS-Polyacrylamide Gels

Molecular-Mass Determination 7. Molecular-Mass Determination

Validation of the Test 8. Suitability of the Test

Quantiˆcation of Impurities 9. Quantiˆcation of Impurities

Reagents, Test Solutions Test Solutions

Blocking solution Blocking TS

Coomassie staining solution Coomassie staining TS

Destaining solution Destaining TS

Developer solution Developer TS

Fixing solution Fixing TS

Silver nitrate reagent Silver nitrate TS for silver staining

Trichloroacetic acid reagent Trichloroacetic acid TS for ˆxing

Table 1－ Preparation of resolvinggel

Table 2－ Preparation of stacking gel

Table 1. Preparation of resolving gel

Table 2. Preparation of stacking gel



Change to read:

12. Preservatives-EŠectivenessTests

The purpose of the Preservatives-EŠectiveness Tests is toassess microbiologically the preservative e‹cacy, either dueto the action of product components themselves or anyadded preservative(s), for multi-dose containers. The e‹ca-cy of the preservatives is assessed by direct inoculation andmixing of the test strains in the product, and titration of sur-vival of the test strains with time.

Preservatives must not be used solely to comply withGMP for drugs or to reduce viable aerobic counts. In addi-tion, preservatives themselves are toxic substances. There-fore, preservatives must not be added to products inamounts which might jeopardize the safety of humanbeings, and consideration must be given to minimizing theamounts of preservative used. These tests are commonlyused to verify that products maintain their preservative eŠec-tiveness at the design phase of formulation or in the case ofperiodic monitoring. Although these tests are not performedfor lot release testing, the e‹cacy of the preservative presentin the product packaged in the ˆnal containers should beveriˆed throughout the entire dating period.

1. Products and their CategoriesThe products have been divided into two categories for

these tests. Category I products are those made with aqueousbases or vehicles, and Category II products are those madewith nonaqueous bases or vehicles. Oil-in-water emulsionsare considered Category I products, and water-in-oil emul-sions Category II products. Category I is further dividedinto three subtypes depending on the dosage forms.

Category IA: Injections and other sterile parenteralsCategory IB: Nonsterile parenteralsCategory IC: Oral products made with aqueous bases (in-

cluding syrup products to be dissolved or suspended beforeuse)

Category II: All the dosage forms listed under Category Imade with nonaqueous bases or vehicles.

2. Test Microorganisms and Culture MediaThe following strains or those considered to be equivalent

are used as the test microorganisms.Escherichia coli ATCC 8739, NBRC 3972Pseudomonas aeruginosa ATCC 9027, NBRC 13275Staphylococcus aureus ATCC 6538, NBRC 13276Candida albicans ATCC 10231, NBRC 1594, JCM 2085Aspergillus niger ATCC 16404, NBRC 9455These test microorganisms are representative of those that

might be found in the environment in which the product ismanufactured, used or stored, and they are also recognizedas opportunistic pathogens. In addition to these strainsdesignated as test microorganisms, it is further recommend-ed to use strains that might contaminate the product andgrow on or in it, depending on its characteristics. For the testmicroorganisms received from coordinated collections of

microorganisms, one passage is deˆned as the transfer ofmicroorganisms from an established culture to fresh medi-um, and microorganisms subjected to not more than ˆvepassages should be used for the tests. Single-strainchallenges rather than mixed cultures should be used. Thetest strains can be harvested by growth on solid agar or liq-uid media.

Cultures on agar plate media: Inoculate each of the ˆvetest strains on the surface of agar plates or agar slants. Forgrowth of bacteria, use Soybean-Casein Digest Agar Medi-um, and for yeasts and moulds, use Sabouraud Agar,Glucose-Peptone Agar or Potato Dextrose Agar Medium.Incubate bacterial cultures at 309C to 359C for 18 to 24hours, the culture of C. albicans at 209C to 259C for 40 to 48hours and the culture of A. niger at 209C to 259C for oneweek or until good sporulation is obtained. Harvest thesecultured cells aseptically using a platinum loop, etc. Suspendthe collected cells in sterile physiological saline or in 0.1zpeptone water and adjust the viable cell count to about 108

microorganisms per mL. In the case of A. niger, suspend thecultured cells in sterile physiological saline or 0.1z peptonewater containing 0.05 wWvz of polysorbate 80 and adjustthe spore count to about 108 per mL. Use these suspensionsas the inocula.

Liquid cultures: After culturing each of the four strainsexcept for A. niger in a suitable medium, remove the medi-um by centrifugation. Wash the cells in sterile physiologicalsaline or 0.1z peptone water and resuspend them in thesame solution with the viable cell or spore count of the in-oculum adjusted to about 108 per mL.

When strains other than the ˆve listed above are cultured,select a culture medium suitable for growth of the strain con-cerned. The cell suspension may also be prepared by amethod suitable for that strain. Use the inoculum suspen-sions within 24 hours after they have been prepared from thecultivations on agar plate media or in liquid media. Store theinoculum suspensions in a refrigerator if it is not possible toinoculate them into the test specimens within 2 hours. Ti-trate the viable cell count of the inocula immediately beforeuse, and then calculate the theoretical viable cell count permL (g) of the product present just after inoculation.

3. Test Procedure3.1 Category I products

Inject each of the cell suspensions aseptically into ˆve con-tainers containing the product and mix uniformly. When it isdi‹cult to inject the cell suspension into the container asep-tically or the volume of the product in each container is toosmall to be tested, transfer aseptically a su‹cient volume ofthe product into each of alternative sterile containers, andmix the inoculum. When the product is not sterile, incubateadditional containers containing the uninoculated productas controls and calculate their viable cell counts (the viablecounts of bacteria and those of yeasts and moulds). A sterilesyringe, spatula or glass rod may be used to mix the cell sus-pension uniformly in the product. The volume of the suspen-sion mixed in the product must not exceed 1W100 of thevolume of the product. Generally, the cell suspension is in-


oculated and mixed so that the concentration of viable cellsis 105 to 106 cells per mL or per gram of the product. Incu-bate these inoculated containers at 209C to 259C with pro-tection from light, and calculate the viable cell count of 1mL or 1 g of the product taken at 0, 14 and 28 days subse-quent to inoculation. Record any marked changes (e.g.,changes in color or the development of a bad odor) when ob-served in the mixed samples during this time. Such changesshould be considered when assessing the preservative e‹cacyof the product concerned. Express sequential changes in theviable counts as percentages, with the count at the start ofthe test taken as 100. Titration of the viable cell counts isbased, in principle, on the Pour Plate Methods in ``Microbi-al Limit Tests''. In this case, conˆrm whether any an-timicrobial substance is present in the test specimen. If aconˆrmed antimicrobial substance needs to be eliminated,incorporate an eŠective inactivator of the substance in thebuŠer solution or liquid medium to be used for dilution ofthe test specimen, as well as in the agar plate count medium.However, it is necessary to conˆrm that the inactivator hasno eŠect on the growth of the microorganisms. When the oc-currence of the preservative or the product itself aŠects titra-tion of the viable cell count and there is no suitable inactiva-tor available, calculate the viable cell counts by the Mem-brane Filtration Method in ``Microbial Limit Tests''.3.2 Category II products

The procedures are the same as those described for Cate-gory I products, but special procedures and considerationsare required for both uniform dispersion of the test microor-ganism in the product and titration of viable cell counts inthe samples.

For semisolid ointment bases, heat the sample to 459C to509C until it becomes oily, add the cell suspension and dis-perse the inoculum uniformly with a sterile glass rod or spat-ula. Surfactants may also be added to achieve uniform dis-persion, but it is necessary to conˆrm that the surfactantadded has no eŠect on survival or growth of the testmicroorganisms and that it does not potentiate the preserva-tive e‹cacy of the product. For titration of the viable cellcount, a surfactant or emulsiˆer may be added to dispersethe product uniformly in the buŠer solution or liquid medi-um. Sorbitan monooleate, polysorbate 80 or lecithin may beadded to improve miscibility between the liquid medium andsemisolid ointments or oils in which test microorganismswere inoculated. These agents serve to inactivate or neutral-ize many of the most commonly used preservatives.

4. InterpretationInterpret the preservative e‹cacy of the product accord-

ing to Table 1. When the results described in Table 1 are ob-tained, the product examined is considered to be eŠectivelypreserved. There is a strong possibility of massive microbialcontamination having occurred when microorganisms otherthan the inoculated ones are found in the sterile product tobe examined, and caution is required in the test proceduresandWor the control of the manufacturing process of theproduct. When the contamination level in a nonsterileproduct to be examined exceeds the microbial enumeration

limit speciˆed in ``Microbial Attributes of Nonsterile Phar-maceutical Products'' in General Information, caution isalso required in the test procedures andWor the control of themanufacturing process of the product.

Table 1. Interpretation criteria by product category

Productcategory Microorganisms

Interpretation criteria

After 14 days After 28 days

Category IA

Bacteria 0.1z of inocu-lum count orless

Same or lessthan level after14 days

YeastsWmoulds Same or lessthan inoculumcount

Same or lessthan inoculumcount

Category IB

Bacteria 1z of inoculumcount or less




Category IC

Bacteria 10z of inocu-lum count orless




Category II

Bacteria Same or lessthan inoculumcount




5. Culture MediaCulture media and buŠer solution used for Preservatives-

EŠectiveness Tests are described below. Other media may beused if they have similar nutritive ingredients and selectiveand growth-promoting properties for the microorganisms tobe tested.

Soybean Casein Digest Agar MediumCasein peptone 15.0 gSoybean peptone 5.0 gSodium chloride 5.0 gAgar 15.0 gWater 1000 mL

Mix all of the components and sterilize at 1219C for 15 –20 minutes in an autoclave. pH after sterilization: 7.1 – 7.3.

Sabouraud Glucose Agar MediumPeptone (animal tissue and casein) 10.0 gGlucose 40.0 gAgar 15.0 gWater 1000 mL

Mix all of the components and sterilize at 1219C for 15 –


20 minutes in an autoclave. pH after sterilization: 5.4 – 5.8.

Glucose Peptone (GP) Agar MediumGlucose 20.0 gYeast extract 2.0 gMagnesium sulfate heptahydrate 0.5 gPeptone 5.0 gMonobasic potassium phosphate 1.0 gAgar 15.0 gWater 1000 mL


Potato Dextrose Agar MediumPotato extract 4.0 gGlucose 20.0 gAgar 15.0 gWater 1000 mL


0.1z Peptone WaterPeptone 1.0 gSodium chloride 8.0 gWater 1000 mL


Add the following:

17. Basic Requirements for ViralSafety of BiotechnologicalWBiological Products listed in

Japanese PharmacopoeiaIntroduction

The primary role of speciˆcation of biotechnologicalWbio-logical products listed in Japanese Pharmacopoeia (JP) isnot only for securing quality control or consistency of thequality but also for assuring their e‹cacy and safety. In themeantime, the requirements to assure quality and safety ofdrugs have come to be quite strict recently, and a rigid atti-tude addressing safety assurance is expected for biotechno-logicalWbiological products. The key points for quality andsafety assurance of biotechnologicalWbiological products areselection and appropriate evaluation of source material, ap-propriate evaluation of manufacturing process and main-tenance of manufacturing consistency, and control ofspeciˆc physical properties of the products. Now, how to as-sure quality and safety of such drugs within a scope of JPhas come to be questioned. This General Informationdescribes what sorts of approaches are available to overcomethese issues.

It is desired that quality and safety assurance of JP listedproducts are achieved by state-of-the-art methods and con-cepts which re‰ect progress of science and accumulation ofexperiences. This General Information challenges to show

the highest level of current scientiˆc speculation. It is expect-ed that this information will contribute to promotion ofscientiˆc understanding of quality and safety assurance ofnot only JP listed products but also the other biotechnologi-calWbiological products and to promotion of active discus-sion of each O‹cial Monograph in JP.

1. Fundamental measures to ensure viral safety of JP listedbiotechnologicalWbiological products

The biotechnologicalWbiological product JP includes theproducts derived from living tissue and body ‰uid (urine,blood, etc.) of mammals, etc. In near future, protein drugsderived from cell lines of human or animal origin (e.g.,recombinant DNA drug, cell culture drug) will be included.The fundamental measures required for comprehensive viralsafety of JP listed biotechnologicalWbiological products areas follows: 1) acquaintance of possible virus contamination(source of contamination); 2) careful examination of eligibil-ity of raw materials and their sources, e.g. humanWanimal,and thorough analysis and screening of the sample chosen asa substrate for drug production (e.g., pooled body ‰uid, cellbank, etc.) to determine any virus contamination and deter-mination of type and nature of the virus, if contaminated; 3)evaluation to determine virus titer and virus-like particleshazardous to human, if exists; 4) selection of productionrelated material (e.g., reagent, immune antibody column)free from infectious or pathogenic virus; 5) performance ofvirus free test at an appropriate stage of manufacturing in-cluding the ˆnal product, if necessary; 6) adoption of eŠec-tive viral clearance method in the manufacturing process toremoveWinactivate virus. Combined method sometimesachieves higher level of clearance; 7) development of adeliberate viral clearance scheme; 8) performance of the testto evaluate viral removal and inactivation. It is consideredthat the stepwise and supplemental adoption of the saidmeasures will contribute to ensure viral safety and its im-provement.

2. Safety assurance measures described in the O‹cialMonograph and this General Information

As mentioned in above 1, this General Informationdescribes, in package, points to be concerned with and con-crete information on the measures taken for viral safety ofJP listed products. Except where any speciˆc caution is pro-vided in O‹cial Monograph of a product in question, O‹-cial Monograph provides in general that ``Any raw material,substrate for drug production and production relatedmaterial used for production of drug should be derived fromhealthy animals and should be shown to be free of latent vi-rus which is infectious or pathogenic to human'', ``Cell lineand culture method well evaluated in aspects of appropriate-ness and rationality on viral safety are used for production,and the presence of infectious or pathogenic latent virus tohuman in process related materials derived from living or-ganisms should be denied''. and ``biotechnologicalWbiologi-cal drug should be produced through a manufacturing proc-ess which is capable of removing infectious or pathogenic vi-rus'', etc., to raise awareness on viral safety and on necessityto conduct test and process evaluation for viral safety.


3. Items and contents described in this General Informa-tion

As for viral safety of protein drug derived from cell line ofhuman or animal origin, there is a Notice in Japan entitled``Viral safety evaluation of biotechnology products derivedfrom cell lines of human or animal origin'' (Iyakushin No.329 issued on February 22, 2000 by Director, Evaluation andLicensing Division, Pharmaceutical and Medical SafetyBureau, Ministry of Health and Welfare) to re‰ect the inter-nationally harmonized ICH Guideline, and as for bloodplasma protein fraction preparations, there is a documententitled ``Guideline for ensuring viral safety of blood plasmaprotein fraction preparations''. This General Informationfor ensuring viral safety of JP listed biotechnologicalWbio-logical products has been written, referencing the contentsof those guidelines, to cover general points and their detailsto be concerned for ensuring viral safety of not only JP list-ed biotechnologicalWbiological products but also allproducts which would be listed in JP in future, i.e., biologi-cal products derived from living tissue and body ‰uid, suchas urine, and protein drugs derived from cell line of humanor animal origin (Table 1).

Table 1. Items described in General Informationfor Viral Safety Assurance of JP listedBiotechnologicalWBiological Product

I. Introduction1. Fundamental measures to ensure viral safety of JP list-

ed biotechnologicalWbiological products2. Safety assurance measures described in the O‹cial

Monograph and this General Information3. Items and contents described in this General Informa-

tionII. General Matters

1. Purpose2. Background3. Unknown risk on the measures taken for ensuring viral

safety4. Applicable range5. Possible viral contamination to a JP listed biotechno-

logicalWbiological product (source of virus contamina-tion)

6. Basis for ensuring viral safety7. Limit of virus test8. Roles of viral clearance studies

III. Raw materialWsubstrate for drug production1. Issues relating to animal species and its region as a

source of raw materialWsubstrate for drug productionand countermeasures to be taken thereto

2. Qualiˆcation evaluation test on human or animal as asource of raw materialWsubstrate for drug production

IV. Points of concern with respect to manufacturing andvirus testing

1. Virus test conducted in advance of puriˆcation process2. Virus test as an acceptance test of an intermediate

material, etc.3. Virus test on a ˆnal product

V. Process evaluation on viral clearance

1. Rationale, objective and general items to be concernedwith respect to viral clearance process evaluation

2. Selection of virus3. Design of viral clearance studies4. Interpretation of viral clearance studies

1) Evaluation of viral clearance factor2) Calculation of viral clearance index3) Interpretation of results and items to be concerned at

evaluationVI. Statistics

1. Statistical considerations for assessing virus assays2. Reproducibility and conˆdence limit of viral clearance

studiesVII. Re-evaluation of viral clearanceVIII. Measurement for viral clearance studies

1. Measurement of virus infective titer2. Testing by nucleic-acid ampliˆcation test (NAT)

IX. Reporting and preservationX. Others

3.1 PurposeThe purpose of this document is to propose the compre-

hensive concepts of the measures to be taken for ensuringviral safety of biotechnologicalWbiological products derivedfrom living tissue or body ‰uid of mammals, etc. and of pro-tein drugs derived from cell lines of human or animal origin.That is to say, this document describes the measures and thepoints of concern on the items, such as consideration ofthe source of virus contamination; appropriate evaluationon eligibility at selecting the raw material and on qualiˆca-tion of its source, e.g. human or animal; virus test, and itsanalysis and evaluation at a stage of cell substrate for drugproduction; appropriate evaluation to choose productrelated materials derived from living organisms (e.g. rea-gent, immune antibody column, etc.); conduct of neces-sary virus test on the product at an appropriate stage ofmanufacturing; development of viral clearance testscheme; performance and evaluation of viral clearancetest. This document is also purposed to comprehensivelydescribe in details that supplemental and combining adop-tion of the said measures will contribute to secure viral safe-ty and its improvement.3.2 Background

One of the most important issues to be cautioned for safe-ty of a biological product, which is directly derived from hu-man or animal, or of a protein drug, which is derived fromcell line of human or animal origin (recombinant DNA der-ived product, cell culture derived product, etc.), is risk of vi-rus contamination. Virus contamination may cause serioussituation at clinical use once it occurs. Virus contaminationmay be from a raw material or from a cell substrate for drugproduction, or may be from an adventitious factor in-troduced to the manufacturing process.

JP listed biological drugs or protein drugs derived fromcell line have achieved drastic contribution to the medical so-ciety, and to date, there has not been any evidence of anysafety problem on them caused by virus. But, social require-ment of health hazard prevention is strong, and it is now


very important to prevent accidental incidence, takingsecurity measures carefully supported by scientiˆc rationali-ty. It is always great concern among the persons involvedthat under what sort of viewpoint and to what extent wehave to pursue for ensuring viral safety of a biotechnologicalWbiological product. Before discussing these issues, two fun-damental points have to be reconˆrmed. One is that; wehave to consider scientiˆc, medical, and social proˆles adrug has. In other words, ``Medicine is a social asset which isutilized in medical practice paying attention to the risk andbeneˆt from the standpoints of science and society''. It is thedestine and the mission of the medicalWpharmaceutical soci-ety to realize prompt and stable supply of such a social asset,drug, among the medical work front to bring gospel to thepatients.

The other is that; issue of viral safety is independent fromsafety of the components of a drug per se (narrow sense ofsafety). It is important to consider that this is the matter ofgeneral safety of drug (broad sense of safety). In case of adrug which has been used for a long time in the medicalfront, such as a JP listed product, its broad sense of safety isconsidered to have been established epidemiologically, andits usage past records have a great meaning. However, diŠer-ent from safety of drug per se (its components), taking intoaccount any possibility of virus contamination, we have tosay that only the results accumulated can not always assureviral safety of a drug used in future. Accordingly, the basisfor securing broad sense of viral safety of JP listedbiotechnologicalWbiological products is to pay every atten-tion to the measures to take for prevention, while evaluatingthe accumulated results.

Adopting strict regulations and conducting tests at maxi-mum level to the extent theoretically considered may be theways oŠ assuring safety, but applying such way generally,without su‹cient scientiˆc review of the ways and evalua-tion of usage results, causes excessive requirement of regula-tion and test not having scientiˆc rationality. As the results,eŠective and prompt supply of an important drug, alreadyhaving enough accumulation of experiences, to the medicalwork front will be hampered, and the drug, a social asset,may not to be utilized eŠectively. Medicine is a sword used inmedical êld having double-edge named eŠectiveness andsafety. EŠectiveness and safety factors have to be derived asthe fruits of leading edge of science, and relatively evaluatedon a balance sheet of usefulness. Usefulness evaluationshould not be unbalanced in a way that too much emphasisis placed on safety concern without back-up of appropriatescientiˆc rationality. A drug can play an important role as asocial asset only when well balanced appropriate scientiˆcusefulness evaluation in addition to social concern of the ageare given. In other words, drug is a common asset utilized bysociety for medication as a fruit of science of the age, andthe key point of its utilization lies on a balance of risk andbeneˆt produced from scientiˆc and social evaluation. So,those factors have to be taken into account when target andpursuance levels for ensuring viral safety of a JP listedbiotechnologicalWbiological product are reviewed.

And, in general, the risk and beneˆt of drugs should be

considered with the relative comparison to alternative drugsor medical treatment. The usefulness of certain drug shouldbe reviewed ˆnally after the competitive assessment on therisk and beneˆt on the alternative drugs, relevant drugs andWor alternative medical treatment.

Under such background, the purpose of this article is todescribe the scientiˆc and rational measures to be taken forensuring viral safety of JP listed biotechnologicalWbiologicalproducts. Giving scientiˆc and rational measures mean that;appropriate and eŠective measures, elaborated from the cur-rent scientiˆc level, are given to the issues assumable underthe current scientiˆc knowledge. In other words, possiblecontaminant virus is assumed to have the natures of genus,morph, particle size, physicalWchemical properties, etc.which are within the range of knowledge of existing virolo-gy, and is those assumed to exist in human and animal, tis-sue and body ‰uid, which are the source of biotechnologicalWbiological product, reagent, material, additives, etc. Ac-cordingly, viral clearance studies using a detection methodwhich target those viruses have to be designed.3.3 Unknown risk on the measures taken for ensuring viralsafety

There are known and unknown risks.It is easy to determine a test method and an evaluation

standard on the known risk, which exists in the drug per se(pharmaceutical component) or inevitably exists due to aquality threshold, and quantiˆcation of such risk is possible.In other words, it is easy to evaluate the known risk on abalance sheet in relation to the beneˆt, and we can say thatvaluation even in this respect has been established to someextent.

On the other hand, as for the unknown risk which is in-evitable for ensuring viral safety, the subject of the risk cannot be deˆned and quantitative concept is hard to introduce,and, therefore, taking a counter measure and evaluating itseŠect are not so easy. Therefore, this is the subject to bechallenged calling upon wisdom of the related parties amongthe society of drug.

Talking about the unknown risk, there are view pointsthat say `Ìt is risky because it is unknown.'' and ``What arethe unknowns, and how do we cope with them in ensuringsafety?''.

The view of `Ìt is risky because it is unknown.'' is alreadynothing but a sort of evaluation result, and directly connectsto a ˆnal decision if it can be used as a drug. Such evaluationWdecision has to be made based upon a rational, scientiˆc orsocial judgment.

For example, in the case that `Ìn a manufacturing processof drug, virus, virus-like particle or retrovirus was detected,but its identiˆcation could not be conˆrmed, and, therefore,its risk can not be denied.'', the evaluation of `Ìt is risky be-cause it is unknown.'' is scientiˆcally rational and reasona-ble. On the other hand, however, if we reach a decision of`Ìt is risky because it is unknown.'' due to the reason that`Ìn a manufacturing process of drug, virus, virus-like parti-cle or retrovirus was not detected, but there is a `concern'that something unknown may exist.'', it can not be said thatsuch evaluation is based upon a rational, scientiˆc or social


judgment. It goes without saying that the utmost care has tobe taken for viral safety, but the substance of `concern' hasto be at least clearly explainable. Otherwise, the `concern'may result in causing contradiction in the meaningful mis-sion to utilize a social asset, drug, in medical practice.

From scientiˆc view point, we should not be narrow mind-ed by saying `ìt is risky'' because ``there is a `concern' thatsomething unknown may exists'', but challenge to clarify thesubject of ``What is unknown, and how to cope with it forensuring safety'' using wisdom. What is important at thetime is to deˆne ``what is unknown'' based upon currentscientiˆc knowledge. Only through this way, is it possible forus to elaborate the measures for ensuring safety.

Once we chase up the substance of unknown risk for viralsafety without premises of ``what is unknown'',`ùnknown'' will be an endless question because it theoreti-cally remains unresolved forever. If this kind of approach istaken, the issue and the measure can not be scientiˆcallyconnected to each other, which will result in the excessive re-quirement of regulation and of test to be conducted. Yet, itis unlikely that the measure which has no relation withscience will be eŠective to the subject of ``What is unknownis unknown.''

For example, ``what is unknown'' at the `èvaluation of apuriˆcation process which can completely clear up every vi-rus that contaminated in a manufacturing process'' shouldbe the subject of ``what sort of existing virus that contami-nated is unknown'', not on the subject of ``what sort ofvirus that exist in the world is unknown. In the former sub-ject, the premise of the study is based on all the knowledgeon viruses including DNAWRNA-virus, virus withWwithoutenvelope, particle size, physicalWchemical properties, etc.The premise is that the virus contaminated should be withinrange of existing wisdom and knowledge of virus such asspecies, type, nature, etc., even though the virus that con-taminated is unknown. Under such premise, when evalua-tion is made on a puriˆcation process to decide its capabilityof clearing a derived virus, which is within the range of exist-ing wisdom and learning, speciˆc viral clearance studiesdesigned to combine a few model viruses with diŠerent na-tures, such as type of nucleic acid, withWwithout envelope,particle size, physicalWchemical properties, etc., would beenough to simulate every sort of the virus already known,and will be a ``good measure for ensuring safety''.

The issue of ``the sort of viruses that exist in the world isunknown'' may be a future study item, but it is not an ap-propriate subject for the viral clearance test. Further, even ifthe subject of `ùnknown viruses, which have a particle sizesmaller than that of currently known viruses, may exists'' or`ùnknown viruses, which have special physicalWchemicalproperties that can not be matched to any of the currentlyknown viruses, may exists'' is set up as an armchair theory,any experimental work can not be pursued under the currentscientiˆc level, since such virus model is not available. Fur-ther, any viral clearance test performed by using the current-ly available methods and technologies will be meaningless``for ensuring safety'', since particle size or natures of suchspeculated virus are unknown. Likewise, any counter meas-

ures can not be taken on the subject of `ùnknown virus,which can not be detected by currently available screeningmethod, may exist'', and conducting any virus detection testat any stage will be useless ``for ensuring safety''.

The requirement of regulations or tests excessively overscientiˆc rationality will raise human, economical and tem-poral burden to the pharmaceutical companies, and will ad-versely aŠect prompt, eŠective and economical supply of adrug to the medical front. As drug is a sort of social asset,which has to be scientiˆcally evaluated, how to assure max-imization of its safety by means of scientiˆcally rational ap-proach at minimum human, economical and time resourcesis important.

It is also important to reconˆrm that achievement of thoseissues is on the premise that appropriate measures are takenon the supply source of drugs. For example, in a case of `Ìna manufacturing process of drug, virus, virus-like particle orretrovirus was not detected, but there is a `concern' thatsomething unknown may exist.'', appropriateness of thetest, which resulted in the judgment that ``virus, virus-likeparticle or retrovirus was not detected in a process of drugproduction'', should be a prerequisite premise when judgedby science standard ate the time. If there is any question onthe premise, it is quite natural that the question of ``there is a`concern' that unknown something may exist.'' will be eŠec-tive.3.4 Applicable range

This General Information is on JP listed biologicalproducts, derived from living tissue or body ‰uid, and pro-tein drugs, derived from human or animal cell line, that inJapan. In the case of protein drugs derived from human oranimal cell line, the products developed and approved be-fore enforcement of the Notice Iyakushin No. 329 entitled``Viral safety evaluation of biotechnology products derivedfrom cell lines of human or animal origin'' should have beentreated under the Notice had there been one, and it is inevita-ble that some products approved after the Notice might nothave been su‹ciently treated. It is expected that suchbiodrug will be su‹ciently examined to meet such GeneralInformation before being listed in JP. On the other hand,blood preparations listed in the biological products standardand covered by ``Guideline for securing safety of blood plas-ma protein fraction preparations against virus'', are out ofthe scope of this General Information. Further, in case of arelatively lower molecular biogenous substance, such as ami-no acid, saccharide and glycerin, and of gelatin, which iseven classiêd as infectious or pathogenic polymer, there arecases that viral contamination can not be considered due toits manufacturing or puriˆcation process, and that potentviral inactivationWremoval procedure that can not be appliedto protein, can be used, and, therefore, it is consideredreasonable to omit such substances from the subject for ap-plication. However, some part of this General Informationmay be used as reference. Further, a comprehensive assur-ance measure for viral safety is recommendable on abiotechnologicalWbiological product not listed in JP usingthis document as a reference so long as it is similar to thebiotechnologicalWbiological product JP.


3.5 Possible viral contamination to a JP listed biotechno-logicalWbiological product (source of virus contamination)

Promoting awareness of virus contamination to a JP list-ed biotechnologicalWbiological product (source of virus con-tamination) and citing countermeasure thereof are im-portant for eradicating any possible virus contamination andraising probability of safety assurance. Many biotechnologi-calWbiological products are produced from a ``substrate''which is derived from human or animal tissue, body ‰uid,etc. as an originWraw material, and in puriˆcation or phar-maceutical processing of such products column materials oradditives, which are living organism origin, are occasionallyused. Accordingly, enough safety measures should be takenagainst diŠusion of the contaminant virus. Further, as men-tioned in Notice Iyakushin No. 329, any protein drug der-ived from cell lines of human or animal origin should becarefully examined with respect to the risk of virus contami-nation through the cell line, the cell substrate for drugproduction, and through the manufacturing process appliedthereafter.

``Substrate for drug production'' is deˆned as a startingmaterial which is at a stage where it is deemed to be in a posi-tion to ensure qualityWsafety of an active substance. The``substrate for drug production'' is sometimes tissue, body‰uid, etc. of human or animal per se and pooled materialsuch as urine, and sometimes a material after some treat-ment. In many cases, it is considered rational that startingpoint of full-scale test, evaluation and control should be atthe stage of ``substrate for drug production''. The morestrict levels of test, evaluation and control achieved at thestage of ``substrate for drug production'' can more rational-ize evaluation and control of the raw material or individuallevel of upper stream. On the contrary, strict evaluation andcontrol of the raw material or individual level at an upperstream stage can rationalize tests, evaluation or quality con-trol at the stage of ``substrate for drug production''.

The measures taken for ensuring viral safety on abiotechnologicalWbiological product currently listed in JPcan be assumed from the provisions of manufacturingmethod, speciˆcation and test methods of each preparation.However, unitary principles or information with respect tothe measures to be taken for ensuring viral safety, totallyreviewing the entire process up to the ˆnal product rationallyand comprehensively, including sourceWraw materialWsub-strate, puriˆcation process, etc. have not been clariˆed. Themost important thing for ensuring viral safety is to takethorough measures to eliminate the risk of virus contamina-tion at any stage of source animal, raw material and sub-strate. Although not the cases of a biotechnologicalWbiologi-cal product, known examples of a virus contamination froma raw materialWsubstrate for drug production in old timesare Hepatitis A Virus (HAV) or Hepatitis C Virus (HCV)contamination in blood protein fraction preparations. It isalso well known that Human Immunodeˆciency Virus (HIV)infection caused by blood plasma protein fraction prepara-tions occurred in 1980s. The aim of this General Informa-tion is to show concrete guidelines for comprehensive viralsafety assurance of the JP listed biotechnologicalWbiological

products. The pathogenic infectious viruses, currentlyknown to contaminate to raw materials, etc. of drug andhave to be cautioned, are HIV, HAV, Hepatitis B Virus(HBV), HCV, Human T-Lymphotropic Virus (HTLV-IWII),Human Parvovirus B19, Cytomegalovirus (CMV), etc.BiotechnologicalWbiological products produced from rawmaterialWcell substrate derived from tissue or body ‰uid ofhuman or animal origin always have a risk of contaminationof pathogenic or other latent virus. Therefore, safety meas-ures should be thoroughly taken. There is also the case that amaterial, other than the biological component such as rawmaterialWsubstrate, causes virus contamination. Using en-zymatic or monoclonal antibody column or using albuminetc. as a stabilizer, is the example of the case, in which cau-tion has to be taken on risk of virus contamination from thesource animal or cell. Further, there is a possibility of con-tamination from environment or personnel in charge ofproduction or at handling of the product. So, caution has tobe taken on these respects as well.

In case of protein drugs derived from cell line of human oranimal origin, there may be cases where latent or persistentinfectious viruses (e.g., herpesvirus) or endogenousretroviruses exist in the cell. Further, adventitious virusesmay be introduced through the routes such as: 1) derivationof a cell line from an infected animal; 2) use of a virus todrive a cell line; 3) use of a contaminated biological reagent(e.g., animal serum components); 4) contamination duringcell handling. In the manufacturing process of drug, anadventitious virus may contaminate to the ˆnal productthrough the routes, such as 1) contamination through a rea-gent of living being origin, such as serum component, whichis used for culturing, etc.; 2) use of a virus for introductionof a speciˆc gene expression to code an objective protein; 3)contamination through a reagent used for puriˆcation suchas monoclonal antibody a‹nity column; 4) contaminationthrough an additive used for formulation production; 5)contamination at handling of cell and culture media, etc. Itis reported that monitoring of cell culture parameter may behelpful for early detection of an adventitious viral contami-nation.3.6 Basis for ensuring viral safety

Viral safety of a biotechnologicalWbiological product pro-duced from a raw materialWsubstrate, which derived fromtissue, body ‰uid, cell line, etc. of human or animal origin,can be achieved by supplemental and appropriate adoptionof the following plural methods.(1) Acquaintance of possible virus contamination (source

of contamination).(2) Careful examination of eligibility of the raw material

and its source, i.e., human or animal, thorough analy-sis and screening of the sample chosen as the substratefor drug production to determine virus contaminationand through examination of the type of virus and itsnature, if contaminated.

(3) Evaluation to determine hazardous properties of the vi-rus or virus-like particle to human, if exists.

(4) Choosing a product related material of living organismorigin (e.g., reagent, immune anti-body column, etc.)


which is free from infectious or pathogenic virus.(5) Conduct virus free test at an appropriate stage of

manufacturing including the ˆnal product, if necessary.(6) Adoption of an eŠective method to removeWinactivate

the virus in the manufacturing process for viral clear-ance. Combined processes sometimes achieve higherlevel of viral clearance.

(7) Develop a deliberate viral clearance scheme.(8) Conduct test and evaluation to conˆrm removalWinacti-

vation of the virus.Manufacturer is responsible for explaining rationality of

the way of approach adopted among the comprehensivestrategy for viral safety on each product and its manufactur-ing process. At the time, the approach described in thisGeneral Information shall be applicable as far as possible.3.7 Limit of virus test

Virus test has to be conducted to deˆne existence of virus,but it should be noted that virus test alone can not reach aconclusion of inexistence of virus nor su‹cient to securesafety of the product. Examples of a virus not being detectedare as follows: 1) Due to statistical reason, there is an inher-ent quantitative limit, such as detection sensitivity at lowerconcentration depends upon the sample size. 2) Generally,every virus test has a detection limit, and any negative resultof a virus test can not completely deny existence of a virus.3) A virus test applied is not always appropriate in terms ofspeciˆcity or sensitivity for detection of a virus which existsin the tissue or body ‰uid of human or animal origin.

Virus testing method is improved as science and technolo-gy progress, and it is important to apply scientiˆcally themost advanced technology at the time of testing so that itcan be possible to raise the assurance level of virus detection.It should be noted, however, that the limit as mentionedabove can not always be completely overcome. Further, riskof virus contamination in a manufacturing process can notbe completely denied, and, therefore, it is necessary toelaborate the countermeasure taken these eŠects into ac-count.

Reliable assurance of viral free ˆnal product can not beobtained only by negative test results on the raw materialWsubstrate for drug production or on the product in general, itis also necessary to demonstrate inactivationWremovalcapability of the puriˆcation process.3.8 Roles of viral clearance studies

Under the premises as mentioned in the preceding clausethat there is a limit of a virus test, that there is a possibilityof existence of latent virus in a raw materialWsubstrate fordrug production and that there is a risk of entry of a non-en-dogenous virus in a manufacturing process, one of the im-portant measures for viral safety is how to remove or inacti-vate the virus, which exists in a raw material, etc. and cannot be detected, or the virus, which is contingently contami-nated in a manufacturing process. The purpose of viralclearance study is to experimentally evaluate the viralremovalWinactivation capability of a step that mounted in amanufacturing process. So, it is necessary to conduct an ex-perimental scale spike test using an appropriate virus that isselected by taking account the properties, such as particle

size, shape, with or without envelope, type of nucleic acid(DNA type, RNA type), heat and chemical treatment toler-ance, etc., with an aim to determine removalWinactivationcapability of the virus that can not be detected in a rawmaterial or contingently contaminated.

As mentioned above, the role of viral clearance study is tospeculate removalWinactivation capability of a processthrough a model test, and it contributes to give scientiˆc ba-sis to assure that a biotechnologicalWbiological product ofhuman or animal origin has reached an acceptable level inaspect of viral safety.

At a viral clearance study, it is necessary to adopt an ap-propriate approach method which is deˆnitive and rationaland can assure viral safety of a ˆnal product, taking intoconsideration the source and the properties of the rawmaterialWsubstrate as well as the manufacturing process.

4. Raw materialWsubstrate for drug production4.1 Issues relating to animal species and its region as asource of raw materialWsubstrate for drug production andcountermeasures to be taken thereto

For manufacturing JP listed biotechnologicalWbiologicalproducts, which require measures for viral safety, a rawmaterialWsubstrate derived mainly from human, bovine,swine or equine is used, and it is obvious that such humanand animal has to be healthy nature. A wild animal shouldbe avoided, and it is recommended to use animals derivedfrom a colony controlled by an appropriate SPF (SpeciˆcPathogen-Free) condition and bred under a well deignedhygienic control, including appropriate control for preven-tion of microbial contamination and contaminationmonitoring system. If a meat standard for food is available,an animal meeting this standard has to be used. The type ofvirus to be concerned about depend on animal species, but itmay be possible to narrow down the virus for investigationby means of examining the hygiene control, applicability ofa meat standard for food, etc. On the other hand, even withthe animals of the same species, a diŠerent approach may benecessary depending upon the region where the specimen fora raw materialWsubstrate is taken. For example, in case ofobtaining raw materialWsubstrate from blood or otherspeciˆc region, it is necessary to be aware of the risk level, vi-rus multiplication risk, etc. which may speciˆcally exists de-pending upon its region. Such approach may be diŠerentfrom those applied to body waste such as urine, milk, etc. asa source of raw materialWsubstrate. Further, caution has tobe taken on transmissible spongiform encephalopathy (TSE)when pituitary gland, etc. is used as a raw material. Thisreport does not include detailed explanation on TSE, butrecommendations are to use raw material derived from 1)animals originated in the countries (area) where incidence ofTSE has not been reported; 2) animals not infected by TSE;or 3) species of animal which has not been reported on TSE.It is recommended to discuss the matters concerned withTSE with the regulatory authority if there is any unclearpoint.

Followings are the raw materialWsubstrate used formanufacturing biotechnologicalWbiological products in


Japan.(1) Biological products derived from human

Blood plasma, placenta, urine, etc. derived from humanare used as the sources of raw material of biotechnologicalWbiological product. As for these raw materials, there are 2cases: 1) Appropriateness can be conˆrmed by interview orby examination of the individual who supplies each rawmaterial, and 2) Such su‹cient interview or examination ofthe individual can not be made due to type of raw material.In case that su‹cient examination of individual level is notpossible, it is necessary to perform test to deny virus con-tamination at an appropriate manufacturing stage, for ex-ample, the stage to decide it as a substrate for drug produc-tion.(2) Biological products derived from animal besides hu-man

Insulin, gonadotropin, etc. are manufactured from bloodplasma or from various organs of bovine, swine and equine.(3) Protein drug derived from cell line of human or animalorigin

It is expected that protein drugs derived from cell line ofhuman or animal origin will be listed in JP in a near future.In the case of these products, a cell line of human or animalis the raw material per se, and the substrate for drug produc-tion is a cell bank prepared from cloned cell line (master cellbank or working cell bank). Examination at cell bank level isconsidered enough for viral safety qualiˆcation, but it goeswithout saying that the more appropriate and rationalqualiˆcation evaluation test of cell bank can be realizedwhen more information is available on the virus of thesource animal or on prehistory of driving the cell line, thebase of cell bank.4.2 Qualiˆcation evaluation test on human or animal as asource of raw materialWsubstrate for drug production(1) Biological products derived from human

Body ‰uid etc. obtained from healthy human must beused for biological products production. Further, in casethat interview or examination of the individual, who sup-plies the raw material, can be possible and is necessary, in-terview under an appropriate protocol and a serologic testwell evaluated in aspects of speciˆcity, sensitivity and ac-curacy have to be performed, so that only the raw material,which is denied latent HBV, HCV and HIV, will be used. Inaddition to the above, it is necessary to test for gene of HBV,HCV and HIV by a nucleic ampliˆcation test (NAT) wellevaluated in aspects of speciˆcity, sensitivity and accuracy.

In case of the raw material (e.g., urine), which can not betested over the general medical examination of the individualwho supplies the material, or of the raw material which is ir-rational to conduct individual test, the pooled raw material,as the substrate for drug production, has to be conducted atleast to deny existence of HBV, HCV and HIV, using amethod well evaluated in aspects of speciˆcity, sensitivityand accuracy, such as the antigen test or NAT.(2) Biological products derived from animal besides hu-man

The animal used for manufacturing biological product hasto be under appropriate health control, and has to be con-

ˆrmed of its health by various tests. Further, it is necessarythat the population, to which the animal belongs, has beenunder an appropriate breeding condition, and that no abnor-mal individual has been observed in the population. Further,it is necessary to demonstrate information or scientiˆc basiswhich can deny known causes infection or disease to human,or to deny such animal inherent latent virus by serologic testor by nucleic ampliˆcation test (NAT). The infectious virusthat is known to be common between human and animal,and known to cause infection in each animal are tentativelylisted in Table 2. It is necessary that the table is completedunder careful examination, and denial of all of them, bymeans of tests on individual animal, tissue, body ‰uid, etc.as a raw material, or on pooled raw material (as a direct sub-strate for drug production), is not always necessary. Table 2can be used as reference information, in addition to theother information, such as; source of animal, health condi-tion, health and breeding control, conformity to the meatstandard for food, etc., to elaborate to which virus whatkind of test has to be performed, and for which virus it is notalways necessary to test for, etc. It is important to clarifyand record the basis of choosing the virus and the test con-ducted thereof.

Table 2. Infectious viruses known to be commonbetween human and animal and known to

cause infection to each animal

bovine swine sheep goat equine

Cowpox virus w

Paravaccinia virus w w w w

Murry valley encephalitisvirus w w

Louping-ill virus w w w w

Wesselsbron virus w

Foot-and-mouth diseasevirus w w

Japanese encephalitisvirus w

Vesicular stomatitis virus w

Bovine papular stomatitisvirus w

Orf virus w

Borna disease virus w w

Rabies virus w w w w w

In‰uenza virus w

Porcine hepatitis E virus w

Encephalomyocarditisvirus w w

Rota virus w


Eastern equineencephalitis virus w

Western equineencephalitis virus w

Venezuelan equineencephalitis virus w

Morbillivirus w

Hendravirus w

Nipahvirus w

Transmissible gastroente-ritis virus w

Porcine respiratorycoronavirus w

Porcine epidemicdiarrheavirus w

Hemagglutinatingencephalomyelitis virus w

Porcine respiratory andreproductive complex virus w

Hog cholera virus w

Parain‰uenza 3 virus w

TelfanWTeschen diseasevirus w

Reoviruses w

Endogenous retrovirus w

Porcine adenovirus w

Porcine circovirus w

Porcine parvovirus w

Swine poxvirus w

Porcine cytomegalovirus w

Pseudorabies virus w

Russian spring summerencephalitis virus w w

Rift Valley fever virus w w

Crimean-Congo hemor-rhagic fever virus(Nairovirus)

w w w

Torovirus w

(3) Protein drug derived from cell line of human or animalorigin

It is important to conduct thorough investigation on latentendogenous and non-endogenous virus contamination in amaster cell bank (MCB), which is the cell substrate for drugproduction, in accordance with the Notice Iyakushin No.329 entitled ``Viral safety evaluation of biotechnologyproducts derived from cell lines of human or animal origin''.

Further, it is necessary to conduct an appropriate adventiti-ous virus test (e.g., in vitro and in vivo test) and a latent en-dogenous virus test on the cell at the limit of in vitro cell age(CAL) for drug production. Each WCB as a starting cellsubstrate for drug production should be tested for adventiti-ous virus either by direct testing or by analysis of cells at theCAL, initiated from the WCB. When appropriate non-en-dogenous virus tests have been performed on the MCB andcells cultured up to or beyond the CAL have been derivedfrom the WCB and used for testing for the presence ofadventitious viruses, similar tests need not be performed onthe initial WCB.

5. Points of concern with respect to manufacturing andvirus testing

To ensure viral safety of a biological product derived fromtissue, body ‰uid etc. of human or animal origin, it is neces-sary to exclude any possibility of virus contamination from araw material, such as tissue and body ‰uid, or a substrate,paying attention to the source of virus contamination asmentioned in above 3.5, and to adopt appropriate manufac-turing conditions and technologies in addition to enhance-ment of manufacturing environment, so that virus contami-nation in the course of process and handling and from oper-ators, facilities and environment can be minimized.

In addition to the above, eŠective virus test and viral inac-tivationWremoval technology, which are re‰ected by rapidprogress of science, have to be introduced. Adoption of twoor more steps with diŠerent principles is recommended forvirus inactivationWremoval process. Further, it is importantto minimize any possible virus derivation by using a reagent,which quality is equivalent to that of a drug. Examples of vi-rus inactivationWremoval processes are heating (It isreported that almost viruses are inactivated by heating at 55– 609C for 30 minutes with exceptions of hepatitis virus, etc.and that dry heating at 609C for 10 – 24 hours is eŠective incase of the products of blood or urine origin.), treatmentwith organic solventWsurfactant (SWD treatment), mem-brane ˆltration (15 – 50 nm), acid treatment, irradia-tion (g-irradiation, etc.), treatment with column chro-matograph (e.g. a‹nity chromatography, ion-exchangechromatography), fractionation (e.g. organic solvent orammonium sulfate fractionation), extraction.5.1 Virus test conducted in advance of puriˆcation process(1) Biological products derived from human

In many cases, samples for virus test before puriˆcationprocess are body ‰uid or tissue of individual collected as araw material, or its pooled material or extraction as a sub-strate. As mentioned in 4.2 (1), it is necessary to deny latentHBV, HCV and HIV by the test evaluated enough in aspectsof speciˆcity, sensitivity and accuracy. Even in a case that anon-puriˆed bulk before puriˆcation process is producedfrom a substrate, it is not always necessary to conduct virustest again at the stage before puriˆcation, so long as thepresence of any latent virus can be denied at the stage of sub-strate by an appropriate virus test, with cases where the non-puriˆed bulk is made from the substrate by adding any rea-gent etc. of living organisms origin are an exception.


(2) Biological products derived from animal besideshuman

Similar to (1) above, samples for virus test before puriˆca-tion process are, in many cases, body ‰uid or tissue of in-dividual collected as a raw material, or its pooled material orextraction as a substrate. In these cases, it is necessary tohave a data, which can deny latent virus of probable cause ofhuman infection or disease as mentioned in the above 4.2(2), or to have a result of serologic test or nucleic ampliˆca-tion test (NAT) evaluated enough in aspects of speciˆcity,sensitivity and accuracy. The concept, which is applied to acase that non-puriˆed bulk before puriˆcation process isproduced from substrate, is the same as those provided inthe above 4.2 (1).(3) Protein drug derived from cell line of human or animalorigin

Generally, substrate in this case is cell bank, and the sam-ple for testing before puriˆcation process is a harvested cellafter cell culturing or unprocessed bulk which consists ofsingle or pooled complex culture broth. The unprocessedbulk may be sometimes culture broth without cell. Denial oflatent virus, which is determined by virus test at a MCB orWCB level, does not always deny latent virus in unprocessedbulk after culturing. Further, it is noted that the viral test atthe CAL is meaningful as a validation but can not guaranteedeˆnite assurance of latent virus denial, since the test isgenerally performed only once. In case of using a serum or acomponent of blood origin in a culture medium, deˆnitedenial of latent virus at the level of unprocessed bulk can notbe assured so long as the viral test has not been conducted oneach lot at the CAL, since lot renewal can be a variable fac-tor on viral contamination.

A representative sample of the unprocessed bulk, removedfrom the production reactor prior to further processing,represents one of the most suitable levels at which the pos-sibility of adventitious virus contamination can be deter-mined with a high probability of detection. Appropriate test-ing for viruses should be performed at the unprocessed bulklevel unless virus testing is made more sensitive by initial par-tial processing (e.g., unprocessed bulk may be toxic in testcell cultures, whereas partially processed bulk may not betoxic). In certain instances it may be more appropriate to testa mixture consisting of both intact and disrupted cells andtheir cell culture supernatants removed from the productionreactor prior to further processing.

In case of unprocessed bulk, it is required to conduct virustest on at least 3 lots obtained from pilot scale or commercialscale production. It is recommended that manufacturersdevelop programs for the ongoing assessment of adventiti-ous viruses in production batches. The scope, extent and fre-quency of virus testing on the unprocessed bulk should bedetermined by taking several points into consideration in-cluding the nature of the cell lines used to produce thedesired products, the results and extent of virus tests per-formed during the qualiˆcation of the cell lines, the cultiva-tion method, raw material sources and results of viral clear-ance studies. Screening in vitro tests, using one or several celllines, are generally employed to test unprocessed bulk. If

appropriate, a NAT test or other suitable methods may beused.

Generally, harvest material in which adventitious virushas been detected should not be used to manufacture theproduct. If any adventitious viruses are detected at this level,the process should be carefully checked to determine thecause of the contamination, and appropriate actions taken.5.2 Virus test as an acceptance test of an intermediatematerial, etc.

When a biological product is manufactured from tissue,body ‰uid etc. of human or animal origin, there are casesthat an intermediate material, partially processed as a rawmaterial or substrate by outside manufacturer, is purchasedand used for manufacturing. In such case, if any test to meetthis General Information has been conducted by such out-side manufacturer, it is necessary for the manufacturer ofthe biological product, who purchased the intermediatematerial, to examine what sort of virus test has to be con-ducted as acceptance tests, and to keep record on the basis ofrationality including the details of the test conducted.

On the other hand, if no test to meet this General Infor-mation has been conducted by such outside manufacturer ofthe raw material, all necessary virus free test has to be con-ducted to meet this General Information on the intermediatematerial regarding it as the direct substrate for drug produc-tion.5.3 Virus test on a ˆnal product

Virus tests to be conducted on a ˆnal product (or on aproduct to reach the ˆnal product) has to be deˆned undercomprehensive consideration of the type of raw material orsubstrate, the result of virus test conducted on raw materialWsubstrate, the result of evaluation on viral removalWinactiva-tion process, any possibility of virus contamination in themanufacturing process, etc. Comprehensive viral safety as-surance can only be achieved by appropriate selection of theraw materialWsubstrate, an appropriate virus test conductedon the raw materialWsubstrateWintermediate material, thevirus test conducted at an appropriate stage of manufactur-ing, an appropriate viral clearance test, etc. However, thereare cases of having speciˆc backgrounds, such as 1) use ofthe raw material derived from unspeciˆed individual human,2) possible existence of virus at window period, 3) speciˆcdetection limit of virus test, etc. and in these cases, viruscontamination to the ˆnal product may occur if there is anydeˆciency on the manufacturing process (e.g., damage ofmembrane ˆlter) or any mix-up of the raw materials, etc. Toavoid such accidental virus contamination, it may be recom-mended to conduct nucleic ampliˆcation test (NAT) on theˆnal product focusing on the most risky virus among thosethat may possibly to exist in the raw material.

6. Process evaluation on viral clearance6.1 Rationale, objective and general items to be concernedwith respect to viral clearance process evaluation

Evaluation of a viral inactivationWremoval process is im-portant for ensuring safety of a biological product derivedfrom tissue or body ‰uid of human or animal origin. Con-ducting evaluation on viral clearance is to assure, even to

1627

Table 3. Example of viruses which have been used for viral clearance studies

Virus Family Genus Natural host Genome Env Size (nm) Shape Resistance

Vesicular Stomatitis Virus Rhabd Vesiculovirus EquineBovine

RNA yes 70× 150 Bullet Low

Parain‰uenza Virus Paramyxo Paramyxovirus Various RNA yes 100 – 200＋ Pleo-Spher Low

MuLV Retro Type Concovirus

Mouse RNA yes 80 – 110 Spherical Low

Sindbis Virus Toga Alphavirus Human RNA yes 60 – 70 Spherical Low

BVDV Flavi Pestivirus Bovine RNA yes 50 – 70 Pleo-Spher Low

Pseudorabies Virus Helpes Swine DNA yes 120 – 200 Spherical Med

Poliovirus Sabin Type 1 Picorna Enterovirus Human RNA no 25 – 30 Icosahedral Med

EncephalomyocardititisVirus

Picorna Cardiovirus Mouse RNA no 25 – 30 Icosahedral Med

Reovirus 3 Reo Orthoreovirus Various kind RNA no 60 – 80 Spherical Med

SV 40 Papova Polyomavirus Monkey DNA no 40 – 50 Icosahedral Very high

Parvovirus: canine, por-cine

Parvo Parvovirus CaninePorcine

DNA no 18 – 24 Icosahedral Very high


some extent, elimination of the virus, which may exist in araw material, etc. or may be derived to the process due tounexpected situation. Viral clearance studies should be madeby a carefully designed appropriate method, and has to berationally evaluated.

The objective of viral clearance studies is to assess processstep(s) that can be considered to be eŠective in inactivatingWremoving viruses and to estimate quantitatively the overalllevel of virus reduction obtained by the process. This shouldbe achieved by the deliberate addition (``spiking'') of sig-niˆcant amounts of a virus at diŠerent manufacturingWpuriˆcation steps and demonstrating its removal or inactiva-tion during the subsequent steps. It is not necessary to evalu-ate or characterize every step of a manufacturing process ifadequate clearance is demonstrated by the use of fewersteps. It should be borne in mind that other steps in the proc-ess may have an indirect eŠect on the viral inactivationWremoval achieved. Manufacturers should explain and justifythe approach used in studies for evaluating viral clearance.

The reduction of virus infectivity may be achieved byremoval of virus particles or by inactivation of viral infectiv-ity. For each production step assessed, the possible mechan-ism of loss of viral infectivity should be described withregard to whether it is due to inactivation or removal. Forinactivation steps, the study should be planned in such a waythat samples are taken at diŠerent times and an inactivationcurve constructed.6.2 Selection of virus

To obtain broad range of information of viral inactivationWremoval, it is desirable that a model virus used for viralclearance studies should be chosen from the viruses withbroad range of characteristics in aspects of DNAWRNA,with or without envelope, particle size, signiˆcant resistance

to physicalWchemical treatment, etc. and it is necessary tocombine about 3 model viruses to cover these characteristics.

At choice of a model virus, there are also the ways tochoose a virus closely related to or having the same charac-teristics of the virus known to exist in the raw material. Insuch case, it is in principle recommendable to choose a viruswhich demonstrates a higher resistance to inactivationWremoval treatment if two or more candidate viruses areavailable for choice. Further, a virus which can grow at ahigh titer is desirable for choice, although this may not al-ways be possible. In addition to the above, choosing a virus,which will provide eŠective and reliable assay result at eachstep, is necessary, since sample condition to be tested at eachstep of a production process may in‰uence the detection sen-sitivity. Consideration should also be given to health hazardwhich may pose to the personnel performing the clearancestudies.

For the other items taken for consideration at choice ofvirus, the Notice, Iyakushin No. 329 can be used as a refer-ence. Examples of the virus which have been used for viralclearance studies are shown in Table 3 which was derivedfrom Iyakushin No. 329. However, the Notice, IyakushinNo. 329, is on viral safety of a product derived cell line ofhuman or animal origin, and a more appropriate modelvirus has to be chosen taking into account the originWrawmaterial of biological products.6.3 Design of viral clearance studies

The purpose of viral clearance studies is to quantitativelyevaluate removal or inactivation capability of a process, inwhich a virus is intentionally spiked to a speciˆc step of amanufacturing process.

Following are the precautions to be taken at planning viralclearance studies.


(1) Care should be taken in preparing the high-titer virusto avoid aggregation which may enhance physical removaland decrease inactivation thus distorting the correlation withactual production.

(2) Virus detection method gives great in‰uence to viralclearance factor. Accordingly, it is advisable to gain detec-tion sensitivity of the methods available in advance, and usea method with a detection sensitivity as high as possible.Quantitative infectivity assays should have adequate sen-sitivity and reproducibility in each manufacturing process,and should be performed with su‹cient replicates to ensureadequate statistical validity of the result. Quantitative assaysnot associated with infectivity may be used if justiˆed. Ap-propriate virus controls should be included in all infectivityassays to ensure the sensitivity of the method. Also, thestatistics of sampling virus when at low concentrations (forexample, number of virus is 1-1000WL) should be considered.

(3) Viral clearance studies are performed in a miniaturesize system that simulates the actual production process ofthe biotechnologicalWbiological product used by themanufacturer. It is inappropriate to introduce any virus intoa production facility because of GMP constraints. There-fore, viral clearance studies should be conducted in aseparate laboratory equipped for virological work and per-formed by staŠ with virological expertise in conjunctionwith production personnel involved in designing and prepar-ing a scaled-down version of the puriˆcation process. Theviral clearance studies should be performed under the basicconcept of GLP.

(4) Each factor on a viral clearance study of a process,which is performed in miniature size, should re‰ect that ofactual manufacturing as far as possible, and its rationalityshould be clariˆed. In case of chromatograph process,length of column bed, linear velocity, ratio of bed volumeper velocity (in other words, contact time), buŠer, type ofcolumn packing, pH, temperature, protein concentration,salt concentration and concentration of the objectiveproduct are all correspondent to those of the actual produc-tion. Further, similarity of elution proˆle should beachieved. For the other process, similar concept should beapplied. If there is any factor which can not re‰ect the actualproduction, its eŠect to the result should be examined.

(5) It is desirable that two or more inactivationWremovalprocesses of diŠerent principles are selected and examined.

(6) As for the process which is expected to inactivateWre-move virus, each step should be evaluated in aspect of clear-ance capability, and carefully determined if it is the stage ofinactivation, removal or their combination for designing thetest. Generally, in viral clearance test, a virus is spiked ineach step which is the object of the test, and after passingthrough the process in question, the reduction level of infec-tivity is evaluated. But, in some case, it is accepted that ahigh potential virus is spiked at a step of the process, andvirus concentration of each succeeding step is carefullymonitored. When removal of virus is made by separation orfractionation, it is desirable to investigate how the virus isseparated or fractionated (mass balance).

(7) For assessment of viral inactivation, unprocessed

crude material or intermediate material should be spikedwith infectious virus and the reduction factor calculated. Itshould be recognized that virus inactivation is not a simple,ˆrst order reaction and is usually more complex, with a fast``phase 1'' and a slow ``phase 2''. The study should, there-fore, be planned in such a way that samples are taken atdiŠerent times and an inactivation curve constructed. It isrecommended that studies for inactivation include at leastone time point less than the minimum exposure time andgreater than zero, in addition to the minimum exposuretime. The reproducible clearance should be demonstrated inat least two independent studies. When there is a possibilitythat the virus is a human pathogen, it is very important thatthe eŠective inactivation process is designed and additionaldata are obtained. The initial virus load should be deter-mined from the virus which can be detected in the spikedstarting material. If this is not possible, the initial virus loadmay be calculated from the titer of the spiking virus prepara-tion. Where inactivation is too rapid to plot an inactivationcurve using process conditions, appropriate controls shouldbe performed to demonstrate that infectivity is indeed lostby inactivation.

(8) If antibody against virus exists in an unprocessedmaterial, caution should be taken at clearance studies, sinceit may aŠect the behavior of virus at viral removal or inacti-vation process.

(9) Virus spiked in unprocessed material should besu‹cient enough to evaluate viral removal or inactivationcapability of the process. However, the virus ``spike'' to beadded to the unprocessed material should be as small as pos-sible in comparison with the sample volume of the unproc-essed material so as not to cause characteristic change of thematerial by addition of the virus nor to cause behavioralchange of the protein in the material.

(10) It is desirable that the virus in the sample is subjectfor quantitative determination without applying ultracen-trifuge, dialysis, storage, etc. as far as possible. However,there may be a case that any handling before quantitativetest, such as remove procedure of inhibitor or toxic sub-stance, storage for a period to realize test at a time, etc., isinevitable. If any manipulation, such as dilution, concentra-tion, ˆltration, dialysis, storage, etc., is applied for prepara-tion of the sample for testing, a parallel control test, whichpasses through a similar manipulation, should be conductedto assess infectivity variance at the manipulation.

(11) BuŠers and product (desired protein or other com-ponent contained therein) should be evaluated independent-ly for toxicity or interference in assays used to determine thevirus titer, as these components may adversely aŠect the in-dicator cells. If the solutions are toxic to the indicator cells,dilution, adjustment of the pH, or dialysis of the buŠer con-taining spiked virus might be necessary. If the product itselfhas anti-viral activity, the clearance study may need to beperformed without the product in a ``mock'' run, althoughomitting the product or substituting a similar protein thatdoes not have anti-viral activity could aŠect the behaviour ofthe virus in some production steps.

(12) Many puriˆcation schemes use the same or similar


buŠers or columns, repetitively. The eŠects of this approachshould be taken into account when analyzing the data. TheeŠectiveness of virus elimination by a particular process mayvary with the stage in manufacture at which it is used.

(13) Overall reduction factors may be underestimatedwhere production conditions or buŠers are too cytotoxic orvirucidal and should be discussed on a case-by-case basis.Overall reduction factors may also be overestimated due toinherent limitations or inadequate design of viral clearancestudies.

(14) It has to be noted that clearance capability of viralremovalWinactivation process may vary depending upon thetype of virus. The viral removalWinactivation process, whichdisplays viral clearance by speciˆc principle or mechanism,may be quite eŠective to the virus, which meets suchmechanism of action, but not eŠective to the other type ofviruses. For example, SWD treatment is generally eŠective tothe virus with lipid membrane, but not eŠective to the virusnot having such membrane. Further, some virus is resistantto the general heating process (55 – 609C, 30 minutes).When clearance is expected for such virus, introduction of afurther severe condition or process, which has diŠerent prin-ciple or mechanism, is necessary. Virus removal membraneˆltration, which is diŠerent from SWD or heat treatment inaspect of principle, is eŠective to a broad range of virus thatcan not pass through the membrane. A‹nity chro-matography process, which speciˆcally absorbs the objectiveprotein, can thoroughly wash out the materials other thanthe objective protein including virus etc. and is generallyeŠective for viral removal. SeparationWfractionation of avirus from an objective protein is sometimes very di‹cult,but there are not so rare that ion exchange chromatography,ethanol fractionation, etc. is eŠective for clearance of a viruswhich can not be su‹ciently inactivated or removed by theother process.

(15) EŠective clearance may be achieved by any of thefollowing: multiple inactivation steps, multiple complemen-tary separation steps, or combinations of inactivation andseparation steps. Separation methods may be dependent onthe extremely speciˆc physico-chemical properties of a viruswhich in‰uence its interaction with gel matrices and precipi-tation properties. However, despite these potential varia-bles, eŠective removal can be obtained by a combination ofcomplementary separation steps or combinations of inacti-vation and separation steps. Well designed separation steps,such as chromatographic procedures, ˆltration steps andextractions, can be also eŠective virus removal steps pro-vided that they are performed under appropriately con-trolled conditions.

(16) An eŠective virus removal step should givereproducible reduction of virus load shown by at least twoindependent studies.

(17) Over time and after repeated use, the ability ofchromatography columns and other devices used in thepuriˆcation scheme to clear virus may vary. Some estimateof the stability of the viral clearance after several uses mayprovide support for repeated use of such columns.

(18) The Notice, Iyakushin No. 329, would be used as a

reference when viral clearance studies on biological productsare designed.6.4 Interpretation of viral clearance studies6.4.1 Evaluation on viral clearance factor

Viral clearance factor is a logarithm of reduction ratio ofviral amount (infectious titer) between each step applied forviral clearance of a manufacturing process. Total viral clear-ance factor throughout the process is sum of the viral clear-ance factor of each step appropriately evaluated.

Whether each and total viral clearance factor obtained areacceptable or should not be evaluated in aspects of everyvirus that can be realistically anticipated to derive into theraw material or the manufacturing process, and its rationali-ty should be recorded.

In case that existence of any viral particle is recognized ina substrate for drug production, e.g., a substrate of rodentorigin for biodrug production, it is important not only todemonstrate removal or inactivation of such virus, but alsoto demonstrate that the puriˆcation process has enoughcapability over the required level to assure safety of the ˆnalproduct at an appropriate level. The virus amount removedor inactivated in a manufacturing process should be com-pared with the virus amount assumed to exist in the substrateetc. used for manufacturing drug, and for this purpose, it isnecessary to obtain the virus amount in the raw materialsWsubstrate, etc. Such ˆgure can be obtained by measuring in-fectious titer or by the other method such as transmissionelectron microscope (TEM). For evaluation of overall proc-ess, a virus amount, far larger than that assumed to exist inthe amount of the raw materialsWsubstrate which is equiva-lent to single administration of the ˆnal product, has to beremoved. It is quite rare that existence of virus can be as-sumed in a substrate for drug production, with the exceptionof the substrate of rodent origin, and such suspicious rawmaterialWsubstrate should not be used for manufacturingdrug with a special exceptional case that the drug in questionis not available from the other process and is clinically in-dispensable, and that the information including infectiousproperties of the virus particle assumed to exist has beenclariˆed.6.4.2 Calculation of viral clearance index

Viral clearance factor, ``R'', for viral removalWinactiva-tion process can be calculated by the following formula.

R＝ log[(V1× T1)W(V2× T2)]

In whichR: Logarithm of reduction ratioV1: Sample volume of the unprocessed materialT1: Virus amount (titer) of the unprocessed materialV2: Sample volume of the processed materialT2: Virus amount (titer) of the processed material

At the calculation of viral clearance factor, it is recom-mendable to use the virus titer detected in the sample prepa-ration of the unprocessed material after addition of virus,not the viral titer added to the sample preparation whereverpossible. If this is not possible, loaded virus amount is calcu-lated from virus titer of the solution used for spike.


6.4.3 Interpretation of results and items to be concerned atevaluation

At the interpretation and the evaluation of the data oneŠectiveness of viral inactivationWremoval process, there arevarious factors to be comprehensively taken into account,such as appropriateness of the virus used for the test, design of the viral clearance studies, virus reduction ratioshown in logarithm, time dependence of inactivation, factorsWitems which give in‰uence to the inactivationWremoval process, sensitivity limit of virus assay method, possible eŠect of the inactivationWremoval process whichis speciˆc to certain class of viruses.

Additional items to be concerned at appropriate interpre-tation and evaluation of the viral clearance data are as fol-lows:(1) Behavior of virus used to the test

At interpretation of the vial clearance results, it is necessa-ry to recognize that clearance mechanism may diŠer depend-ing upon the virus used for the test. Virus used for a test isgenerally produced in tissue culture, but behavior of thevirus prepared in the tissue culture may be diŠerent fromthat of the native virus. Examples are possible diŠerences ofpurity and degree of aggregation between the native and thecultured viruses. Further, change of surface properties of avirus, e.g., addition of a sucrose chain which is ascribed tospeciˆc nature of a separation process, may give eŠect to theseparation. These matters should be also considered at inter-pretation of the results.(2) Design of test

Viral clearance test should have been designed taking intoaccount variation factors of the manufacturing process andscaling down, but there still remain some variance fromactual production scale. It is necessary to consider such vari-ance at the interpretation of the data and limitation of thetest.(3) Acceptability of viral reduction data

Total viral clearance factor is expressed as a sum oflogarithm of reduction ratio obtained at each step. The sum-mation of the reduction factor of multiple steps, particularlyof steps with little reduction (e.g., below 1 log10), mayoverestimate viral removalWinactivation capability of theoverall process. Therefore, virus titer of the order of 1 log10

or less has to be ignored unless justiˆed. Further, viral clear-ance factor achieved by repeated use of the same or similarmethod should be ignored for calculation unless justiˆed.(4) Time dependence of inactivation

Inactivation of virus infectivity frequently shows biphasiccurve, which consists of a rapid initial phase and subsequentslow phase. It is possible that a virus not inactivated in a stepmay be more resistant to the subsequent step. For example,if an inactivated virus forms coagulation, it may be resistantto any chemical treatment and heating.(5) Evaluation of viral reduction ratio shown logarithm

Viral clearance factor shown in logarithm of reductionratio of virus titer can demonstrate drastic reduction ofresidual infectious virus, but there is a limit that infectioustiter can never be reduced to zero. For example, reduction ininfectivity of a preparation containing 8 log10 infectious unit

per mL by a factor of 8 log10 leaves zero log10 per mL or oneinfectious unit per mL, taking into account the detectionlimit of assay.(6) Variable factor of manufacturing process

Minor variance of a variation factor of a manufacturingprocess, e.g., contact time of a spiked sample to a buŠer or acolumn, will sometimes give in‰uence to viral removal or in-activation eŠect. In such case, it may be necessary to inves-tigate to what extent such variance of the factor has given in-‰uence to the process concerned in aspect of viral inactiva-tion.(7) Existence of anti-viral antiserum

Anti-viral antiserum that exists in the sample preparationused for a test may aŠect sensitivity of distribution or inacti-vation of a virus, which may result in not only defusing thevirus titer but complicating interpretation of the test result.So, existence of anti-viral antiserum is one of the importantvariable factors.(8) Introduction of a new process for removalWinactivation

Viral clearance is an important factor for securing safetyof drug. In case that an achievement level of infective clear-ance of a process is considered insu‹cient, a process whichis characterized by inactivationWremoval mechanism to meetthe purpose or an inactivationWremoval process which canmutually complement to the existence process has to be in-troduced.(9) Limit of viral clearance studies

Viral clearance studies are useful for contributing to theassurance that an acceptable level of safety in the ˆnalproduct is achieved but do not by themselves establish safe-ty. However, a number of factors in the design and execu-tion of viral clearance studies may lead to an incorrect esti-mate of the ability of the process to remove virus infectivity,as described above.

7. StatisticsThe viral clearance studies should include the use of

statistical analysis of the data to evaluate the results. Thestudy results should be statistically valid to support the con-clusions reached.7.1 Statistical considerations for assessing virus assays

Virus titrations suŠer the problems of variation commonto all biological assay systems. Assessment of the accuracyof the virus titrations and reduction factors derived fromthem and the validity of the assays should be performed todeˆne the reliability of a study. The objective of statisticalevaluation is to establish that the study has been carried outto an acceptable level of virological competence.Assay

1. Assay methods may be either quantal or quantitative.Both quantal and quantitative assays are amenable tostatistical evaluation.

2. Variation can arise within an assay as a result of dilu-tion errors, statistical eŠects and diŠerences within the assaysystem which are either unknown or di‹cult to control.These eŠects are likely to be greater when diŠerent assayruns are compared (between-assay variation) than whenresults within a single assay run are compared (within-assay


variation).3. The 95z conˆdence limits for results of within-assay

variation normally should be on the order of ±0.5 log10 ofthe mean. Within-assay variation can be assessed by stan-dard textbook methods. Between-assay variation can bemonitored by the inclusion of a reference preparation, theestimate of whose potency should be within approximately0.5 log10 of the mean estimate established in the laboratoryfor the assay to be acceptable. Assays with lower precisionmay be acceptable with appropriate justiˆcation.7.2 Reproducibility and conˆdence limit of viral clearancestudies

An eŠective virus inactivationWremoval step should givereproducible reduction of virus load shown by at least twoindependent studies. The 95z conˆdence limits for thereduction factor observed should be calculated whereverpossible in studies of viral clearance. If the 95z conˆdencelimits for the viral assays of the starting material are±s, andfor the viral assays of the material after the step are±a, the95z conˆdence limits for the reduction factor are± s2＋ a2.

8. Re-evaluation of viral clearanceWhenever signiˆcant changes in the production or puriˆ-

cation process are made, the eŠect of that change, bothdirect and indirect, on viral clearance should be consideredand the system re-evaluated as needed. Changes in processsteps may also change the extent of viral clearance.

9. Measurement for viral clearance studies9.1 Measurement of virus infective titer

Assay methods may be either quantal or quantitative.Quantal methods include infectivity assays in animals or intissue-culture-infectious-dose (TCID) assays, in which theanimal or cell culture is scored as either infected or not. In-fectivity titers are then measured by the proportion ofanimals or culture infected. In quantitative methods, the in-fectivity measured varies continuously with the virus input.Quantitative methods include plaque assays where each p-laque counted corresponds to a single infectious unit. Bothquantal and quantitative assays are amenable to statisticalevaluation.9.2 Testing by nucleic-acid-ampliˆcation test (NAT)

NAT can detect individual or pooled raw materialWcellsubstrate or virus genome at a high sensitivity even in a stagethat serum test on each virus is negative. Further, it can de-tect HBV or HCV gene, which can not be measured in cul-ture system. Window period can be drastically shortened atthe test on HBV, HCV and HIV, and the method is expectedto contribute as an eŠective measure for ensuring viral safe-ty. However, depending upon a choice of primer, there maybe a case that not all the subtype of objective virus can be de-tected by this method, and, therefore, it is recommendableto evaluate, in advance, if subtype of a broad range can bedetected.

NAT will be an eŠective evaluation method for virusremoval capability at viral clearance studies. However, incase of viral inactivation process, viral inactivation obtainedby this method may be underrated, since there is a case that

inactivated virus still shows positive on nucleic acid. Fur-ther, at introduction of NAT, cautions should be taken onrationality of detection sensitivity, choice of a standardwhich is used as run-control, quality assurance and main-tenance of a reagent used for primer, interpretation of posi-tive and negative results, etc.

10. Reporting and preservationAll the items relating to virus test and viral clearance stu-

dies should be reported and preserved.

11. OthersThe Notice, Iyakushin No. 329, should be used as a refer-

ence at virus test and viral clearance studies.

ConclusionAs mentioned at the Introduction, assurance of qualityW

safety etc. of JP listed drugs should be achieved by state-of-the-art methods and concepts re‰ecting the progress ofscience and accumulation of experiences.

The basis for ensuring viral safety of JP listed biotechno-logicalWbiological products is detailed in this General Infor-mation. What is discussed here is that an almost equal levelof measures are required for both development of new drugsand for existing products as well, which means that similarlevel of concerns should be paid on both existing and newproducts in aspect of viral safety. This document is intendedto introduce a basic concept that quality and safety assur-ance of JP listed product should be based upon the most ad-vanced methods and concepts. This document has been writ-ten to cover all conceivable cases, which can be applied to allbiotechnologicalWbiological products. Therefore, there maybe cases that it is not so rational to pursue virus tests andviral clearance studies in accordance with this document oneach product, which has been used for a long time withoutany safety issue. So, it will be necessary to elaborate themost rational ways under a case-by-case principle taking intodue consideration source, origin, type, manufacturing proc-ess, characteristics, usages at clinical stage, accumulation ofthe past usage record, etc. relating to such biotechnologicalWbiological products.

Add the following:

18. Qualiˆcation of Animals asOrigin of Animal-derived MedicinalProducts provided in the General

Notices 39 of JapanesePharmacopoeia and

Other StandardsIntroduction

The O‹cial Gazette issued on March 29, 2002 announcedthat General Notices or General Rules of the Japanese Phar-macopoeia of the Standard of Drugs not required Approval,of the Biological Products Standard and of the Require-


ments for Antibiotic Products of Japan were amended toadd a provision that ``When a drug product or a drug sub-stance which is used to manufacture a drug product, ismanufactured from a raw material of animal origin, theanimal in question should be in principle a healthy subject, ifnot otherwise provided.''.

The Notice Iyaku-hatsu No. 0329001, which was issued onthe same date, provided that ``Healthy subject herein pro-vided is the animal which does not cause any disease or anyinfection to human being at an appropriate production proc-ess and use of the drug product, and as for the oral or exter-nal drug for example, the animal, as its raw material ofanimal origin, should be conˆrmed at this stage to meet theFood Standard. It has to be noted that this standard ofhealthy subject has to be revised timely taking into accountthe up-to-date information with respect to the amphixenosisinfections common between human beings and animals.''.

This General Information describes safety assuranceagainst infection of drugs, which are manufactured fromraw materials of animal origin, to follow up the Notice asmentioned above.

1. Basic conceptWhen drugs derived from raw materials of animal origin

including human are used, it is important to take into ac-count any possibility that communicable disease agents suchas virus may cause infectious disease or any possible hazardsto patients. In such case, it goes without saying that the pri-mary subject that has to be considered is the absence of anyinfectious agents such as virus in the raw materials of animalorigin including human as the source of the drug. More im-portant points are whether the drugs derived from such rawmaterials are free of such infectious agents and whetherthere is any possibility of transmission of infectious agentswhen the drugs are administered to patient. The eligibility ofanimals including human, as the source of raw materials ofdrugs, in other words ``the subject which is free from anydisease or transmission of infectious agents that is infectiousto human being at an appropriate production process anduse of the drug product'' is that ``The drug should be entire-ly free from any risk of infections by means of whole proce-dures which include evaluation of appropriateness of theanimals including human as the source of their raw materi-als, establishment of appropriate production processes andtheir appropriate control, and strict adherence to the clinicalindications of the ˆnal product.''

2. Animals including human as the source of raw materialsof drugs

What is the most clear and appropriate preventive meas-ures against infection to human being due to administrationof drugs which are derived from animals including human isto assure the absence of any infectious agents such as virus inits raw materials or an appropriate critical raw material byeach of the followings: (1) the use of raw materials ofhealthy animal origin, which are proved to be free fromcommunicable disease agents to human, or (2) the use of ap-propriate critical raw materials (e.g., cell substrate, bloodplasma, pooled urine after some treatments) for drug

production, which are proved to be free from communicabledisease agents after certain appropriate processing on rawmaterials of animal origin.

As for raw materials of drugs of human origin, cell, tis-sue, blood, placenta, urine, etc. are used. Whenever it issu‹cient and possible each donor, as the origin of such rawmaterials, should be asked his (her) health condition and un-dergoes his (her) medical examination at this stage, so thatthe appropriateness as a donor can be conˆrmed from thestandpoint of safety concerning communicable diseaseagents such as virus.

For example, ``Basic concept on handling and use of adrug product, etc. which is derived from cellWtissue''(Attachment 1 of the Notice Iyaku-Hatsu No. 1314 datedDecember 26, 2000) and ``Guidance for quality and safetyassurance of a drug product, etc. which is derived from hu-man cellWtissue (Attachment 2 of the Notice Iyaku-HatsuNo. 1314 dated December 26, 2000)'' issued by the Director-General of the Medicinal Safety Bureau, Ministry of Healthand Welfare, states that since the cellWtissue supplied by ahuman donor comes to be applied to patients without proc-essing through any su‹cient inactivation or removal of com-municable disease agents, the selection and qualiˆcationcriteria on such donor has to be established. These criteriaare to be composed with the respect to the check items on thecase history and the physical conditions as well as the testitems on the various transmission of infectious agentsthrough cellWtissue, and that the appropriateness of thesecriteria has to be clariˆed. Hepatitis Type-B (HBV), Hepati-tis Type-C (HCV), Human Immune Deˆciency Viral infec-tions (HIV), Adult T-Cell Leukemia and Parvovirus B19 In-fections should be denied through the interview to the donorand the tests (serologic test, nucleic-acid ampliˆcation test,etc.). Further, if necessary, Cytomegalovirus infection andEB Virus infection should be denied by tests. ``Infectionscaused by bacteria such as Treponema pallidum,Chlamydia, Gonococci, Tubercule bacillus, etc.'', ``septice-mia and its suspicious case'', ``vicious tumor'', ``seriousmetabolic or endocrine-related disorders'', ``collagenosisand haematological disorder'', ``hepatic disease'' and ``de-mentia (transmissible spongiform encephalopathies and itssuspicious case)'' should be checked on the case history orby the interview, etc. and the experience of being transfusedorWand transplanted should be checked to conˆrm eligibilityas a donor. The most appropriate check items and testmethods then available are to be used, which need to bereconsidered at appropriate timing taking into account theupdated knowledge and the progress of the science and thetechnologies. At screening of a donor, reexaminations has tobe made at appropriate timing using the eligible check itemsand the test methods taking into account the window period(Initial period after infection, in which antibody against bac-teria, fungi or virus is not detected.)

In the case of plasma derivatives produced from the do-nated blood in Japan, the donor should be checked bymeans of self-assessed report about health conditions, and aserologic check and a nucleic acid ampliˆcation test (NAT)on 50 pooled plasma should be performed at the stage of do-


nated blood. Further, the plasma material (i.e., critical rawmaterial ) for fractionation should be stored 4 months inminimum so that the arrangement could be taken based onthe information available after collection of the blood andthe blood infusion to exclude the possibility of using anycritical raw material which might cause infection to patients.

On the other hand, as for the materials such as urinewhich are taken from the unspeciêd number of the donorsand come to be critical raw materials for drug productionafter some treatments, it is unrealistic and not practical toconduct the tests of virus infection, etc. on the individualdonor. Consequently, appropriate tests such as virus test hasto be performed on such critical raw materials for drugproduction.

In the case of the animals besides human, the wild onesshould be excluded. Only the animals, which are raised un-der well sanitarily controlled conditions taken to preventbacterial contamination or under the eŠective bacterial pol-lution monitoring systems, have to be used, and it is recom-mended that the animals from a colony appropriately con-trolled under speciˆc pathogen-free (SPF) environment areto be used as far as possible. Further, for the animals regu-lated under the Food Standard, only the animals that metthis standard should be used. It should be conˆrmed by ap-propriate tests that the animals were free from pathogen, ifnecessary.

The concrete measures to avoid transmittance or spread ofinfectivity of prion, which is considered to be the pathogenof transmissible spongiform encephalopathies (TSEs), as faras possible are the followings: avoidance of use ofanimals, which are raised in the areas where high incidenceor high risk of TSEs (Scrapie in sheep and goat, bovinespongiform encephalopathies (BSE) in cattle, chronic wast-ing disease (CWD) in deer, new type of Creutzfeldt-Jacob-Disease (CJD) in human, etc.) is reported, and humans, whohave stayed long time (more than 6 months) in such areas, asraw materials or related substances of drugs; avoidanceof use of any substances that are derived from the individualinfected with scrapie, BSE, CJD, etc.; avoidance of usinga material derived from organ, tissue and cell, etc. of highrisk of TSEs; and taking appropriate measures basing onthe information collected, which includes incidence of TSEs,the results of epidemiological investigation and the ex-perimental research on prion, and incidence of tardive infec-tion on donors after collecting raw materials, etc.

3. Human or animal cells which are used as critical rawmaterials for drug production

Cell substrates derived from humans or animals are usedfor drug production. In such case, it is desirable that thehumans or the animals, which are the origins of the cell sub-strates, are healthy subjects. However, it is considered prac-tical that viral safety of the drugs derived from the cell sub-strates are evaluated on the cells, which are so called criticalraw materials for production of such drugs. In such case, thesafety should be conˆrmed through the test and analysis onestablished cell bank thoroughly with respect to virus etc., asfar as possible. The items and the methods of the tests that

have been followed in this case are described in detail in theNotice of Japanese version on the internationally acceptedICH Guideline entitled ``Viral safety evaluation ofbiotechnology products derived from cell lines of human oranimal origin'' (Iyakushin No. 329 issued on February 22,2000 by Director, Evaluation and Licensing Division, Phar-maceutical and Medical Safety Bureau, Ministry of Healthand Welfare). In the meantime, it is important how to han-dle the cell in case that any virus has been detected under thecell level tests. This Notice describes how to cope with thissituation as follows: `Ìt is recognised that some cell linesused for the manufacture of product will contain en-dogenous retroviruses, other viruses or viral sequences. Insuch circumstances, the action plan recommended formanufacturer is described in Section V (Rationale and actionplan for viral clearance studies and virus tests on puriêdbulk) of the Notice. The acceptability of cell lines containingviruses other than endogenous retroviruses will be consi-dered on an individual basis by the regulatory authorities, bytaking into account a riskWbeneˆt analysis based on thebeneˆt of the product and its intended clinical use, thenature of the contaminating viruses, their potential forinfecting humans or for causing disease in humans, thepuriˆcation process for the product (e.g., viral clearanceevaluation data), and the extent of the virus tests conductedon the puriêd bulk.'' For example, it is well known thatType A-, R- and C-endogenous particles like retrovirus areobserved in the cells of the rodents used most often for drugproduction. It is also known that they are not infectious tohuman and is not dangerous, and CHO cells are generallyused for drug production. The established cell lines (e.g.,NAMALWA Cell, BALL-1 Cell, etc.) derived from cancerpatients are sometimes used, but through the thorough virustests, etc., their safety are conˆrmed. The established celllines are assumed to be safer than the primary cultured cellswhich are hard to conduct the thorough virus test.

4. Establishment and control of appropriate productionprocess and adherence to the clinical indication of ˆnalproduct for safety assurance

Safety assurance against potential infections at only thelevel of animals that are source of raw materials of drugs islimited. Further, ``health of animal'' can not be deˆnedunivocally, and the various factors have to be taken into ac-count. The ˆnal goal of this subject is to protect humanfrom any infectious disease caused by drugs. Achieving thisgoal, the establishment and control of appropriate produc-tion processes of each drug and the adherence to the clinicalindications of the ˆnal product are important.

As mentioned above, the rodent cells used most often forthe production of the drugs are known to have endogenousretrovirus sometimes. The reason why such cells can be usedfor the production of the drugs is that multiple measures areapplied for safety in the puriˆcation stages which include ap-propriate inactivation or removal processes. There are casesin which the production procedure involves intentional useof a virus or a microorganism. In this case, relevant meas-ures capable of removing or inactivating of such virus or


microorganism are appropriately incorporated in the puriˆ-cation process, so that the risk of infection to human can befully denied and its safety can be assured when it is used as adrug. Further, even in the case that it is di‹cult to clarify therisk of contamination of the infectious agents or that the rawmaterial are contaminated by viruses etc., the raw materialin question may be used for the production of drugs so longas appropriate inactivation or removal processes are in-troduced, their eŠectiveness can be conˆrmed and the safetycan be assured by appropriate control of the manufacturingprocesses under GMP, etc.

5. ConclusionThe qualiˆcation of animals including human, as the

source of raw materials of drugs, in other words ``the sub-ject which does not cause any infectious diseases to humanbeing at an appropriate production process and use of thedrug product'' is that ``the drug has to be entirely free fromany risk of infections by means of whole procedures whichinclude evaluation of appropriateness of the animal includ-ing human as the source of their raw materials, establish-ment of appropriate production processes and theirappropriate control, and strict adherence to the clinical indi-cation of the ˆnal product.''

To cope with this subject, the advanced scientiˆc meas-ures, which actually re‰ect the updated knowledge andprogress of the science and the technology about infectiousdiseases in human and infection of animal origin, have to betaken into account timely.

Add the following:

19. SDS-Polyacrylamide GelElectrophoresis

The SDS-Polyacrylamide Gel Electrophoresis is used forthe characterization of proteins in biotechnological and bio-logical products and for control of purity and quantitativedeterminations.

This technique is a suitable analytical method with whichto identify and to assess the homogeneity of proteins inbiotechnological and biological products. The method isalso routinely used for the estimation of protein subunitmolecular masses and for determining the subunit composi-tions of puriˆed proteins.

Ready-to-use gels and reagents are widely available on themarket and can be used instead of those described in thistext, provided that they give equivalent results and that theymeet the validity requirements given below under Validationof the Test.

1. Characteristics of Polyacrylamide GelsThe sieving properties of polyacrylamide gels are aŠorded

by the three-dimensional network of ˆbers and pores whichis formed as the bifunctional bisacrylamide cross-links adja-cent polyacrylamide chains. Polymerization is catalyzed by afree radical-generating system composed of ammonium

persulfate and N,N,N?,N?-tetramethylethylenediamine(TEMED).

As the acrylamide concentration of a gel increases, itseŠective pore size decreases. The eŠective pore size of a gel isoperationally deˆned by its sieving properties; that is, by theresistance it imparts to the migration of macromolecules.There are limits on the acrylamide concentration that can beused. At high acrylamide concentrations, gels break muchmore easily and are di‹cult to handle. As the pore size of agel decreases, the migration rate of a protein through the geldecreases. By adjusting the pore size of a gel, throughmanipulating the acrylamide concentration, the resolutionof the method can be optimized for a given protein product.Thus, the physical characteristics of a given gel are deter-mined by the relative concentrations of acrylamide and bi-sacrylamide, used in its preparation.

In addition to the composition of the gel, the state of theprotein is an important determinant of the electrophoreticmobility. In the case of proteins, the electrophoretic mobili-ty is dependent on the pK values of the charged groups andthe size of the molecule. It is also in‰uenced by the type,concentration and pH of the buŠer, the temperature and theˆeld strength, as well as by the nature of the supportmaterial.

2. Polyacrylamide Gel Electrophoresis under DenaturingConditions

The method cited as an example is limited to the analysisof monomeric polypeptides with a mass range of 14,000 to100,000 daltons. It is possible to extend this mass range byvarious techniques (e.g., by using gradient gels, particularbuŠer systems, etc.), but those techniques are not discussedin this chapter.

Analysis by electrophoresis on sodium dodecyl sulfate(SDS) polyacrylamide gel (SDS-Polyacrylamide Gel Elec-trophoresis) under denaturing conditions is the most com-mon mode of electrophoresis used in assessing the quality ofproteins in biotechnological and biological products, andwill be the focus of the example described here. Typically,analytical electrophoresis of proteins is carried out in poly-acrylamide gels under conditions that ensure dissociation ofthe proteins into their individual polypeptide subunits andthat minimize aggregation. Most commonly, the stronglyanionic detergent SDS is used in combination with heat todissociate the proteins before they are loaded on the gel. Thedenatured polypeptides bind to SDS, become negativelycharged and exhibit a consistent charge-to-mass ratioregardless of protein type. Because the amount of SDSbound is almost always proportional to the molecular massof the polypeptide and is independent of its amino acid se-quence, SDS-polypeptide complexes migrate through poly-acrylamide gels with mobilities that are dependent on thesize of the polypeptides.

The electrophoretic mobilities of the resultant SDS-polypeptide complexes all assume the same functionalrelationship to their molecular masses. Migration of SDS-complexes occurs toward the anode in a predictable manner,with low-molecular-mass complexes migrating faster than


larger ones. The molecular mass of a protein can thereforebe estimated from its relative mobility calibrated in SDS-Polyacrylamide Gel Electrophoresis and the occurrence of asingle band in such a gel is a criterion of purity.

However, modiˆcations to the polypeptide backbone,such as N- or O-1inked glycosylation, have a signiˆcant im-pact on the apparent molecular mass of a protein, since SDSdoes not bind to a carbohydrate moiety in a manner similarto a polypeptide. Thus, a consistent charge-to-mass ratio isnot maintained. The apparent molecular masses of proteinsthat have undergone post-translational modiˆcations do nottruly re‰ect the masses of the polypeptides.1) Reducing conditions

Polypeptide subunits and three-dimensional structure ofproteins are often ˆxed, at least in part, by the presence ofdisulˆde bonds. A goal of SDS-Polyacrylamide Gel Elec-trophoresis under reducing conditions is to disrupt thisstructure by reducing the disulˆde bonds. Complete denatu-ration and dissociation of proteins by treatment with 2-mer-captoethanol or dithiothreitol (DTT) will result in unfoldingof the polypeptide backbone and subsequent complexationwith SDS. Under these conditions, the molecular masses ofthe polypeptide subunits can be calculated by interpolationin the presence of suitable molecular-mass standards.2) Non-reducing conditions

For some analyses, complete dissociation of the protein ofinterest into subunit peptides is not desirable. In the absenceof treatment with reducing agents such as 2-mercap-toethanol or DTT, disulˆde covalent bonds remain intact,preserving the oligomeric form of the protein. OligomericSDS-protein complexes migrate more slowly than their SDS-polypeptide subunits. In addition, non-reduced proteinsmay not be completely saturated with SDS and, hence, maynot bind the detergent in the expected mass ratio. Thismakes molecular-mass determinations of these molecules bySDS-Polyacrylamide Gel Electrophoresis less straightfor-ward than analyses of fully denatured polypeptides, since itis necessary that both standards and unknown proteins be insimilar conˆgurations for valid comparisons. However, thestaining of a single band in such a gel is a criterion of purity.

3. Characteristics of Discontinuous BuŠer System GelElectrophoresis

The most widely used electrophoretic method for the anal-ysis of complex mixtures of proteins involves the use of adiscontinuous buŠer system consisting of two contiguous,but distinct gels: a resolving or separating (lower) gel and astacking (upper) gel. The two gels are cast with diŠerentporosities, pH, and ionic strengths. In addition, diŠerentmobile ions are used in the gel and electrode buŠers. ThebuŠer discontinuity acts to concentrate large-volume sam-ples in the stacking gel, resulting in improved resolution.When power is applied, a voltage drop develops across thesample solution which drives the proteins into the stackinggel. Glycinate ions from the electrode buŠer follow the pro-teins into the stacking gel. A moving boundary region is rap-idly formed with the highly mobile chloride ions in the frontand the relatively slow glycinate ions in the rear. A localized

high-voltage gradient forms between the leading and trailingion fronts, causing the SDS-protein complex to form into avery thin zone (called the stack) and to migrate between thechloride and glycinate phases. Regardless of the height ofthe applied sample solution in the wells, all SDS-proteincomplexes condense within broad limits and enter the resolv-ing gel as a well-deˆned, thin zone of high protein density.The large-pore stacking gel does not retard the migration ofmost proteins and serves mainly as an anticonvective medi-um. At the interface of the stacking and resolving gels, theproteins experience a sharp increase in retardation due to thesmaller pore size of the resolving gel. Once the proteins arein the resolving gel, their mobility continues to be sloweddown by the molecular sieving eŠect of the matrix. Theglycinate ions overtake the proteins, which then move in aspace of uniform pH formed by the tris(hydrox-ymethyl)aminomethane and glycine. Molecular sievingcauses the SDS-polypeptide complexes to separate on thebasis of their molecular masses.

4. Preparing Vertical Discontinuous BuŠer SDS-Polyacrylamide Gels1) Assembling of the gel moulding cassette

Clean the two glass plates (size: e.g. 10 cm× 8 cm), thesample comb made of polytetra‰uoroethylene, the twospacers and the silicone rubber tubing (diameter, e.g. 0.6mm× 35 cm) with mild detergent and rinse extensively withwater. Dry all the items with a paper towel or tissue. Lubri-cate the spacers and the silicone rubber tubing with non-sili-cone grease. Apply the spacers along each of the two shortsides of the glass plate 2 mm away from the edges and 2 mmaway from the long side corresponding to the bottom of thegel. Begin to lay the silicone rubber tubing on the glass plateby using one spacer as a guide. Carefully twist the siliconerubber tubing at the bottom of the spacer and follow thelong side of the glass plate. While holding the silicone rubbertubing with one ˆnger along the long side again twist thetubing and lay it on the second short side of the glass plate,using the spacer as a guide. Place the second glass plate inperfect alignment and hold the mould together by hand pres-sure. Apply two clamps on each of the two short sides of themould. Carefully apply four clamps on the longer side of thegel mould, thus forming the bottom of the gel mould. Verifythat the silicone rubber tubing is running along the edge ofthe glass plates and has not been extruded while placing theclamps.2) Preparation of the gel

In a discontinuous buŠer SDS polyacrylamide gel, it isrecommended to pour the resolving gel, let the gel set, andthen pour the stacking gel, since the compositions of the twogels in acrylamide-bisacrylamide, buŠer and pH are diŠer-ent.Preparation of the resolving gel: In a conical ‰ask, preparethe appropriate volume of solution containing the desiredconcentration of acrylamide for the resolving gel, using thevalues given in Table 1. Mix the components in the ordershown. Where appropriate, before adding the ammoniumpersulfate solution and the tetramethylethylenediamine


(TEMED), ˆlter the solution if necessary under vacuumthrough a cellulose acetate membrane (pore size: 0.45 mm);keep the solution under vacuum by swirling the ˆltrationunit until no more bubbles are formed in the solution. Addappropriate amounts of ammonium persulfate solution andTEMED as indicated in Table 1, swirl and pour immediatelyinto the gap between the two glass plates of the mould.Leave su‹cient space for the stacking gel (the length of theteeth of the sample comb plus 1 cm). Using a pipette, care-fully overlay the solution with water-saturated isobutanol.Leave the gel in a vertical position at room temperature toallow polymerization to occur.Preparation of the stacking gel: After polymerization iscomplete (about 30 minutes), pour oŠ the isobutanol andwash the top of the gel several times with water to removethe isobutanol overlay and any unpolymerized acrylamide.Drain as much ‰uid as possible from the top of the gel, andthen remove any remaining water with the edge of a papertowel.

In a conical ‰ask, prepare the appropriate volume of solu-tion containing the desired concentration of acrylamide,using the values given in Table 2. Mix the components in theorder shown. Where appropriate, before adding the ammo-nium persulfate solution and the TEMED, ˆlter the solutionif necessary under vacuum through a cellulose acetate mem-brane (pore size: 0.45 mm); keep the solution under vacuumby swirling the ˆltration unit until no more bubbles areformed in the solution. Add appropriate amounts of ammo-nium persulfate solution and TEMED as indicated in Table2, swirl and pour immediately into the gap between the twoglass plates of the mould directly onto the surface of thepolymerized resolving gel. Immediately insert a clean samplecomb into the stacking gel solution, taking care to avoidtrapping air bubbles. Add more stacking gel solution to ˆllcompletely the spaces of the sample comb. Leave the gel in avertical position and allow to polymerize at room tempera-ture.3) Mounting the gel in the electrophoresis apparatus andelectrophoretic separation

After polymerization is complete (about 30 minutes),remove the sample comb carefully. Rinse the wells immedi-ately with water or with the running buŠer for SDS-Polyacrylamide Gel Electrophoresis to remove any un-polymerized acrylamide. If necessary, straighten the teeth ofthe sample comb of the stacking gel with a blunt hypodermicneedle attached to a syringe. Remove the clamps on oneshort side, carefully pull out the silicone rubber tubing andreplace the clamps. Proceed similarly on the other short side.Remove the silicone rubber tubing from the bottom part ofthe gel. Mount the gel in the electrophoresis apparatus. Addthe electrophoresis buŠers to the top and bottom reservoirs.Remove any bubbles that become trapped at the bottom ofthe gel between the glass plates. This is best done with a benthypodermic needle attached to a syringe. Never pre-run thegel before loading solutions, such as samples, since this willdestroy the discontinuity of the buŠer systems. Before load-ing solutions, such as samples, carefully rinse the stackinggel wells with the running buŠer for SDS-Polyacrylamide

Gel Electrophoresis. Prepare the test and reference solutionsin the recommended sample buŠer and treat as speciˆed inthe individual monograph. Apply the appropriate volume ofeach solution to the stacking gel wells. Start the electropho-resis using suitable operating conditions for the electropho-resis equipment to be used. There are commercially availablegels of diŠerent surface area and thickness that are appropri-ate for various types of electrophoresis equipment. Elec-trophoresis running time and currentWvoltage may need tobe altered depending on the type of apparatus used, in orderto achieve optimum separation. Check that the dye front ismoving into the resolving gel. When the dye is reaching thebottom of the gel, stop the electrophoresis. Remove the gelassembly from the apparatus and separate the glass plates.Remove the spacers, cut oŠ and discard the stacking gel andimmediately proceed with staining.

5. Detection of Proteins in GelsCoomassie staining is the most common protein staining

method, with a detection level of the order of 1 mg to 10 mgof protein per band. Silver staining is the most sensitivemethod for staining proteins in gels and a band containing l0ng to 100 ng can be detected. All of the steps in gel stainingare done at room temperature with gentle shaking in anyconvenient container. Gloves must be worn when staininggels, since ˆngerprints will stain.1) Coomassie staining

Immerse the gel in a large excess of Coomassie staining TSand allow to stand for at least 1 hour. Remove the stainingsolution.

Destain the gel with a large excess of destaining TS.Change the destaining solution several times, until thestained protein bands are clearly distinguishable on a clearbackground. The more thoroughly the gel is destained, thesmaller is the amount of protein that can be detected by themethod. Destaining can be speeded up by including 2 to 3 gof anion-exchange resin or a small sponge in the destainingTS.

NOTE: the acid-alcohol solutions used in this proceduredo not completely ˆx proteins in the gel. This can lead tolosses of some low-molecular-mass proteins during the stain-ing and destaining of the gel. Permanent ˆxation is obtaina-ble by allowing the gel to stand in trichloroacetic acid TS forˆxing for 1 hour before it is immersed in Coomassie stainingTS.2) Silver staining

Immerse the gel in a large excess of ˆxing TS and allow tostand for 1 hour. Remove the ˆxing solution, add fresh ˆx-ing solution and incubate either for at least 1 hour or over-night, if convenient. Discard the ˆxing solution and wash thegel in water for 1 hour. Soak the gel for 15 minutes in a 1 volz glutaraldehyde solution. Wash the gel twice for 15minutes in water. Soak the gel in fresh silver nitrate TS forsilver staining for 15 minutes, in darkness. Wash the gelthree times for 5 minutes in water. Immerse the gel for about1 minute in developer TS until satisfactory staining has beenobtained. Stop the development by incubation in blockingTS for 15 minutes. Rinse the gel with water.


6. Drying of Stained SDS-Polyacrylamide GelsDepending on the staining method used, gels are pretreat-

ed in a slightly diŠerent way. For Coomassie staining, afterthe destaining step, allow the gel to stand in a diluted solu-tion of concentrated glycerin (1 in 10) for at least 2 hours(overnight incubation is possible). For silver staining, add tothe ˆnal rinsing a step of 5 minutes in a diluted solution ofconcentrated glycerin (1 in 50).

Immerse two sheets of porous cellulose ˆlm in water andincubate for 5 to 10 minutes. Place one of the sheets on adrying frame. Carefully lift the gel and place it on the cellu-lose ˆlm. Remove any trapped air bubbles and pour 2 to 3mL of water around the edges of the gel. Place the secondsheet on top and remove any trapped air bubbles. Completethe assembly of the drying frame. Place in an oven or leaveat room temperature until dry.

7. Molecular-Mass DeterminationMolecular masses of proteins are determined by compari-

son of their mobilities with those of several marker proteinsof known molecular mass. Mixtures of proteins with pre-cisely known molecular masses blended for uniform stainingare commercially available for calibrating gels. They are ob-tainable in various molecular mass ranges. Concentratedstock solutions of proteins of known molecular mass arediluted in the appropriate sample buŠer and loaded on thesame gel as the protein sample to be studied.

Immediately after the gel has been run, the position of thebromophenol blue tracking dye is marked to identify theleading edge of the electrophoretic ion front. This can bedone by cutting notches in the edges of the gel or by insertinga needle soaked in India ink into the gel at the dye front.After staining, measure the migration distances of each pro-tein band (markers and unknowns) from the top of theresolving gel. Divide the migration distance of each proteinby the distance traveled by the tracking dye. The normalizedmigration distances so obtained are called the relative mobil-ities of the proteins (relative to the dye front) and conven-tionally denoted as Rf. Construct a plot of the logarithm ofthe relative molecular masses (Mr) of the protein standardsas a function of the Rf values. Note that the graphs areslightly sigmoid. Unknown molecular masses can be esti-mated by linear regression analysis or interpolation from thecurves of log Mr against Rf as long as the values obtained forthe unknown samples are positioned along the linear part ofthe graph.

8. Suitability of the Test (Validation)The test is not valid unless the front end of the molecular

mass marker migrates 80z of the migrating distance of thedye, and over the required separation range (e.g., the rangecovering the product and its dimer or the product and itsrelated impurities) the separation obtained for the relevantprotein bands shows a linear relationship between thelogarithm of the molecular mass and the Rf as described in7. Additional requirements with respect to the solutionunder test may be speciêd in individual monographs.

9. Quantiˆcation of ImpuritiesWhere the impurity limit is speciêd in the individual

monograph, a reference solution corresponding to that levelof impurity should be prepared by diluting the test solution.For example, where the limit is 5z, a reference solutionwould be a 1:20 dilution of the test solution. No impurity(any band other than the main band) in the electrophoreto-gram obtained with the test solution may be more intensethan the main band obtained with the reference solution.

Under validated conditions, impurities may be quantiêdby normalization to the main band, using an integrating den-sitometer. In this case, the responses must be validated forlinearity.

Test solutions:Coomassie staining TS Dissolve 125 mg of coomassie

brilliant blue R-250 in 100 mL of a mixture of water,methanol and acetic acid (100) (5:4:1), and ˆlter.

Developer TS Dissolve 2 g of citric acid monohydrate inwater to make 100 mL. To 2.5 mL of this solution add 0.27mL of formaldehyde solution and water to make 500 mL.

Fixing TS To 250 mL of methanol add 0.27 mL of for-maldehyde solution and water to make 500 mL.

Silver nitrate TS for silver staining To 40 mL of sodiumhydroxide TS add 3 mL of ammonia solution (28), then adddropwise 8 mL of a solution of silver nitrate (1 in 5) whilestirring, and add water to make 200 mL.

Destaining TS A mixture of water, methanol and aceticacid (100) (5:4:1).

Blocking TS To 10 mL of acetic acid (100) add water tomake 100 mL.

Trichloroacetic acid TS for ˆxing Dissolve 10 g oftrichloroacetic acid in a mixture of water and methanol (5:4)to make 100 mL.

1638

Table 1. Preparation of resolving gel

Solution componentsComponent volumes (mL) per gel mould volume of

5 mL 10 mL 15 mL 20 mL 25 mL 30 mL 40 mL 50 mL

6z AcrylamideWater 2.6 5.3 7.9 10.6 13.2 15.9 21.2 26.5Acrylamide solution(1) 1.0 2.0 3.0 4.0 5.0 6.0 8.0 10.01.5 molWL Tris solution (pH 8.8)(2) 1.3 2.5 3.8 5.0 6.3 7.5 10.0 12.5100 gWL SDS(3) 0.05 0.1 0.15 0.2 0.25 0.3 0.4 0.5100 gWL APS(4) 0.05 0.1 0.15 0.2 0.25 0.3 0.4 0.5TEMED(5) 0.004 0.008 0.012 0.016 0.02 0.024 0.032 0.04






(1) Acrylamide solution: 30z acrylamideWbisacrylamide (29:1) solution(2) 1.5 molWL Tris solution (pH 8.8): 1.5 molWL tris-hydrochloride buŠer solution, pH 8.8(3) 100 gWL SDS: 100 gWL solution of sodium dodecyl sulfate(4) 100 gWL APS: 100 gWL solution of ammonium persulfate. Ammonium persulfate provides the free radicals that drive

polymerization of acrylamide and bisacrylamide. Since ammonium persulfate solution decomposes slowly, fresh solu-tions must be prepared before use.

(5) TEMED: N,N,N?,N?-tetramethylethylenediamine


1639

Table 2. Preparation of stacking gel

Solution componentsComponent volumes (mL) per gel mould volume of

1 mL 2 mL 3 mL 4 mL 5 mL 6 mL 8 mL 10 mL

Water 0.68 1.4 2.1 2.7 3.4 4.1 5.5 6.8Acrylamide solution(1) 0.17 0.33 0.5 0.67 0.83 1.0 1.3 1.71.0 molWL Tris solution (pH 6.8)(2) 0.13 0.25 0.38 0.5 0.63 0.75 1.0 1.25100 gWL SDS(3) 0.01 0.02 0.03 0.04 0.05 0.06 0.08 0.1100 gWL APS(4) 0.01 0.02 0.03 0.04 0.05 0.06 0.08 0.1TEMED(5) 0.001 0.002 0.003 0.004 0.005 0.006 0.008 0.01

(1) Acrylamide solution: 30z acrylamideWbisacrylamide (29:1) solution(2) 1.0 molWL Tris solution (pH 6.8): 1 molWL tris-hydrochloride buŠer solution, pH 6.8(3) 100 gWL SDS: 100 gWL solution of sodium dodecyl sulfate(4) 100 gWL APS: 100 gWL solution of ammonium persulfate. Ammonium persulfate provides the free radicals that drive

polymerization of acrylamide and bisacrylamide. Since ammonium persulfate solution decomposes slowly, fresh solu-tions must be prepared before use.

(5) TEMED: N,N,N?,N?-tetramethylethylenediamine


16411641Supplement I, JP XIV Appendix

The Ministry of Health, Labour and WelfareThe Ministerial Notiˆcation on Partial Revision

of the Japanese Pharmacopoeia(No. 151, March 2002)

In accordance with the provision of Paragraph 1 ofArticle 41 of the Pharmaceutical AŠairs Law (LawNo. 145, 1960), we hereby revise a part of theJapanese Pharmacopoeia (Ministerial NotiˆcationNo. 111, March 2001) as follows and enforce the revi-sion on April 1, 2002. Although the revision shallcome into eŠect on April 1, 2002, in the case of thedrugs, which are prepared or imported before March31, 2003, the previous texts in the General Notices ofthe Part I or the Part II in the Japanese Phar-macopoeia may be applied. (The paragraphs on theGeneral Notices of the Part I are referred to theGeneral Notices of the Part II.)

March 29, 2002

Chikara SakaguchiThe Minister of Health, Labour and Welfare

Add the following paragraph to the General No-tices of the Part I :

39. In principle, unless otherwise speciˆed,animals used as a source of materials for preparingpharmaceutical preparations listed in the JapanesePharmacopoeia must be healthy.


Part I

Phenacetin

16431643

INDEX

Page citations refer to the pages of the Supplement I, and to pages ofthe JP XIV main volume, including those where the text being revised inthe Supplement originally appeared. This Supplement I commences withpage 1359 and succeeding Supplements will continue to be paged in se-quence.

1„1358 Main Volume of JP XIV1359„1668 Supplement I

A

AbsorbentCotton, 851Cotton, Puriêd, 851Cotton, Sterile, 852Cotton, Sterile, Puriêd, 852Gauze, 853Gauze, Sterile, 854

Absorptive Ointment, 855Acacia, 855, 1543

Powdered, 855, 1543Acebutolol Hydrochloride, 219Aceglutamide Aluminum, 1377Acetaminophen, 219Acetazolamide, 220Acetic Acid, 856

Glacial, 856Acetohexamide, 221Acetylcholine Chloride for

Injection, 222Acetylkitasamycin, 223, 1378Acetylleucomycin, 223Acetylsalicylic Acid, 252

Tablets, 252Acetylspiramycin, 224, 1378Achyranthes Root, 857Aclarubicin Hydrochloride, 224, 1379Acrinol, 224

and Zinc Oxide Oil, 857and Zinc Oxide Ointment, 858

Actinomycin D, 225, 1381Adhesive Plaster, 858Adrenaline, 447

Hydrochloride Injection, 447Hydrochloride Solution, 448

AdsorbedDiphtheria-Puriêd Pertussis-

Tetanus Combined Vaccine,1005

Diphtheria-Tetanus CombinedToxoid, 913

Diphtheria Toxoid for Adult Use,912

Habu-venom Toxoid, 938Hepatitis B Vaccine, 938Puriêd Pertussis Vaccine, 1005Tetanus Toxoid, 1066

A‰oqualone, 225Agar, 859

Powdered, 859Ajmaline, 226

Tablets, 227Akebia Stem, 859Albumin Tannate, 227Alcohol, 914

Benzyl, 874Dehydrated, 914for Disinfection, 916Isopropyl, 556Stearyl, 1059

Aldioxa, 228Alimemazine Tartrate, 229Alisma Rhizome, 860

Powdered, 860Allopurinol, 229Aloe, 860, 1544

Powdered, 861, 1544Alprazolam, 230Alpinia O‹cinarum Rhizome, 1544Alprenolol Hydrochloride, 231Alprostadil Alfadex, 231Alum, 862

Burnt, 863Solution, 861

AluminumAcetylsalicylate, 253Aspirin, 253Monostearate, 861Potassium Sulfate, 862Potassium Sulfate, Dried, 863Silicate, Natural, 233Silicate, Synthetic, 235Sucrose Sulfate Ester, 766

Amantadine Hydrochloride, 235

Ambenonium Chloride, 236Amidotrizoic Acid, 237Amikacin Sulfate, 238Aminoacetic Acid, 931Aminobenzylpenicillin, 246

Anhydrous, 246Sodium, 246

Aminophylline, 239Injection, 239

AmitriptylineHydrochloride, 240Hydrochloride Tablets, 240

Ammonia Water, 241Amobarbital, 242

Sodium for Injection, 242Amomum Seed, 863Amorphous Insulin Zinc Injection

(Aqueous Suspension), 542Amoxapine, 243Amoxicillin, 244, 1381Amphotericin B, 245Ampicillin, 246, 1382

Anhydrous, 246, 1384Ethoxycarbonyloxyethyl

Hydrochloride, 259Sodium, 246, 1385

AmpicillinphthalidylHydrochloride, 782

Amyl Nitrite, 246Anemarrhena Rhizome, 864Anesthamine, 465Anesthetic Ether, 462Angelica Dahurica Root, 864Anhydrous

Aminobenzylpenicillin, 246Ampicillin, 246CaŠeine, 294Citric Acid, 367, 1438Dibasic Calcium Phosphate, 879Lactose, 961

Antiformin, Dental, 864Antipyrine, 247Apricot

16441644 Supplement I, JP XIVIndex

Kernel, 864Kernel Water, 865

Arbekacin Sulfate, 247, 1386Areca, 866, 1545Arginine

Hydrochloride, 248Hydrochloride Injection, 249

L-ArginineHydrochloride, 248Hydrochloride Injection, 249

Aromatic Castor Oil, 889Arotinolol Hydrochloride, 249Arsenical Paste, 866Arsenic Trioxide, 250Arsenous Acid, 250Artemisia Capillaris Flower, 867Ascorbic Acid, 250

Injection, 251Powder, 251

Asiasarum Root, 867, 1545Asparagus Tuber, 1546Aspergillus Galactosidase, 921Aspirin, 252

Aluminum, 253Tablets, 252

Aspoxicillin, 254Astragalus Root, 867Astromicin Sulfate, 255, 1387Atractylodes

Lancea Rhizome, 869Rhizome, 868

AtropineSulfate, 255Sulfate Injection, 256

Azathioprine, 256Tablets, 256

Aztreonam, 258

B

Bacampicillin Hydrochloride, 259,1389

Bacitracin, 1390Baclofen, 260

Tablets, 260Bamethan Sulfate, 262Barbital 263Barium Sulfate, 264Bark

Cinnamon, 896Cinnamon, Powdered, 897Magnolia, 970Magnolia, Powdered, 971Mallotus, 971Moutan, 979Moutan, Powdered, 980Mulberry, 981Phellodendron, 1007Phellodendron, Powdered, 1008

Bearberry Leaf, 870Bear Bile, 869Beclometasone Dipropionate, 264Beef Tallow, 870

BeeswaxWhite, 871Yellow, 871

Bekanamycin Sulfate, 265, 1390Belladonna

Extract, 871Root, 872

Benoxinate Hydrochloride, 661Benserazide Hydrochloride, 265Bentonite, 873Benzalkonium

Chloride, 266Chloride Solution, 266Chloride Solution 50,

Concentrated, 267Benzbromarone, 268, 1391Benzethonium

Chloride, 269Chloride Solution, 269

Benzocaine, 465Benzoic Acid, 270Benzoin, 873Benzyl

Alcohol, 874Benzoate, 874

BenzylpenicillinBenzathine, 1392Potassium, 270, 1393

BerberineChloride, 271Tannate, 272

Betahistine Mesilate, 273Betamethasone, 274

Dipropionate, 275Sodium Phosphate, 276, 1395Valerate, 276, 1395

Bethanechol Chloride, 278Bifonazole, 278Biperiden Hydrochloride, 279Bisacodyl, 280

Suppositories, 280, 1396Bismuth

Subgallate, 281Subnitrate, 282

BitterCardamon, 875Orange Peel, 875Tincture, 875

BleomycinHydrochloride, 283, 1396Sulfate, 283, 1398

Blood, Whole Human, 1080Boric Acid, 284Botulism Antitoxin, Equine, Freeze-

dried, 876Bromazepam, 284Bromhexine Hydrochloride, 285Bromocriptine Mesilate, 286Bromovalerylurea, 287Bucumolol Hydrochloride, 287Bufetolol Hydrochloride, 288Bufexamac, 289

Cream, 289

Ointment, 289Bumetanide, 291Bunazosin Hydrochloride, 291Burdock Fruit, 1546Bupleurum Root, 876Bupranolol Hydrochloride, 292Burnt Alum, 863Busulfan, 293Butropium Bromide, 294Butyl Parahydroxybenzoate, 876Butyrate, Ribo‰avin, 734

C

Cacao Butter, 877CaŠeine, 294

and Sodium Benzoate, 296Anhydrous, 294

Calciferol, 449Calcium

Carbonate, Precipitated, 297Carboxymethylcellulose, 885Carmellose, 885Chloride, 298Chloride Injection, 298, 1399CMC, 885Folinate, 299Fosfomycin, 494Gluconate, 299Hydroxide, 877Lactate, 300Leucovorin, 299Oxide, 878Pantothenate, 301Para-aminosalicylate, 301Para-aminosalicylate Granules,

302Polystyrene Sulfonate, 303, 1399Stearate, 881

Calumba, 881Powdered, 881

Camellia Oil, 882Camostat Mesilate, 304Camphor, Synthetic, 306d-Camphor, 305, 1400dl-Camphor, 306, 1400Capsicum, 882, 1546

and Salicylic Acid Spirit, 884Powdered, 883, 1546Tincture, 883

Capsules, 885Cloˆbrate, 373Flurazepam, 488Indometacin, 535Sodium Iodide (123I), 755Sodium Iodide (131I), 755Sodium Iopodate, 757Vitamin A, 1075Vitamin A Oil, 1074

Captopril, 307Carbamazepine, 308Carbazochrome Sodium Sulfonate,

308, 1400

16451645Supplement I, JP XIV Index

Carbetapentane Citrate, 674Carbetaoebtene Citrete, 674Carbidopa, 309, 1401L-Carbocisteine, 310Carbolic Acid, 1010

for Disinfection, 1010Liqueˆed, 1011

Carbon Dioxide, 312Carboxymethylcellulose, 885

Calcium, 886Sodium, 887

Cardamon, 885Carmellose, 885


Carmofur, 313Carnauba Wax, 888Carteolol Hydrochloride, 313Carumonam Sodium, 314, 1401Cassia Seed, 888Castor Oil, 888Catalpa Fruit, 889Cefaclor, 314, 1403Cefadroxil, 314Cefalexin, 315Cefaloridine, 317, 1405Cefalotin Sodium, 317, 1406Cefamandole Sodium, 317, 1407Cefapirin Sodium, 317Cefatrizine Propylene Glycolate, 318Cefazolin

Sodium, 319Sodium Hydrate, 321

Cefbuperazone Sodium, 322, 1409Cefcapene Pivoxil Hydrochloride,

322Cefdinir, 324Cefditoren Pivoxil, 325Cefepime Dihydrochloride, 326Cefetamet Pivoxil Hydrochloride,

328Ceˆxime, 329Cefmenoxime Hydrochloride, 330,

1410Cefmetazole Sodium, 321Cefminox Sodium, 332Cefodizime Sodium, 1412Cefoperazone Sodium, 332Cefoselis Sulfate, 334Cefotaxime Sodium, 335, 1413Cefotetan, 335, 1415Cefotiam

Hexetil Hydrochloride, 335, 1417Hydrochloride, 335

Cefoxitin Sodium, 336, 1419Cefozopran Hydrochloride, 337Cefpiramide Sodium, 338, 1421Cefpirome Sulfate, 338Cefpodoxime Proxetil, 1422Cefradine, 339Cefroxadine, 340, 1424Cefsulodin Sodium, 340Ceftazidime, 342

Cefteram Pivoxil, 344, 1425Ceftibuten, 344, 1426Ceftizoxime Sodium, 345Ceftriaxone Sodium, 346, 1427Cefuroxime Axetil, 347, 1429Cefuroxime Sodium, 348Cellulose

Acetate Phthalate, 893Microcrystalline, 889Powdered, 892

Cetanol, 894Cetraxate Hydrochloride, 349, 1431Chloral Hydrate, 350Chloramphenicol, 351, 1432

Palmitate, 1432Sodium Succinate, 1434

Chlordiazepoxide, 351Powder, 351, 1434Tablets, 352, 1435

ChlorhexidineGluconate Solution, 353Hydrochloride, 354

Chlorinated Lime, 894Chlormadinone Acetate, 355, 1435Chlorobutanol, 894Chlorphenesin Carbamate, 356, 1436Chlorpheniramine

and Calcium Powder, 895Maleate, 357Maleate Injection, 357Maleate Powder, 358Maleate Tablets, 359

d-Chlorpheniramine Maleate, 359Chlorpromazine

Hydrochloride, 360Hydrochloride Injection, 361Hydrochloride Tablets, 361

Chlorpropamide, 362Tablets, 362

Cholecalciferol, 363Cholera Vaccine, 896Cholesterol, 896Chorionic

Gonadotrophin, 934Gonadotrophin for Injection, 936

Chrysanthemum Flower, 1547Ciclacillin, 364, 1437Ciclosporin, 364

A, 364Cimetidine, 365Cimicifuga Rhizome, 896Cinchocaine Hydrochloride, 404Cinnamon

Bark, 896, 1547Oil, 897Powdered, 1547

Citric Acid, 366, 1437Anhydrous, 367, 1438

Citrus Unshiu Peel, 898Clarithromycin, 367Clemastine Fumarate, 368Clematis Root, 1547Clindamycin

Hydrochloride, 1439Phosphate, 369, 1440

Clinoˆbrate, 369Clocapramine Hydrochloride, 370Clofedanol Hydrochloride, 371Cloˆbrate, 372

Capsules, 373Clomifene

Citrate, 373, 1441Citrate Tablets, 374

Clomipramine Hydrochloride, 375Clonazepam, 375Clonidine Hydrochloride, 376Cloperastine Hydrochloride, 377Clotiazepam, 378Clotrimazole, 379Clove, 898

Oil, 898Powdered, 898

Cloxacillin Sodium, 380, 1441Cloxazolam, 380CMC, 885


Cnidium Rhizome, 899Cocaine Hydrochloride, 381Coconut Oil, 899Codeine

Phosphate, 382Phosphate Powder, 1, 382, 1442Phosphate Powder, 10, 383, 1442Phosphate Tablets, 384, 1443

Cod Liver Oil, 900, 1548Coix Seed, 900Colchicine, 384Colistin

Sodium Methanesulfonate, 385Sulfate, 1444

Colophonium, 1029Compound

Acrinol and Zinc Oxide Oil, 857Diastase and Sodium Bicarbonate

Powder, 907Hycodenone Injection, 995Iodine Glycerin, 950Methyl Salicylate Spirit, 977Oxycodone and Atropine

Injection, 996Oxycodone Injection, 995Phellodendron Powder for

Cataplasm, 1009, 1556Rhubarb and Senna Powder, 1028Salicylic Acid Spirit, 1033Scopolia Extract and Diastase

Powder, 1037Scopolia Extract and Tannic Acid

Ointment, 1039, 1560Scopolia Extract and Tannic Acid

Suppositories, 1041, 1560Thianthol and Salicylic Acid

Solution, 1067Vitamin B Powder, 1075

Concentrated


Glycerin, 504Glycerol, 504

Condurango, 900Fluidextract, 901

Coptis Rhizome, 901, 1548Powdered, 1548

CornOil, 903Starch, 903

Cornus Fruit, 903Cortisone

Acetate, 386Corydalis Tuber, 904Cotton, Absorbent, 851Cream, Bufexamac, 289Creosote, 904Cresol, 905

Solution, 905Croconazole Hydrochloride, 387Crude Glycyrrhiza Extract, 934Crystalline

Insulin Zinc Injection (AqueousSuspension), 543

Penicillin G Potassium, 270Crystal Violet, 617Cyanamide, 387Cyanocobalamin, 388

Injection, 389Cyclopentolate Hydrochloride, 389Cyclophosphamide, 390Cycloserine, 390Cyperus Rhizome, 906Cyproheptadine Hydrochloride, 391Cytarabine, 392

D

Dantrolene Sodium, 392Daunorubicin Hydrochloride, 1445Deferoxamine Mesilate, 393Dehydrated

Alcohol, 915Ethanol, 915

Dehydrocholate Sodium Injection,396

Dehydrocholic Acid, 394Injection, 396Puriˆed, 394

DemethylchlortetracyclineHydrochloride, 1446

DentalAntiformin, 864Iodine Glycerin, 951Paraformaldehyde Paste, 1001Phenol with Camphor, 1011Sodium Hypochlorite Solution, 864Triozinc Paste, 1072

Dermatol, 281Deslanoside, 396

Injection, 397Dexamethasone, 397Dextran 906

40, 398

40 Injection, 39870, 400Sulfate Sodium Sulfur 5, 401Sulfate Sodium Sulfur 18, 402

Dextrin, 906Dextromethorphan Hydrobromide,

402Diagnostic Sodium Citrate Solution,

754Diastase, 907

and Sodium Bicarbonate Powder,907

Diazepam, 403Dibasic

Calcium Phosphate, 878Sodium Phosphate, 907

Dibekacin Sulfate, 404, 1447Dibucaine Hydrochloride, 404Dichlorphenamide, 406

Tablets, 407Diclofenac Sodium, 405Diclofenamide, 406, 1448

Tablets, 407Dicloxacillin Sodium, 408Diethylcarbamazine

Citrate, 408Citrate Tablets, 409, 1449

DiethylstilbestrolDiphosphate, 493Diphosphate Tablets, 494

Difenidol Hydrochloride, 409Digenea, 908Digitalis, 908

Powdered, 910Digitoxin, 410

Tablets, 411Digoxin, 414

Injection, 415Tablets, 416

DihydrocodeinePhosphate, 417Phosphate Powder, 1, 417, 1449Phosphate Powder, 10, 418, 1450

Dihydroergotamine Mesilate, 418,1450

Dihydroergotoxine Mesilate, 412Dihydroxyaluminum Allantoinate,

228Dilazep Hydrochloride, 419Diltiazem Hydrochloride, 420, 1451Dilute

Hydrochloric Acid, 515Iodine Tincture, 950

Diluted Opium Powder, 987Dimemorfan Phosphate, 421Dimenhydrinate, 422Dimenhydrinate Tablets, 423Dimercaprol, 423

Injection, 424Dimorpholamine, 424

Injection, 425Dinoprost, 425Dionin, 467

Dioscorea Rhizome, 911Diphenhydramine, 426

and Bromovalerylurea Powder,911

Hydrochloride, 427Phenol and Zinc Oxide Liniment,

912Tannate, 427

Diphenylhydantoin, 685Powder, 686Sodium for Injection, 686Tablets, 686

DiphtheriaAntitoxin, Equine, Freeze-dried,

912Toxoid, 912

Diphtheria-Tetanus CombinedToxoid, 912

Dipyridamole, 428, 1451Disodium

Edetate, 913Ethylenediaminetetraacetate, 913

Disopyramide, 429Distigmine

Bromide, 429Bromide Tablets, 430

Disulˆram, 430Dobutamine Hydrochloride, 431,

1452Dopamine

Hydrochloride, 432Hydrochloride Injection, 433, 1452

Dover's Powder, 993Doxapram Hydrochloride, 433Doxorubicin Hydrochloride, 434,

1453Doxycycline Hydrochloride, 434,

1454Dried

Aluminum Hydroxide Gel, 233Aluminum Hydroxide Gel Fine

Granules, 233Aluminum Potassium Sulfate, 863Sodium Carbonate, 1055Sodium Sulˆte, 1058Thyroid, 1069Yeast, 1083

DromostanolonePropionate, 435Propionate Injection, 436

Droperidol, 435, 1456Drostanolone

Propionate, 435, 1456Propionate Injection, 436, 1456

Dydrogesterone, 437Tablets, 438

E

Ecarazine Hydrochloride, 811Ecothiopate Iodide, 438Edrophonium

Chloride, 439


Chloride Injection, 440EDTA Sodium, 913Elcatonin, 440En‰urane, 443, 1456Enoxacin, 444Enviomycin Sulfate, 1457Ephedra Herb, 914Ephedrine

Hydrochloride, 444Hydrochloride Injection, 445Hydrochloride Powder, 10z, 446Hydrochloride Tablets, 446

Epinephrine, 447Hydrochloride Injection, 447Hydrochloride Solution, 448Injection, 447Solution, 448

Epirenamine, 447Hydrochloride Injection, 447Hydrochloride Solution, 448

Epirizole, 448Epirubicin Hydrochloride, 1458Ergocalciferol, 449, 1460Ergometrine

Maleate, 450Maleate Injection, 451Maleate Tablets, 451

Ergotamine Tartrate, 452Erythromycin, 453, 1460

Ethylsuccinate, 453Lactobionate, 1462Stearate, 454

Estazolam, 454Estradiol

Benzoate, 455, 1462Benzoate Injection, 456Benzoate Injection (Aqueous

Suspension), 456Estriol, 457, 1463

Injection (Aqueous Suspension),458

Tablets, 458Etacrynic Acid, 459

Tablets, 460Ethacridine Lactate, 224Ethambutol Hydrochloride, 460Ethanol, 914

Dehydrated, 915for Disinfection, 916

Ethenzamide, 461Ether, 462

Anesthetic, 462Guaiacol Glyceryl, 505Polyoxyethylene Lauryl Alcohol,

964Ethinylestradiol, 463

Tablets, 463Ethionamide, 464Ethosuximide, 465Ethoxybenzamide, 461Ethyl

Aminobenzoate, 465Cysteine Hydrochloride, 466

L-Cysteine Hydrochloride, 466Parahydroxybenzoate, 916

Ethylenediamine, 917Ethylmorphine Hydrochloride, 467Etilefrine

Hydrochloride, 468Hydrochloride Tablets, 468

Etizolam, 1463Eucalyptus Oil, 917Eucommia Bark, 1548Evodia Fruit, 918, 1548Exsiccated Gypsum, 938Extract

Belladonna, 871Crude Glycyrrhiza, 934Glycyrrhiza, 933Nux Vomica, 984

F

Famotidine, 469for Injection, 469, 1464Powder, 469Tablets, 469

Faropenem Sodium, 472Fenbufen, 473Fennel, 918

Oil, 919Powdered, 918

Fentanyl Citrate, 473Ferrous Sulfate, 474Fine Granules, Aluminum Hydroxide

Gel, Dried, 233Flavin Adenine Dinucleotide

Sodium, 475Flavoxate Hydrochloride, 476Floctafenine, 477, 1465Flomoxef Sodium, 478, 1465Flopropione, 478Flower, Artemisia Capillaris, 867Flucytosine, 478Fludiazepam, 479Flunitrazepam, 480Fluocinolone Acetonide, 481Fluocinonide, 482Fluorescein Sodium, 483Fluorometholone, 484Fluorouracil, 485Fluoxymesterone, 486Fluphenazine Enanthate, 487Flurazepam, 487

Capsules, 488Hydrochloride, 489

Flurbiprofen, 489, 1466Foeniculated Ammonia Spirit, 919Folic Acid, 490, 1466

Injection, 491Tablets, 491

Formalin, 919Water, 920

Formoterol Fumarate, 492Forsythia Fruit, 920Fosfestrol, 493

Tablets, 494Fosfomycin


Fradiomycin Sulfate, 497, 1467Freeze-dried

BCG Vaccine (for PercutaneousUse), 869

Botulism Antitoxin, Equine, 876Diphtheria Antitoxin, Equine, 912Habu Antivenom, Equine, 938Inactivated Tissue Culture Rabies

Vaccine, 1024Japanese Encephalitis Vaccine,

957Live Attenuated Measles Vaccine,

972Live Attenuated Mumps Vaccine,

981Live Attenuated Rubella Vaccine,

1029Mamushi Antivenom, Equine, 972Smallpox Vaccine, 1053Smallpox Vaccine Prepared in Cell

Culture, 1053Tetanus Antitoxin, Equine, 1066

Fritillaria Bulb, 1549Fructose, 497

Injection, 498Fruit

Catalpa, 889Cornus, 903Evodia, 918Forsythia, 920Gardenia, 922Gardenia, Powdered, 923Rose, 1029Rose, Powdered, 1029Schisandra, 1035Zanthoxylum, 1083Zanthoxylum, Powdered, 1084

Furosemide, 498Fursultiamine Hydrochloride, 499

G

Gabexate Mesilate, 500b-Galactosidase (Aspergillus), 921b-Galactosidase (Penicillium), 921Gallium (67Ga) Citrate Injection, 501Gambir, 920

Powdered, 920Gardenia Fruit, 922, 1549Gas Gangrene Antitoxin, Equine, 923Gastrodia Tuber, 1550Gauze, Absorbent, 853Gel, Aluminum Hydroxide, Dried,

233Gelatin, 923

Puriˆed, 924Gentamicin Sulfate, 502, 1468Gentian, 925

and Sodium Bicarbonate Powder,


926Japanese, 957Powdered, 925Powdered, Japanese, 957

Geranium Herb, 926Ginger, 927

Powdered, 927Ginseng, 927

Powdered, 929Glacial Acetic Acid, 856Glehnia Root, 930Glibenclamide, 502Glucose, 503

Injection, 503Glycerin, 504, 1469

and Potash Solution, 930Concentrated, 504, 1469

Glycerol, 504Concentrated, 504

Glyceryl Monostearate, 931Glycine, 931Glycyrrhiza, 932, 1550

Extract, 933Powdered, 933, 1550

Gramicidin, 1470Gonadotrophin

Chorionic, 934for Injection, Chorionic, 936for Injection, Serum, 937Serum, 936

Gramicidin 1470Granules

Calcium Para-aminosalicylate, 302Pas-calcium, 302

Griseofulvin, 505, 1471Guaiacol Glyceryl Ether, 505Guaifenesin, 505Guanabenz Acetate, 506Guanethidine Sulfate, 507Gypsum, 937

Exsiccated, 938

H

Habu Antivenom, Equine, Freeze-dried, 938

Haloperidol, 507Halothane, 508Haloxazolam, 509, 1472Hemp Fruit, 1551Heparin

Sodium, 510Sodium Injection, 511

HerbEphedra, 914Geranium, 926Houttuynia, 939Mentha, 974Perilla, 1004Plantago, 1014Powdered Geranium, 926Powdered Swertia, 1064Swertia, 1063

Homatropine Hydrobromide, 511Homochlorcyclizine Hydrochloride,

512, 1472Honey, 938Houttuynia Herb, 939Human Normal Immunoglobulin,

949Hycoato Injection, 996Hydralazine

Hydrochloride, 513Hydrochloride for Injection, 513Hydrochloride Powder, 513Hydrochloride Tablets, 514

Hydrochloric Acid, 514Dilute, 515Lemonade, 939

Hydrochlorothiazide, 515, 1473Hydrocortisone, 516

Acetate, 517, 1474and Diphenhydramine Ointment,

939Butyrate, 518Sodium Phosphate, 519, 1474Sodium Succinate, 520Succinate, 521

Hydrocotarnine Hydrochloride, 522Hydrogenated Oil, 940Hydrophilic

Ointment, 940Petrolatum, 1005

Hydrous Lanolin, 962Hydroxocobalamin Acetate, 523Hydroxypropylcellulose, 940

Low Substituted, 942Hydroxypropylmethylcellulose

Phthalate, 9472208, 9432906, 9442910, 945

HydroxyzineHydrochloride, 524Pamoate, 524

Hymecromone, 525

I

Ibuprofen, 526Ichthammol, 948Idarubicin Hydrochloride, 527, 1475Idoxuridine, 527

Ophthalmic Solution, 528, 1475Ifenprodil Tartrate, 529Imipenem, 530, 1476Imipramine


Immature Orange, 948Immunoglobulin, Human Normal,

949Imperata Rhizome, 949, 1551Indenolol Hydrochloride, 532Indigocarmine, 533

Injection, 533

Indium (111In) Chloride Injection, 534Indometacin, 534

Capsules, 535Suppositories, 536, 1477

In‰uenza HA Vaccine, 949Injection

Acetylcholine Chloride for, 222(Aqueous Suspension), Estradiol

Benzoate, 456(Aqueous Suspension), Estriol,

458(Aqueous Suspension), Insulin Zinc,

542(Aqueous Suspension), Insulin Zinc,

Amorphous, 542(Aqueous Suspension), Insulin Zinc,

Crystalline, 543(Aqueous Suspension), Insulin Zinc,

Protamine, 544(Aqueous Suspension), Isophane

Insulin, 541Adrenaline Hydrochloride, 447Aminophylline, 239Amobarbital Sodium for, 242Arginine Hydrochloride, 249L-Arginine Hydrochloride, 249Ascorbic Acid, 251Atropine Sulfate, 256Calcium Chloride, 298Chlorpheniramine Maleate, 357Chlorpromazine Hydrochloride,

361Cyanocobalamin, 389Dehydrocholate Sodium, 396Dehydrocholic Acid, 396Deslanoside, 397Dextran 40, 399Digoxin, 415Dimercaprol, 424Dimorpholamine, 425Diphenylhydantoin Sodium for,

686Dromostanolone Propionate, 436Drostanolone Propionate, 436Edrophonium Chloride, 440Ephedrine Hydrochloride, 445Epinephrine, 447Epinephrine Hydrochloride, 447Epirenamine Hydrochloride, 447Ergometrine Maleate, 451Estradiol Benzoate, 456Folic Acid, 491Fructose, 498Gallium (67Ga) Citrate, 501Glucose, 503Heparin Sodium, 511Human Serum Albumin, Iodinated

(131I), 546Hycoato, 996Hycodenone, Compound, 995Hydralazine Hydrochloride for,

513Indigocarmine, 533


Indium (111In) Chloride, 534Insulin, 539Isoniazid, 554Levallorphan Tartrate, 570Lidocaine, 575Lidocaine Hydrochloride, 575Magnesium Sulfate, 586D-Mannite, 588D-Mannitol, 588Meglumine Amidotrizoate, 594Meglumine Iotalamate, 595Meglumine Sodium

Amidotrizote, 596Meglumine Sodium Iodamide, 597Metenolone Enanthate, 608Morphine and Atropine, 978Morphine Hydrochloride, 630Neostigmine Methylsulfate, 638Nicotinic Acid, 645Noradrenaline Hydrochloride, 650Norepinephrine, 650Norepirenamine Hydrochloride,

650Operidine, 679Opium Alkaloids and Atropine,

990Opium Alkaloids and

Scopolamine, 991Opium Alkaloids Hydrochlorides,

989Oxycodone and Atropine,

Compound, 996Oxycodone, Compound, 995Oxytocin, 665Paoaverine Hydrochloride, 671Pethidine Hydrochloride, 679Phenolsulfonphthalein, 682Phenytoin Sodium for, 686Prednisolone Sodium Succinate for,

705Procainamide Hydrochloride, 709Procaine Hydrochloride, 710Progesterone, 714Protamine Sulfate, 719Pyridoxine Hydrochloride, 725Reserpine, 730Ribo‰avin Phosphate, 736Ribo‰avin Sodium Phosphate,

736Sodium Bicarbonate, 750Sodium Chloride, Isotonic, 752Sodium Chloride, 0.9, 752Sodium Chloride, 10, 752Sodium Chromate (51Cr), 752Sodium Iodohippurate (131I), 756Sodium Pertechnetate (99mTc), 758Sulfobromophthalein Sodium, 775Sulpyrine, 777Suxamethonium Chloride, 781Suxamethonium Chloride for, 781Testosterone Enanthate, 787Thallium (201Tl) Chloride, 791Thiamine Hydrochloride, 794

Thiamylal Sodium for, 796Thiopental Sodium for, 798Tubocurarine Chloride, 831Tubocurarine Hydrochloride, 831Vasopressin, 837Vinblastine Sulfate for, 841Vitamin B12, 389Vitamin B1 Hydrochloride, 794Vitamin B2 Phosphate Ester, 736Vitamin B6, 725Vitamin C, 251Weak Opium Alkaloids and

Scopolamine, 992Xylitol, 845

Insulin, 536Insulin Human (GeneticalRecombination), 537

Injection, 539Zinc Injection (Aqueous

Suspension), 542Zinc Injection (Aqueous

Suspension), Amorphous, 542Zinc Injection (Aqueous

Suspension), Crystalline, 543Zinc Protamine Injection (Aqueous

Suspension), 544Iodamide, 545Iodinated (131I) Human Serum

Albumin Injection, 546Iodine, 546

Glycerin, Compound, 950Glycerin, Dental, 951Tincture, 949Tincture, Dilute, 950Salicylic Acid and Phenol Spirit,

952Iopamidol, 547Iopanoic Acid, 548, 1478

Tablets, 548, 1478Iotalamic Acid, 549Iotroxic Acid, 550Ipecac, 953

Powdered, 954Syrup, 955

Ipratropium Bromide, 550Iproveratril Hydrochloride, 839Isepamicin Sulfate, 552L-Isoleucine, 553Isoniazid, 553

Injection, 554, 1478Tablets, 554, 1478

Isophane Insulin Injection (AqueousSuspension), 541

l-Isoprenaline Hydrochloride, 555Isopropanol, 556Isopropyl Alcohol, 556Isopropylantipyrine, 556l-Isoproterenol Hydrochloride, 555Isosorbide, 557

Dinitrate, 558Dinitrate Tablets, 558

IsotonicSalt Solution, 752

Sodium Chloride Injection, 752Sodium Chloride Solution, 752

J

JapaneseAngelica Root, 956Encephalitis Vaccine, 956Encephalitis Vaccine, Freeze-dried,

957Gentian, 957,Gentian, Powdered, 1551Valerian, 957

Josamycin, 559, 1479Propionate, 559, 1480

Jujube, 958Seed, 1551

K

Kainic Acid, 560and Santonin Powder, 958

Kallidinogenase, 560Kanamycin

Monosulfate, 1481Sulfate, 563, 1482

Kaolin, 959Kernel

Apricot, 864Peach, 1002Peach, Powdered, 1002

Ketamine Hydrochloride, 563Ketoprofen, 564Ketotifen Fumarate, 1483Kitasamycin, 565, 1484

Tartrate, 1485

L

Lactic Acid, 959Lactose, 960

Anhydrous, 961Lactulose, 566Lanatoside

C, 567C Tablets, 567

LanolinHydrous, 962Puriˆed, 963

Lard, 964Latamoxef Sodium, 569, 1486Lauromacrogol, 964Leaf

Bearberry, 870Senna, Powdered, 1046Sweet Hydrangea, Powdered, 1063Senna, 1044Sweet Hydrangea, 1063

Lenampicillin Hydrochloride, 1488L-Leucine, 569Leucomycin, 565Levallorphan

Tartrate, 570


Tartrate Injection, 570Levodopa, 571Levomepromazine Maleate, 572Levothyroxine

Sodium, 572Sodium Tablets, 573

Lidocaine, 574Hydrochloride Injection, 575Injection, 575

LightAnhydrous Silicic Acid, 1050Liquid Para‹n, 1001

LimeChlorinated, 894Quick, 878Slaked, 877

Lincomycin Hydrochloride, 576, 1490Liothyronine

Sodium, 576Sodium Tablets, 577, 1490

LiqueˆedCarbolic Acid, 1011Phenol, 1011

Liquid Para‹n, 1000Lithium Carbonate, 578Lithospermum Root, 964Live

Attenuated Measles Vaccine, Freeze-dried, 972

Attenuated Mumps Vaccine, Freeze-dried, 981

Attenuated Rubella Vaccine, Freeze-dried, 1029

Oral Poliomyelitis Vaccine, 1015Longgu, 965Loquat Leaf, 1552Lorazepam, 579Low Substituted

Hydroxypropylcellulose, 942Loxoprofen Sodium, 580Lysine Hydrochloride, 581Lysozyme Hydrochloride, 1491L-Lysine Hydrochloride, 581

M

MacrogolOintment, 968400, 9651500, 9664000, 9676000, 96720000, 968

MagnesiumCarbonate, 582Oxide, 583Silicate, 584Stearate, 969Sulfate, 585Sulfate Injection, 586, 1492Sulfate Mixture, 970

Magnolia Bark, 970, 1552Flower, 1552

Powdered, 1552Mallotus Bark, 971Maltose, 586Mamushi Antivenom, Equine, Freeze-

dried, 972D-Mannite, 587

Injection, 588D-Mannitol, 587

Injection, 588, 1492Maprotiline Hydrochloride, 588Meclofenoxate Hydrochloride, 589Mecobalamin, 590Medazepam, 591Medicinal

Carbon, 972Soap, 973

Mefenamic Acid, 592Mefruside, 592

Tablets, 593Meglumine, 974

Amidotrizoate Injection, 594Iotalamate Injection, 595Sodium Amidotrizoate Injection,

596Sodium Iodamide Injection, 597

Melphalan, 597Menatetrenone, 598, 1492Mentha Herb, 974

Oil, 975Water, 975

dl-Menthol, 975l-Menthol, 976Mepenzolate Bromide, 599Mepirizole, 448Mepitiostane, 600, 1492Mepivacaine


Mequitazine, 602Merbromin, 604

Solution, 605Mercaptopurine, 603Mercurochrome, 604

Solution, 605Meropenem Trihydrate, 605Mestranol, 606Metenolone

Acetate, 607Enanthate, 607Enanthate Injection, 608

Methamphetamine Hydrochloride,609

L-Methionine, 609Methotrexate, 610, 1493Methoxsalen, 611Methylbenactyzium Bromide, 611Methylcellulose, 978Methylchlorophenylisoxazolylpenicil-

lin Sodium, 380Methyldichlorophenylisoxazolylpenicil-

lin Sodium, 408Methyldopa, 612

Tablets, 613

dl-MethylephedrineHydrochloride, 613Hydrochloride Powder, 614Hydrochloride Powder, 10, 614

MethylergometrineMaleate, 615Maleate Tablets, 615

MethylParahydroxybenzoate, 976Salicylate, 618

Methylprednisolone, 616Methylrosanilinium Chloride, 617Methyltestosterone, 618

Tablets, 619Meticrane, 619, 1493Metildigoxin, 620Metoclopramide, 621Metronidazole, 622Metyrapone, 623Mexiletine Hydrochloride, 623Miconazole, 624

Nitrate, 625Microcrystalline Cellulose, 889Micronomicin Sulfate, 626, 1494Midecamycin, 626

Acetate, 627Migrenin, 627, 1495Minocycline Hydrochloride, 628,

1495Mitomycin C, 629, 1496Mixture, Magnesium Sulfate, 970Monobasic Calcium Phosphate, 880Monosodium

Trichloroethyl Phosphate, 823Trichloroethyl Phosphate Syrup,

824Morphine

and Atropine Injection, 978, 1553Hydrochloride, 629, 1497Hydrochloride Injection, 630, 1497Hydrochloride Tablets, 631, 1498

Moutan Bark, 979Mulberry Bark, 980, 1554Mupirocin Calcium Hydrate, 631

N

Nadolol, 633Nalidixic Acid, 634Naloxone Hydrochloride, 634Naphazoline

and Chlorpheniramine Solution,981

Hydrochloride, 635Nitrate, 636

Naproxen, 636Narcotine, 654

Hydrochloride, 655Natural Aluminum Silicate, 233Neomycin Sulfate, 497Neostigmine

Methylsulfate, 637Methylsulfate Injection, 638


Netilmicin Sulfate, 638Nicardipine Hydrochloride, 639

Hydrochloride Injection, 640Niceritrol, 641Nicomol, 642

Tablets, 643Nicotinamide, 643Nicotinic Acid, 644

Injection, 645Nifedipine, 645Nitrazepam, 646Nitrogen, 982Nitroglycerin Tablets, 647Nitrous Oxide, 648Noradrenaline, 649

Hydrochloride, 650Hydrochloride Injection,

Norepinephrine, 649, 1498Injection, 650, 1499

Norepirenamine, 649Hydrochloride Injection, 650

Norethisterone, 650Nor‰oxacin, 651Norgestrel, 652

and Ethinylestradiol Tablets, 652,1499

Nortriptyline Hydrochloride, 654Noscapine, 654

Hydrochloride, 655Notopterygium Rhizome, 1554Nuphar Rhizome, 983Nux

Vomica, 983Vomica Extract, 984Vomica Extract Powder, 984Vomica Tincture, 985

Nystatin, 656

O

O‰oxacin, 1500Oil

Acrinol and Zinc Oxide, 857Acrinol and Zinc Oxide,

Compound, 857Aromatic Castor,Camellia, 889Castor, 888Cinnamon, 897Clove, 898Coconut, 899Cod Liver, 900Corn, 903Eucalyptus, 917Fennel, 919Hydrogenated, 940Mentha, 975Olive, 986Orange, 994Peanut, 1002Rape Seed, 1024Sesame, 1048Soybean, 1059

Turpentine, 1073Vitamin A, 1074Zinc Oxide, 1085

OintmentAbsorptive, 855Acrinol and Zinc Oxide, 858Bufexamac, 290Hydrocortisone and

Diphenhydramine, 939Hydrophilic, 940Macrogol, 968Polyethylene Glycol, 968Scopolia Extract and Tannic Acid

Compound, 1039Simple, 1052Sulfur, Salicylic Acid and

Thianthol, 1062White, 1080Zinc Oxide, 1085

Olive Oil, 986Operidine, 678

Injection, 679Ophiopogon Tuber, 987Opium

Alkaloids and Atropine Injection,990

Alkaloids and ScopolamineInjection, 991

Alkaloids Hydrochlorides, 988,1555

Alkaloids HydrochloridesInjection, 989

Ipecac Powder, 993Powdered, 987Tincture, 988

OrangeImmature, 948Oil, 994Peel Syrup, 994Peel Tincture, 995

Orciprenaline Sulfate, 656Oriental Bezoar, 995Oxapium Iodide, 657Oxaprozin, 658Oxazolam, 658Oxetacaine, 659Oxethazaine, 659Oxprenolol Hydrochloride, 660Oxybuprocaine Hydrochloride, 661Oxycodone Hydrochloride, 662Oxydol, 662Oxygen, 663Oxymetholone, 664Oxytetracycline Hydrochloride, 665,

1501Oxytocin Injection, 665Oyster Shell, 997

P

Panax Rhizome, 998Powdered, 1555

Pancreatin, 999

Pancuronium Bromide, 667Panipenem, 667Pantethine, 669Papaverine


Paracetamol, 219Para‹n, 999

Light Liquid, 1001Liquid, 1000

Paraformaldehyde, 671Paste

Arsenical, 866Paraformaldehyde, Dental, 1001Triozinc, Dental, 1072

Pas-calcium, 301Granules, 302

Peach Kernel, 1002Peanut Oil, 1002Peel

Bitter Orange, 875Citrus Unshiu, 898

Penbutolol Sulfate, 672Penicillin

G Potassium, 270Pentazocine, 672Pentobarbital Calcium, 673Pentoxyverine Citrate, 674Peony Root, 1003, 1555

Powdered, 1556Peplomycin Sulfate, 675, 1503Perilla Herb, 1004Perphenazine, 675

Maleate, 677Maleate Tablets, 678Tablets, 676

PethidineHydrochloride, 678, 1505Hydrochloride Injection, 679

PetrolatumWhite, 1005Yellow, 1006

Petroleum Benzin, 1006Pharbitis Seed, 1007Phellodendron

Bark, 1007, 1556Albumin Tannate and Bismuth

Subnitrate Powder, 1009, 1556Powdered, 1556Powder for Cataplasm, Compound,

1556Phenacetin, 680Phenazone, 247Phenethicillin Potassium, 1505Phenobarbital, 681

Powder, 681Powder, 10, 681

Phenol, 1010and Zinc Oxide Liniment, 1011for Disinfection, 1010Liqueˆed, 1011with Camphor, Dental, 1011

Phenolated


Water, 1012Water for Disinfection, 1012

Phenolsulfonphthalein, 682Injection, 682

Phenovalin and Magnesium OxidePowder, 1012, 1557

L-Phenylalanine, 683Phenylbutazone, 684Phenylephrine Hydrochloride, 684Phenytoin, 685

Powder, 686Sodium for Injection, 686Tablets, 686

Phytomenadione, 687Phytonadione, 687, 1507Picrasma Wood, 1013Pilocarpine Hydrochloride, 687Pimaricin, 1507Pindolol, 688Pinellia Tuber, 1013Pipemidic Acid Trihydrate, 689, 1508Piperacillin Sodium, 690, 1508Piperazine

Adipate, 691Phosphate, 692Phosphate Tablets, 692

Pirarubicin, 1509Pirenoxine, 693Pivmecillinam Hydrochloride, 693,

1510Plantago

Herb, 1014Seed, 1014

Plaster, Adhesive, 858Salicylic Acid, Adhesive, 1032

PlatycodonFluidextract, 1014Root, 1014Root, Powdered, 1015

PolyethyleneGlycol Ointment, 968Glycol 400, 965Glycol 1500, 966Glycol 4000, 967Glycol 6000, 967Glycol 20000, 968

PolygalaRoot, 1015Root, Powdered, 1016

Polygonum Root, 1557Polymixin B Sulfate, 694, 1511Polyoxyethylene Lauryl Alcohol

Ether, 964Polyoxyl 40 Stearate, 1016Polyporus Sclerotium, 1016Polysorbate 80, 1017Polyvidone, 1020Polyvinylpyrrolidone, 1020

K25, 1020K30, 1020K90, 1020

Poria Sclerotium, 1017Potash Soap, 1018

PotassiumBenzylpenicillin, 270Bromide, 694Canrenoate, 694Carbonate, 1018Chloride, 695Clavulanate, 696, 1512Guaiacolsulfonate, 696Hydroxide, 1018Iodide, 697Penicillin G, 270Penicillin G, Crystalline, 270Permanganate, 698Sulfate, 1019

Potato Starch, 1019Povidone, 1020Povidone-Iodine, 698, 1513Powder

Ascorbic Acid, 251Chlordiazepoxide, 351Chlorpheniramine and Calcium,

895Chlorpheniramine Maleate, 358Codeine Phosphate, 1, 382Codeine Phosphate, 10, 383Diastase and Sodium Bicarbonate,

907Diastase and Sodium Bicarbonate,

Compound, 907Dihydrocodeine Phosphate, 1,

417Dihydrocodeine Phosphate, 10,

418Diphenhydramine and

Bromovalerylurea, 911Diphenylhydantoin, 686Dover's, 993Ephedrine Hydrochloride, 446Ephedrine Hydrochloride, 10,

446for Cataplasm, Phellodendron,

Compound, 1009Gentian and Sodium Bicarbonate,

926Hydralazine Hydrochloride, 513Kainic Acid and Santonin, 958dl-Methylephedrine

Hydrochloride, 614dl-Methylephedrine

Hydrochloride, 10, 614Nux Vomica Extract, 984Opium Ipecac, 993Opium, Diluted, 987Phellodendron, Albumin Tannate

and Bismuth Subnitrate, 1009Phenobarbital, 681Phenobarbital, 10, 681Phenovalin and Magnesium Oxide,

1012Phenytoin, 686Reserpine, 731Reserpine, 0.1, 731Rhubarb and Senna, Compound,

Ribo‰avin, 734Salicylated Alum, 1032Scopolia Extract, 1036Scopolia Extract and Carbon, 1037Scopolia Extract and Diastase,

Compound, 1037Scopolia Extract and Ethyl

Aminobenzoate, 1037Scopolia Extract, Papaverine and

Ethyl Aminobenzoate, 1038Swertia and Sodium Bicarbonate,

1065Thiamine Hydrochloride, 794Vitamin B, Compound,Vitamin B1 Hydrochloride, 794Vitamin B2, 734Vitamin C, 251Zinc Oxide Starch, 1086

PowderedAcacia, 855, 1543Agar, 859Alisma Rhizome, 860Aloe, 861, 1544Amomum Seed, 863Atractylodes Lancea Rhizome, 869Atractylodes Rhizome, 868Calumba, 881Capsicum, 883, 1546Cellulose, 892Cinnamon Bark, 897, 1547Clove, 898Cnidium Rhizome, 899Coix Seed, 900Coptis Rhizome, 902, 1548Cyperus Rhizome, 906Digitalis, 910Dioscorea Rhizome, 911Gambir, 920Gardenia Fruit, 923Gentian, 925Geranium Herb, 926Ginger, 927Ginseng, 929Glycyrrhiza, 933Ipecac, 954Japanese Angelica Root, 956Japanses Gentian, 957, 1551Japanese Valerian, 958Magnolia Bark, 971, 1552Moutan Bark, 980Opium, 987Oyster Shell, 998Panax Rhizome, 998, 1555Peach Kernel, 1002Peony Root, 1004, 1556Phellodendron Bark, 1008, 1556Picrasma Wood, 1013Platycodon Root, 1015Polygala Root, 1016Polyporus Sclerotium, 1016Poria Sclerotium, 1017Rhubarb, 1027, 1558Rose Fruit, 1029


Scutellaria Root, 1043, 1560Senega, 1044Senna Leaf, 1046, 1561Smilax Rhizome, 1053Sophora Root, 1058Sweet Hydrangea Leaf, 1063Swertia Herb, 1064, 1562Tragacanth, 1072Zanthoxylum Fruit, 1084

Pranoprofen, 699Prazepam, 700

Tablets, 701Precipitated Calcium Carbonate, 297Prednisolone, 701

Acetate, 703, 1513Sodium Succinate for Injection,

705, 1513Succinate, 704Tablets, 702

Primidone, 706, 1514Probenecid, 706

Tablets, 707Procainamide

Hydrochloride, 708Hydrochloride Injection, 709Hydrochloride Tablets, 709

ProcaineHydrochloride, 710Hydrochloride Injection, 710, 1514

Procarbazine Hydrochloride, 711,1515

Procaterol Hydrochloride, 712Prochlorperazine

Maleate, 713Maleate Tablets, 713

Progesterone, 714Injection, 714

Proglumide, 715Promethazine Hydrochloride, 716Propantheline Bromide, 716Propranolol Hydrochloride, 717Propylene

Glycol, 1022Propyl Parahydroxybenzoate, 1021Propylthiouracil, 718

Tablets, 718Propyphenazone, 556Prostaglandin

E1 a-Cyclodextrin ClathrateCompound, 231

F2a, 425Protamine

Sulfate, 719Sulfate Injection, 719

Prothionamide, 720Protirelin, 720

Tartrate, 722Prunella Spike, 1023Pueraria Root, 1023, 1557Puriˆed

Absorbent Cotton, 851Dehydrocholic Acid, 395Gelatin, 924

Lanolin, 963Shellac, 1049Water, 1079

Pyrantel Pamoate, 722Pyrazinamide, 723, 1515Pyridostigmine Bromide, 724Pyridoxine


Pyroxylin, 1023Pyrrolnitrin, 1516

Q

Quick Lime, 878Quinidine Sulfate, 725Quinine

Ethyl Carbonate, 726Hydrochloride, 727Sulfate, 728

R

Ranitidine Hydrochloride, 1517Rape Seed Oil, 1024Red Ginseng, 1024Rehmannia Root, 1025Reserpine, 729, 1518

Injection, 730Powder, 731Powder, 0.1, 731Tablets, 731

RetinolAcetate, 732, 1519Palmitate, 733, 1519

RhizomeAlisma, 860Alisma, Powdered, 860Anemarrhena, 864Atractylodes, 868Atractylodes Lancea, 869Atractylodes Lancea, Powdered,

869Atractylodes, Powdered, 868Cimicifuga, 896Cnidium, 899Cnidium, Powdered, 899Coptis, 901Coptis, Powdered, 902Cyperus, 906Cyperus, Powdered, 906Dioscorea, 911Dioscorea, Powdered, 911Imperate, 949Nuphar, 983Panax, 998Panax, Powdered, 998Scopolia, 1040

Rhubarb, 1026, 1558Powdered, 1027, 1558

Ribo‰avin, 733Butyrate, 734Phosphate, 735

Phosphate Injection, 736Powder, 734Sodium Phosphate, 735Sodium Phosphate Injection, 736

Ribostamycin Sulfate, 737, 1520Rice Starch, 1028Rifampicin, 737, 1521Ringer's Solution, 1028Rokitamycin, 737Root

Achyranthes, 857Angelica Dahurica, 864Asiasarum, 867Astragalus, 867Belladonna, 872Bupleurum, 876Glehnia, 930Japanese Angelica, 956Japanese Angelica, Powdered, 956Lithospermum, 964Peony, 1003Peony, Powdered, 1004Platycodon, 1014Polygala, 1015Pueraria, 1023Rehmannia, 1025Saposhnikovia, 1034Saussurea, 1034Scutellaria, 1042Scutellaria, Powdered, 1043Sophora, 1058Sophora, Powdered, 1058Trichosanthes, 1072

Rose Fruit, 1029Rosin, 1029Roxithromycin, 738, 1522

S

Saccharated Pepsin, 1030Saccharin Sodium, 1030SaŒower, 1031SaŠron, 1031Salazosulfapyridine, 739Salbutamol Sulfate, 740Salicylated Alum Powder, 1032Salicylic Acid, 741

Plaster, Adhesive, 1032Spirit, 1033

Santonin, 742Tablets, 742

Saponated Cresol Solution, 905Saposhnikovia Root, 1034Saussurea Root, 1034Schisandra Fruit, 1035Schizonepeta Spike, 1035Sclerotium

Polyporus, Powdered, 1016Poria, 1017Poria, Powdered, 1017

ScopolamineButylbromide, 743, 1522Hydrobromide, 744


ScopoliaExtract, 1035Extract and Carbon Powder, 1036Extract and Ethyl Aminobenzoate

Powder, 1037, 1559Extract and Tannic Acid

Ointment, Compound, 1560Suppositories, 1039Suppositories, Compound, 1560

Extract Powder, 1036Extract, Papaverine and Ethyl

Aminobenzoate Powder, 1038Rhizome, 1040

Scutellaria Root, 1042, 1560Powdered, 1560

Secretin, 744Seed

Amomum, 863Amomum, Powdered, 863Cassia, 888Coix, 900Coix, Powdered, 900Pharbitis, 1007Plantago, 1014

Senega, 1043Powdered, 1044Syrup, 1044

Senna Leaf, 1044, 1560Powdered, 1046, 1561

SerumGonadotrophin, 936Gonadotrophin for Injection, 937

Sesame Oil, 1048Shellac

Puriˆed, 1049White, 1049

ShellOyster, 997Oyster, Powdered, 998

Siccaine, 1523Silicic Acid, Light Anhydrous, 1050Silver

Nitrate, 746Nitrate Ophthalmic Solution, 1041Protein, 1051Protein Solution, 1051

Simˆbrate, 746, 1524Simple

Ointment, 1052Syrup, 1052

Sinomenium Stem, 1052, 1562Sisomicin Sulfate, 747, 1524Slaked Lime, 877Smilax

Rhizome, 1053Rhizome, Powdered, 1053

SoapMedicinal, 973Potash, 1018

SodiumAcetate, 1053Aminobenzylpenicillin, 246Ampicillin, 246

Aurothiomalate, 748Benzoate, 749Bicarbonate, 749Bicarbonate and Bitter Tincture

Mixture, 1054Bicarbonate Injection, 750, 1524Bisulˆte, 1054Borate, 750Bromide, 750Carbonate, 1055Carbonate, Dried,Carmellose, 887Cefalotin, 317Cefamandole, 317Cefapirin, 317Cefazolin, 319Cefbuperazone, 322Cefmetazole, 331Cefoperazone, 332Cefotaxime, 335Cefoxitin, 336Cefpiramide, 338Cefsulodin, 340Ceftizoxime, 345Cefuroxime, 348Chloride, 751, 1524Chloride Injection, 0.9, 752Chloride Injection, 10, 752, 1525Chromate (51Cr) Injection, 752Citrate, 753Citrate Injection for Transfusion,

753Cloxacillin, 380CMC, 887Cromoglicate, 754Dantrolene, 392Diclofenac, 405Dicloxacillin, 408EDTA, 913Flavin Adenine Dinucleotide, 475Fluorescein, 483Fosfomycin, 496Fusidate, 1525Heparin, 510Hydrogen Sulˆte, 1054Hydroxide, 1056Iodide, 755Iodide (123I) Capsules, 755Iodide (131I) Capsules, 755Iodide (131I) Solution, 756Iodohippurate (131I) Injection, 756Iopodate, 756, 1526Iopodate Capsules, 757, 1526Iotalamate Injection, 757Latamoxef, 569Lauryl Sulfate, 1056Levothyroxine, 572Liothyronine, 576Loxoprofen, 580Metabisulˆte, 1057Methylchlorophenylisoxazolyl-

penicillin, 380Methyldichlorophenylisoxazolyl-

penicillin, 408Pertechnetate (99mTc) Injection, 758Phosphate, Dibasic, 907Picosulfate, 758Piperacillin, 690Polystyrene Sulfonate, 759Prasterone Sulfate, 760Pyrosulˆte, 1057Saccharin, 1030Salicylate, 761Sulˆte, Dried, 1058Thiosulfate, 762Thiosulfate Injection, 762Valproate, 762

SolutionAdrenaline Hydrochloride, 448Alum, 861Benzalkonium Chloride, 267Benzethonium Chloride, 269Chlorhexidine Gluconate, 353Cresol, 905Cresol, Saponated, 905Epinephrine, 448Epinephrine Hydrochloride, 448Epirenamine Hydrochloride, 448Glycerin and Potash, 930Idoxuridine Ophthalmic, 528Merbromin, 605Mercurochrome, 605Naphazoline and

Chlorpheniramine, 981Ringer's, 1028Salt, Isotonic, 752Silver Nitrate Ophthalmic, 1041Silver Protein, 1051Sodium Chloride, Isotonic, 752Sodium Hypochlorite, Dental, 864Sodium Iodide (131I), 756D-Sorbitol, 764Thianthol and Salicylic Acid,

Compound, 1067Tolnaftate, 816Zinc Sulfate Ophthalmic, 108650, Benzalkonium Chloride,

Concentrated, 267Sophora Root, 1058Sorbitan Sesquioleate, 1058D-Sorbitol, 763

Solution, 764Soybean Oil, 1059Spectinomycin Hydrochloride, 1526Spike

Prunella, 1023Schizonepeta, 1035

SpiritCapsicum and Salicylic Acid, 884Foeniculated Ammonia, 919Iodine, Salicylic Acid and Phenol,

952Methyl Salicylate, Compound, 977Salicylic Acid, 1033Salicylic Acid, Compound, 1033

Spironolactone, 765


StarchCorn, 903Potato, 1019Rice, 1028Wheat, 1080

Stearic Acid, 1059Stearyl Alcohol, 1059Stem

Akebia, 859Sinomenium, 1052

SterileAbsorbent Cotton, 852Absorbent Gauze, 853Puriˆed Absorbent Cotton, 852Puriˆed Water, 1079

Streptomycin Sulfate, 766, 1527Sucralfate, 766Sucrose, 1060Sulbactam Sodium, 768Sulbenicillin Sodium, 769, 1528Sulfadiazine Silver, 769Sulfafurazole, 773Sulfamethizole, 770Sulfamethoxazole, 770Sulfamonomethoxine, 771Sulfasalazine, 739Sulˆnpyrazone, 772

Tablets, 773Sulˆsomezole, 770Sulˆsoxazole, 773Sulfobromophthalein

Sodium, 774Sodium Injection, 775

Sulfur, 775and Camphor Lotion, 1062Salicylic Acid and Thianthol

Ointment, 1062Sulpiride, 776Sulpyrine, 776

Injection, 777Sultamicillin Tosilate, 778Sultiame, 779Suppositorise

Bisacodyl, 280Indometacin, 536Scopolia Extract and Tannic Acid,

1039Scopolia Extract and Tannic Acid,

Compound, 1041Suxamethonium

Chloride, 780Chloride for Injection, 781Chloride Injection, 781

Sweet Hydrangea Leaf, 1063Swertia

and Sodium Bicarbonate Powder,1065, 1563

Herb, 1063, 1562Powdered, 1562

SyntheticAluminum Silicate, 235Camphor, 306

Syrup

Ipecac, 955Orange Peel, 994Senega, 1044Simple, 1052Trichloroethyl Phosphate,

Monosodium, 824Triclofos Sodium, 824

T

TabletsAcetylsalicylic Acid, 252Ajmaline, 227Amitriptyline Hydrochloride, 240Aspirin, 252Azathioprine, 258Baclofen, 260Chlordiazepoxide, 352Chlorpheniramine Maleate, 359Chlorpromazine Hydrochloride,

361Chlorpropamide, 362Clomifene Citrate, 374Codeine Phosphate, 384Dichlorphenamide, 407Diclofenamide, 407Diethylcarbamazine Citrate, 409Diethylstilbestrol Diphosphate,

494Digitoxin, 411Digoxin, 416Dimenhydrinate, 423Diphenylhydantoin, 686Distigmine Bromide, 430Dydrogesterone, 438Ephedrine Hydrochloride, 446Ergometrine Maleate, 451Estriol, 458Etacrynic Acid, 460Ethinylestradiol, 463Etilefrine Hydrochloride, 468Folic Acid, 491Fosfestrol, 494Hydralazine Hydrochloride, 514Imipramine Hydrochloride, 531Iopanoic Acid, 548Isoniazid, 554Isosorbide Dinitrate, 558Lanatoside C, 567Levothyroxine Sodium, 573Liothyronine Sodium, 576Methyldopa, 613Methylergometrine Maleate, 615Methyltestosterone, 619Morphine Hydrochloride, 631Nitroglycerin, 647Norgestrel and Ethinylestradiol,

652Perphenazine, 676Perphenazine Maleate, 678Phenytoin, 686Piperazine Phosphate, 682Prazepam, 700

Prednisolone, 702Probenecid, 707Procainamide Hydrochloride, 709Prochlorperazine Maleate, 713Propylthiouracil, 718Reserpine, 731Santonin, 742Sulˆnpyrazone, 773Thiamazole, 792Tipepidine Hibenzate, 806Tolbutamide, 814Trihexyphenidyl Hydrochloride,

825Trimethadione, 827Warfarin Potassium, 843

Talampicillin Hydrochloride, 782,1529

Talc, 1065Tannalbin, 227Tannic Acid, 782Tartaric Acid, 1066Tegafur, 782, 1530Teicoplanin, 783, 1530Terbutaline Sulfate, 786Testosterone

Enanthate, 787Enanthate Injection, 787Propionate, 788Propionate Injection, 788

Tetanus Antitoxin, Equine, Freeze-dried, 1066

Tetracaine Hydrochloride, 789Tetracycline Hydrochloride, 789Thallium (201Tl) Chloride Injection,

791Theophylline, 791, 1531Thiamazole, 791

Tablets, 792Thiamine

Hydrochloride, 793, 1532Hydrochloride Injection, 794Hydrochloride Powder, 794Nitrate, 795, 1532

ThiamylalSodium, 796Sodium for Injection, 796

Thianthol, 1067Thiopental

Sodium, 797, 1532Sodium for Injection, 798

Thioridazine Hydrochloride, 798Thiotepa, 799L-Threonine, 799Thrombin, 1068Thymol, 1069Thyroid, Dried, 1069Tiaramide Hydrochloride, 800Ticarcillin Sodium, 801Ticlopidine Hydrochloride, 802Timepidium Bromide, 803Tincture

Bitter, 875Capsicum, 883


Iodine, 949Nux Vomica, 985Opium, 988Orange Peel, 995

Tinidazole, 804Tipepidine

Hibenzate, 804, 1533Hibenzate Tablets, 806

Titanium Oxide, 1070Toad Venom, 1071, 1563Tobramycin, 807, 1533Tocopherol, 807, 1534

Acetate, 808, 1535Calcium Succinate, 809Nicotinate, 810

dl-a-Tocopherol, 807Acetate, 808Nicotinate, 810

Todralazine Hydrochloride, 811Toˆsopam, 812Tolazamide, 813Tolbutamide, 814

Tablets, 814Tolnaftate, 815

Solution, 816Tolperisone Hydrochloride, 816Toxoid

Diphtheria, 912Diphtheria-Tetanus, Combined,

913Diphtheria-Tetanus, Combined,

Adsorbed, 913for Adult Use, Diphtheria,

Adsorbed, 912Habu-venom, Adsorbed, 938Tetanus, Adsorbed, 1066

Tragacanth, 1072Powdered, 1072

Tranexamic Acid, 817Transfusion, Sodium Citrate Injection

for, 753Trapidil, 818Trepibutone, 819Tretoquinol Hydrochloride, 827Triamcinolone, 819, 1535

Acetonide, 820Triamterene, 821Trichlormethiazide, 822Trichloroethyl Phosphate,

Monosodium, 823Trichomycin, 823Trichosanthes Root, 1072Triclofos

Sodium, 823Sodium Syrup, 824

Trichomycin, 1536Trihexyphenidyl


Trimebutine Maleate, 1537Trimetazidine Hydrochloride, 826Trimethadione, 827

Tablets, 827

Trimetoquinol Hydrochloride, 827,1538Tropicamide, 828L-Tryptophan, 829Tuber

Corydalis, 904Ophiopogon, 987Pinellia, 1013

TubocurarineChloride, 830Chloride Injection, 831Hydrochloride, 830Hydrochloride Injection, 831

Tulobuterol Hydrochloride, 831Turpentine Oil, 1073

U

Ubidecarenone, 832Ulinastatin, 833Uncaria Thorn, 1563Urea, 835Urokinase, 1073Ursodesoxycholic Acid, 835Uva Ursi Fluidextract, 1074

V

VaccineCholera, 896Diphtheria-Puriêd Pertussis-

Tetanus Combined, Adsorbed,1005

Hepatitis B, Adsorbed, 938In‰uenza HA, 949Japanese Encephalitis, 956Japanese Encephalitis, Freeze-dried,

957Live Attenuated Measles, Freeze-d-

ried, 972Live Attenuated Mumps, Freeze-d-

ried, 981Live Attenuated Rubella, Freeze-d-

ried, 1029Live Oral Poliomyelitis, 1015Pertussis, Puriêd, Adsorbed, 1005Rabies, Inactivated Tissue Culture,

Freeze-dried, 1024Smallpox, Freeze-dried, 1053Smallpox, Freeze-dried, Prepared in

Cell Culture, 1053Weil's Disease and Akiyami

Combined, 1080(for Percutaneous Use), BCG,

Freeze-dried, 869L-Valine, 836Vancomycin Hydrochloride, 837,

1539Vasopressin Injection, 837Verapamil Hydrochloride, 839Vinblastine

Sulfate, 840Sulfate for Injection, 841

Vincristine Sulfate, 842Vitamin

A Acetate, 732A Capsules, 1075A Oil, 1074, 1564A Oil Capsules, 1075, 1565A Palmitate, 733B1 Hydrochloride, 793B1 Hydrochloride Injection, 794B1 Hydrochloride Powder, 794B1 Nitrate, 795B2, 733B2 Phosphate Ester, 735B2 Phosphate Ester Injection, 736B2 Powder, 734B6, 724B6 Injection, 725B12, 388B12 Injection, 389C, 250C Injection, 251C Powder, 251D2, 449D3, 363E, 807E Acetate, 808E Calcium Succinate, 809E Nicotinate, 810K1, 687

W

WarfarinPotassium, 843Potassium Tablets, 843

Water, 1076Ammonia, 241Apricot Kernel, 865for Disinfection, Phenolated, 1012for Injection, 1078Formalin, 920Mentha, 975Phenolated, 1012Puriêd, 1079Sterile, Puriêd, 1079

Wax, Carnauba, 888Weak Opium Alkaloids and

Scopolamine Injection, 992Weil's Disease and Akiyami CombinedVaccine, 1080Wheat Starch, 1080White

Beeswax, 871Ointment, 1080Petrolatum, 1005Shellac, 1049Soft Sugar, 1061

Whole Human Blood, 1080Wine, 1081Wood

Picrasma, 1013Picrasma, Powdered, 1013


X

Xylitol, 844Injection, 845, 1540

Y

Yeast, Dried, 1083Yellow

Beeswax, 871Petrolatum, 1006

Z

Zanthoxylum Fruit, 1083Zedoary, 1084Zinc

Chloride, 1084

Oxide, 845Oxide Oil, 1085Oxide Ointment, 1085Oxide Starch Powder, 1086Sulfate, 846Sulfate Ophthalmic Solution, 1086

Zinostatin Stimalamer, 846

16591659

INDEX IN JAPANESE

ア

亜鉛華デンプン 1086亜鉛華軟膏 1085アカメガシワ 971アクチノマイシン D 225, 1381アクリノール 224アクリノール・亜鉛華軟膏 858アクリノール・チンク油 857アザチオプリン 256アザチオプリン錠 257亜酸化窒素 648アジピン酸ピペラジン 691アジマリン 226アジマリン錠 227亜硝酸アミル 246アスコルビン酸 250アスコルビン酸散 251アスコルビン酸注射液 251アズトレオナム 258アスピリン 252アスピリンアルミニウム 253アスピリン錠 252アスポキシシリン 254アセグルタミドアルミニウム 1377アセタゾラミド 220アセチルキタサマイシン 222, 1378アセチルスピラマイシン 224, 1378アセトアミノフェン 219アセトヘキサミド 221アセンヤク 920アセンヤク末 920亜ヒ酸パスタ 866アフロクアロン 225アヘンアルカロイド・アトロピン注射

液 990アヘンアルカロイド・スコポラミン注

射液 991アヘン散 987アヘンチンキ 988アヘン・トコン散 993アヘン末 987アマチャ 1063アマチャ末 1063アミドトリゾ酸 237アミドトリゾ酸ナトリウムメグルミン

注射液 596アミドトリゾ酸メグルミン注射液

594アミノ安息香酸エチル 465アミノフィリン 239アミノフィリン注射液 239アムホテリシン B 245アモキサピン 243

アモキシシリン 244, 1381アモバルビタール 242アラビアゴム 855, 1543アラビアゴム末 855, 1543亜硫酸水素ナトリウム 1054アルジオキサ 228アルプラゾラム 230アルプロスタジルアルファデクス

231アロエ 860, 1544アロエ末 861, 1544アロプリノール 228アンソッコウ 873安息香酸 270安息香酸エストラジオール 455,

1462安息香酸エストラジオール水性懸濁注

射液 456安息香酸エストラジオール注射液

456安息香酸ナトリウム 749安息香酸ナトリウムカフェイン 296安息香酸ベンジル 874アンチピリン 247アンピシリン 246, 1382アンピシリンナトリウム 246, 1385アンモニア・ウイキョウ精 919アンモニア水 241

イ

イオウ 775イオウ・カンフルローション 1062イオウ・サリチル酸・チアントール軟膏

1062イオタラム酸 549イオタラム酸ナトリウム注射液 757イオタラム酸メグルミン注射液 595イオトロクス酸 550イオパノ酸 548, 1478イオパノ酸錠 548, 1478イオパミドール 547イオポダートナトリウム 756, 1526イオポダートナトリウムカプセル

757, 1526イクタモール 948イソソルビド 557イソニアジド 553イソニアジド錠 554, 1478イソニアジド注射液 554, 1478イソフェンインスリン水性懸濁注射液

541イソプロパノール 556イソプロピルアンチピリン 556イドクスウリジン 527

イドクスウリジン点眼液 528, 1475イブプロフェン 526イミペネム 530, 1476イレイセン 1547インジゴカルミン 533インジゴカルミン注射液 533インスリン 536インスリン亜鉛水性懸濁注射液 542インスリン注射液 539インチンコウ 867インドメタシン 534インドメタシンカプセル 535インドメタシン坐剤 536, 1477インフルエンザ HAワクチン 949

ウ

ウイキョウ 918ウイキョウ末 918ウイキョウ油 918ウリナスタチン 833ウルソデオキシコール酸 835ウロキナーゼ 1073ウワウルシ 870ウワウルシ流エキス 1074

エ

エイジツ 1029エイジツ末 1029液状フェノール 1011エスタゾラム 454エストリオール 457, 1463エストリオール錠 458エストリオール水性懸濁注射液 458エタクリン酸 459エタクリン酸錠 460エタノール 814エチオナミド 464エチゾラム 1463エチニルエストラジオール 463エチニルエストラジオール錠 463エチルコハク酸エリスロマイシン

453エチル炭酸キニーネ 727エチレンジアミン 917エデト酸ナトリウム 913エーテル 462エテンザミド 461エトスクシミド 465エナント酸テストステロン 787エナント酸テストステロン注射液

787エナント酸フルフェナジン 487エナント酸メテノロン 607

16601660 Supplement I, JP XIVIndex in Japanese

エナント酸メテノロン注射液 608エノキサシン 444エピネフリン 447エピネフリン液 447エピネフリン注射液 447エピリゾール 448エリスロマイシン 453, 1460L-イソロイシン 553エルカトニン 440L-カルボシステイン 310エルゴカルシフェロール 449, 1460L-トリプトファン 829L-トレオニン 799L-バリン 836L-フェニルアラニン 683L-メチオニン 609L-ロイシン 569塩化亜鉛 1084塩化アンベノニウム 236塩化インジウム (111In)注射液 534塩化エドロホニウム 439塩化エドロホニウム注射液 440塩化カリウム 695塩化カルシウム 298塩化カルシウム注射液 298, 1399塩化スキサメトニウム 781塩化スキサメトニウム注射液 781塩化タリウム (201Tl)注射液 791塩化ツボクラリン 830塩化ツボクラリン注射液 831塩化ナトリウム 751, 152410塩化ナトリウム注射液 752,

1525塩化ベタネコール 278塩化ベルベリン 271塩化ベンザルコニウム 266塩化ベンザルコニウム液 267塩化ベンゼトニウム 269塩化ベンゼトニウム液 269塩化メチルロザニリン 617塩化リゾチーム 1491エンゴサク 904塩酸 514塩酸アクラルビシン 224, 1379塩酸アセブトロール 219塩酸アヘンアルカロイド 988, 1555塩酸アヘンアルカロイド注射液 989塩酸アマンタジン 235塩酸アミトリプチリン 240塩酸アミトリプチリン錠 240塩酸アルプレノロール 231塩酸アロチノロール 249l-塩酸イソプレナリン 555塩酸イダルビシン 527, 1475塩酸イミプラミン 530塩酸イミプラミン錠 531塩酸インデノロール 532塩酸エタンブトール 460塩酸エチルモルヒネ 467塩酸エチレフリン 468塩酸エチレフリン錠 468塩酸エピルビシン 1458塩酸エフェドリン 444

塩酸エフェドリン散 10 446塩酸エフェドリン錠 446塩酸エフェドリン注射液 445塩酸 L-アルギニン 248塩酸 L-アルギニン注射液 248塩酸 L-エチルシステイン 466塩酸 L-リジン 581塩酸オキシコドン 662塩酸オキシテトラサイクリン 665,

1501塩酸オキシブプロカイン 661塩酸オクスプレノロール 660塩酸カルテオロール 313塩酸キニーネ 727塩酸クリンダマイシン 1439塩酸クロカプラミン 370塩酸クロコナゾール 387塩酸クロニジン 376塩酸クロフェダノール 371塩酸クロペラスチン 377塩酸クロミプラミン 375塩酸クロルプロマジン 360塩酸クロルプロマジン錠 361塩酸クロルプロマジン注射液 361塩酸クロルヘキシジン 354塩酸ケタミン 563塩酸コカイン 381塩酸シクロペントラート 389塩酸ジフェニドール 409塩酸ジフェンヒドラミン 427塩酸ジブカイン 404塩酸シプロヘプタジン 391塩酸ジラゼプ 419塩酸ジルチアゼム 420, 1451塩酸スペクチノマイシン 1526塩酸セトラキサート 349, 1431塩酸セフェタメトピボキシル 328塩酸セフェピム 326塩酸セフォゾプラン 337塩酸セフォチアム 335塩酸セフォチアムヘキセチル 335,

1417塩酸セフカペンピボキシル 322塩酸セフメノキシム 330, 1410塩酸ダウノルビシン 1445塩酸タランピシリン 782, 1529塩酸チアミン 793, 1532塩酸チアミン散 794塩酸チアミン注射液 794塩酸チアラミド 800塩酸チオリダジン 798塩酸チクロピジン 802塩酸ツロブテロール 831塩酸テトラカイン 789塩酸テトラサイクリン 789塩酸デメチルクロルテトラサイクリン

1446塩酸ドキサプラム 433塩酸ドキシサイクリン 434, 1454塩酸ドキソルビシン 434, 1453塩酸トドララジン 811塩酸ドパミン 432塩酸ドパミン注射液 433, 1452

塩酸ドブタミン 431, 1452塩酸トリヘキシフェニジル 824塩酸トリヘキシフェニジル錠 825塩酸トリメタジジン 826塩酸トリメトキノール 827, 1538塩酸トルペリゾン 816塩酸ナファゾリン 635塩酸ナロキソン 634塩酸ニカルジピン 639塩酸ニカルジピン注射液 640塩酸ノスカピン 655塩酸ノルトリプチリン 654塩酸バカンピシリン 259, 1389塩酸パパベリン 670塩酸パパベリン注射液 671塩酸バンコマイシン 837, 1539塩酸ヒドララジン 513塩酸ヒドララジン散 513塩酸ヒドララジン錠 514塩酸ヒドロキシジン 524塩酸ヒドロコタルニン 522塩酸ピブメシリナム 693, 1510塩酸ビペリデン 279塩酸ピリドキシン 724塩酸ピリドキシン注射液 725塩酸ピロカルピン 687塩酸フェニレフリン 684塩酸ブクモロール 287塩酸ブナゾシン 291塩酸ブフェトロール 288塩酸ブプラノロール 292塩酸フラボキサート 476塩酸フルスルチアミン 499塩酸フルラゼパム 489塩酸ブレオマイシン 283, 1396塩酸プロカイン 710塩酸プロカインアミド 708塩酸プロカインアミド錠 709塩酸プロカインアミド注射液 709塩酸プロカイン注射液 710, 1514塩酸プロカテロール 712塩酸プロカルバジン 711, 1515塩酸プロプラノロール 717塩酸ブロムヘキシン 285塩酸プロメタジン 716塩酸ペチジン 678, 1505塩酸ペチジン注射液 679塩酸ベラパミル 839塩酸ベンセラジド 265塩酸ホモクロルシクリジン 512,

1472塩酸マプロチリン 588塩酸ミノサイクリン 628, 1495塩酸メキシレチン 623塩酸メクロフェノキサート 589塩酸メタンフェタミン 609dl-塩酸メチルエフェドリン 613dl-塩酸メチルエフェドリン散 10

614塩酸メピバカイン 601塩酸メピバカイン注射液 602塩酸モルヒネ 629, 1497塩酸モルヒネ錠 631, 1498

16611661Supplement I, JP XIV Index in Japanese

塩酸モルヒネ注射液 630, 1497塩酸ラニチジン 1517塩酸リモナーデ 939塩酸リンコマイシン 576, 1490塩酸レナンピシリン 1488エンフルラン 443, 1456

オ

オウギ 867オウゴン 1042, 1560オウゴン末 1043, 1560黄色ワセリン 1006オウバク 1007, 1556オウバク・タンナルビン・ビスマス散

1009, 1556オウバク末 1008, 1556オウレン 901, 1548オウレン末 902, 1548オキサゾラム 658オキサプロジン 658オキシトシン注射液 665オキシドール 662オキシメトロン 664オキセサゼイン 659オフロキサシン 1500オリブ油 986オレンジ油 994オンジ 1015オンジ末 1016

カ

カイニン酸 560カイニン酸・サントニン散 958カオリン 959カカオ脂 877加香ヒマシ油 889カゴソウ 1023カシュウ 1557ガジュツ 1084加水ラノリン 962ガスえそウマ抗毒素 923ガーゼ 853カッコン 1023, 1557過テクネチウム酸ナトリウム (99mTc)

注射液 758果糖 497果糖注射液 498カノコソウ 957カノコソウ末 958カフェイン 294カプセル 885カプトプリル 307過マンガン酸カリウム 698b-ガラクトシダーゼ（アスペルギルス）

921b-ガラクトシダーゼ（ペニシリウム）

921カリジノゲナーゼ 560カリ石ケン 1018カルナウバロウ 888カルバゾクロムスルホン酸ナトリウム

308, 1400カルバマゼピン 308カルバミン酸クロルフェネシン 356,

1436カルビドパ 309, 1401カルメロース 885カルメロースカルシウム 886カルメロースナトリウム 887カルモナムナトリウム 314, 1401カルモフール 313カロコン 1072カンゾウ 932, 1550カンゾウ末 1550乾燥亜硫酸ナトリウム 1058カンゾウエキス 933乾燥甲状腺 1069乾燥酵母 1083乾燥細胞培養痘そうワクチン 1053乾燥ジフテリアウマ抗毒素 912乾燥弱毒生おたふくかぜワクチン

981乾燥弱毒生風しんワクチン 1029乾燥弱毒生麻しんワクチン 972乾燥水酸化アルミニウムゲル 233乾燥水酸化アルミニウムゲル細粒

233カンゾウ粗エキス 934乾燥組織培養不活化狂犬病ワクチン

1024乾燥炭酸ナトリウム 1055乾燥痘そうワクチン 1053乾燥日本脳炎ワクチン 957乾燥破傷風ウマ抗毒素 1066乾燥はぶウマ抗毒素 938乾燥 BCGワクチン 869乾燥ボツリヌスウマ抗毒素 876カンゾウ末 933乾燥まむしウマ抗毒素 972乾燥硫酸アルミニウムカリウム 863カンテン 859カンテン末 859含糖ペプシン 1030d-カンフル 305, 1400dl-カンフル 306, 1400肝油 900, 1548カンレノ酸カリウム 694

キ

希塩酸 515キキョウ 1014キキョウ末 1015キキョウ流エキス 1014キクカ 1547キササゲ 889キジツ 948キシリトール 844キシリトール注射液 845, 1540吉草酸ベタメタゾン 277, 1395キタサマイシン 565, 1484牛脂 870吸水軟膏 855キョウカツ 1554

キョウニン 864キョウニン水 865希ヨードチンキ 950金チオリンゴ酸ナトリウム 748

ク

グアイフェネシン 505グアヤコールスルホン酸カリウム

696クエン酸 366, 1437クエン酸ガリウム (67Ga)注射液 501クエン酸クロミフェン 373, 1441クエン酸クロミフェン錠 374クエン酸ジエチルカルバマジン 408クエン酸ジエチルカルバマジン錠

409, 1449クエン酸ナトリウム 753クエン酸フェンタニル 473クエン酸ペントキシベリン 674クジン 1058クジン末 1058苦味重曹水 1054苦味チンキ 875クラブラン酸カリウム 696, 1512グラミシジン 1470クラリスロマイシン 367グリシン 931グリセオフルビン 505, 1471グリセリン 504, 1469グリセリンカリ液 930クリノフィブラート 369グリベンクラミド 502グルコン酸カルシウム 299グルコン酸クロルヘキシジン液 353クレオソート 904クレゾール 905クレゾール水 905クレゾール石ケン液 905クロキサシリンナトリウム 380,

1441クロキサゾラム 380クロチアゼパム 378クロトリマゾール 379クロナゼパム 375クロフィブラート 372クロフィブラートカプセル 373クロム酸ナトリウム (51Cr)注射液

752クロモグリク酸ナトリウム 754クロラムフェニコール 351, 1432クロルジアゼポキシド 351クロルジアゼポキシド散 351, 1434クロルジアゼポキシド錠 352, 1435クロルフェニラミン・カルシウム散

895クロルプロパミド 362クロルプロパミド錠 362クロロブタノール 894

ケ

ケイガイ 1035


経口生ポリオワクチン 1015ケイ酸マグネシウム 584軽質無水ケイ酸 1050軽質流動パラフィン 1001ケイヒ 896, 1547ケイヒ末 897, 1547ケイヒ油 897結晶性インスリン亜鉛水性懸濁注射液

543結晶セルロース 889血清性性腺刺激ホルモン 936ケツメイシ 888ケトプロフェン 564ケンゴシ 1007ゲンチアナ 925ゲンチアナ・重曹散 926ゲンチアナ末 925ゲンノショウコ 926ゲンノショウコ末 926

コ

コウカ 1031硬化油 940コウジン 1024合成ケイ酸アルミニウム 235コウブシ 906コウブシ末 906コウボク 970, 1552コウボク末 970, 1552ゴオウ 995ゴシツ 857ゴシュユ 918, 1548コハク酸クロラムフェニコールナトリ

ウム 1434コハク酸トコフェロールカルシウム

809コハク酸ヒドロコルチゾン 521コハク酸ヒドロコルチゾンナトリウム

520コハク酸プレドニゾロン 704ゴボウシ 1546ゴマ油 1048ゴミシ 1035コムギデンプン 1080コメデンプン 1028コリスチンメタンスルホン酸ナトリウ

ム 385コルヒチン 384コレカルシフェロール 363コレステロール 896コレラワクチン 896コロンボ 881コロンボ末 881コンズランゴ 900コンズランゴ流エキス 901

サ

サイクロセリン 390サイコ 876サイシン 867, 1545酢酸 856

酢酸グアナベンズ 506酢酸クロルマジノン 355, 1435酢酸コルチゾン 386酢酸トコフェロール 808, 1535酢酸ナトリウム 1053酢酸ヒドロキソコバラミン 523酢酸ヒドロコルチゾン 517, 1474酢酸フタル酸セルロース 893酢酸プレドニゾロン 703, 1513酢酸ミデカマイシン 627酢酸メテノロン 607酢酸レチノール 732, 1519サッカリンナトリウム 1030サフラン 1031サラシ粉 894サラシミツロウ 871サラゾスルファピリジン 739サリチル・ミョウバン散 1032サリチル酸 741サリチル酸精 1033サリチル酸ナトリウム 761サリチル酸絆創膏 1032サリチル酸メチル 618酸化亜鉛 845酸化カルシウム 878酸化チタン 1070酸化マグネシウム 583サンキライ 1053サンキライ末 1053三酸化ヒ素 250サンシシ 922, 1549サンシシ末 923サンシュユ 903サンショウ 1083サンショウ末 1084酸素 663サントニン 742サントニン錠 742サンヤク 911サンヤク末 911

シ

ジアスターゼ 907ジアスターゼ・重曹散 907ジアゼパム 403シアナミド 387シアノコバラミン 388シアノコバラミン注射液 389ジオウ 1025歯科用アンチホルミン 864歯科用トリオジンクパスタ 1072歯科用パラホルムパスタ 1001歯科用フェノール・カンフル 1011歯科用ヨード・グリセリン 951ジギタリス 908ジギタリス末 910ジギトキシン 410ジギトキシン錠 411シクラシリン 364, 1437ジクロキサシリンナトリウム 408シクロスポリン 364ジクロフェナクナトリウム 405

ジクロフェナミド 406, 1448ジクロフェナミド錠 407シクロホスファミド 390ジゴキシン 414ジゴキシン錠 416ジゴキシン注射液 415シコン 964次硝酸ビスマス 282ジスルフィラム 430ジソピラミド 429シタラビン 392シッカニン 1523ジドロゲステロン 437ジドロゲステロン錠 438ジノスタチンスチマラマー 846ジノプロスト 425ジピリダモール 428, 1451ジフェンヒドラミン 426ジフェンヒドラミン・フェノール・亜鉛

華リニメント 912ジフェンヒドラミン・ワレリル尿素散

911ジフテリアトキソイド 912ジフテリア破傷風混合トキソイド

913ジプロピオン酸ベタメタゾン 275シメチジン 365ジメルカプロール 423ジメルカプロール注射液 424ジメンヒドリナート 422ジメンヒドリナート錠 423次没食子酸ビスマス 281ジモルホラミン 424ジモルホラミン注射液 425弱アヘンアルカロイド・スコポラミン

注射液 992シャクヤク 1003, 1555シャクヤク末 1004, 1556シャゼンシ 1014シャゼンソウ 1014臭化イプラトロピウム 550臭化カリウム 694臭化ジスチグミン 429臭化ジスチグミン錠 430臭化水素酸スコポラミン 744臭化水素酸デキストロメトルファン

402臭化水素酸ホマトロピン 511臭化チメピジウム 803臭化ナトリウム 751臭化パンクロニウム 667臭化ピリドスチグミン 724臭化ブチルスコポラミン 743, 1522臭化ブトロピウム 294臭化プロパンテリン 716臭化メチルベナクチジウム 611臭化メペンゾラート 599ジュウヤク 939シュクシャ 863シュクシャ末 863酒石酸 1066酒石酸アリメマジン 229酒石酸イフェンプロジル 529


酒石酸エルゴタミン 452酒石酸キタサマイシン 1485酒石酸プロチレリン 721酒石酸レバロルファン 570酒石酸レバロルファン注射液 570ショウキョウ 927ショウキョウ末 927硝酸イソソルビド 558硝酸イソソルビド錠 558硝酸銀 746硝酸銀点眼液 1041硝酸チアミン 795, 1532硝酸ナファゾリン 636硝酸ミコナゾール 625常水 1076ショウズク 885消毒用エタノール 916消毒用フェノール 1010消毒用フェノール水 1012ショウマ 896ジョサマイシン 559, 1478シンイ 1552親水軟膏 940親水ワセリン 1005診断用クエン酸ナトリウム液 754シンフィブラート 746, 1524

ス

水酸化カリウム 1018水酸化カルシウム 877水酸化ナトリウム 1056スクラルファート 766ステアリルアルコール 1059ステアリン酸 1059ステアリン酸エリスロマイシン 454ステアリン酸カルシウム 881ステアリン酸ポリオキシル 40 1016ステアリン酸マグネシウム 969スピロノラクトン 765スルチアム 779スルバクタムナトリウム 768スルピリド 776スルピリン 776スルピリン注射液 777スルファジアジン銀 769スルファメチゾール 770スルファメトキサゾール 770スルファモノメトキシン 771スルフイソキサゾール 773スルフィンピラゾン 772スルフィンピラゾン錠 773スルベニシリンナトリウム 769,

1528スルホブロモフタレインナトリウム

774スルホブロモフタレインナトリウム注

射液 775

セ

成人用沈降ジフテリアトキソイド

912

精製水 1079精製ゼラチン 924精製セラック 1049精製脱脂綿 851精製デヒドロコール酸 395精製白糖 1060精製ラノリン 963生理食塩液 752石油ベンジン 1006セクレチン 745セスキオレイン酸ソルビタン 1058セタノール 894セッコウ 937セネガ 1043セネガシロップ 1044セネガ末 1044セファクロル 314, 1403セファゾリンナトリウム 319セファゾリンナトリウム水和物 319セファトリジンプロピレングリコール

318セファドロキシル 314セファピリンナトリウム 317セファマンドールナトリウム 317,

1407セファレキシン 315セファロチンナトリウム 317, 1406セファロリジン 317, 1405セフィキシム 329セフォキシチンナトリウム 336,

1419セフォジジムナトリウム 1412セフォタキシムナトリウム 335,

1413セフォテタン 335, 1415セフォペラゾンナトリウム 332セフジトレンピボキシル 325セフジニル 324セフスロジンナトリウム 340セフタジジム 342セフチゾキシムナトリウム 345セフチブテン 344, 1426セフテラムピボキシル 344, 1425セフトリアキソンナトリウム 346,

1427セフピラミドナトリウム 338, 1421セフブペラゾンナトリウム 322,

1409セフポドキシムプロキセチル 1422セフミノクスナトリウム 332セフメタゾールナトリウム 331セフラジン 339セフロキサジン 340, 1424セフロキシムアキセチル 347, 1429セフロキシムナトリウム 348ゼラチン 923センキュウ 899センキュウ末 899センコツ 983センソ 1071, 1563センナ 1044, 1560センナ末 1046, 1561センブリ 1063, 1562

センブリ・重曹散 1065, 1563センブリ末 1064, 1562

ソ

ソウジュツ 869ソウジュツ末 869ソウハクヒ 981, 1554ソヨウ 1004

タ

ダイオウ 1026, 1558ダイオウ末 1026, 1558ダイズ油 1059タイソウ 958胎盤性性腺刺激ホルモン 934タクシャ 860タクシャ末 860脱脂綿 851タルク 1065炭酸カリウム 1018炭酸水素ナトリウム 749炭酸水素ナトリウム注射液 750,

1524炭酸ナトリウム 1055炭酸マグネシウム 582炭酸リチウム 578単シロップ 1052ダントロレンナトリウム 392単軟膏 1052タンニン酸 782タンニン酸アルブミン 227タンニン酸ジフェンヒドラミン 427タンニン酸ベルベリン 272

チ

チアマゾール 791チアマゾール錠 792チアミラールナトリウム 796チアントール 1067チオテパ 799チオペンタールナトリウム 797,

1532チオ硫酸ナトリウム 762チオ硫酸ナトリウム注射液 762チカルシリンナトリウム 801チクセツニンジン 998チクセツニンジン末 998, 1555窒素 982チニダゾール 804チモ 864チモール 1069注射用アモバルビタールナトリウム

242注射用塩化アセチルコリン 222注射用塩化スキサメトニウム 781注射用塩酸ヒドララジン 513注射用血清性性腺刺激ホルモン 937注射用コハク酸プレドニゾロンナトリ

ウム 705, 1513注射用水 1078


注射用胎盤性性腺刺激ホルモン 936注射用チアミラールナトリウム 796注射用チオペンタールナトリウム

798注射用ファモチジン 470, 1464注射用フェニトインナトリウム 686注射用硫酸ビンブラスチン 841チョウジ 898チョウジ末 898チョウジ油 898チョウトウコウ 1563チョレイ 1061チョレイ末 1061チンク油 1085沈降ジフテリア破傷風混合トキソイド

913沈降精製百日せきジフテリア破傷風混

合ワクチン 1005沈降精製百日せきワクチン 1005沈降炭酸カルシウム 297沈降破傷風トキソイド 1066沈降はぶトキソイド 938沈降 B型肝炎ワクチン 938チンピ 898

ツ

ツバキ油 882

テ

テイコプラニン 783, 1530D-ソルビトール 763D-ソルビトール液 764D-マンニトール 587D-マンニトール注射液 588低置換度ヒドロキシプロピルセルロー

ス 942テオフィリン 791, 1531テガフール 782, 1530デキサメタゾン 397デキストラン硫酸ナトリウムイオウ

5 401デキストラン硫酸ナトリウムイオウ

18 402デキストラン 40 398デキストラン 40注射液 399デキストラン 70 400デキストリン 906デスラノシド 396デスラノシド注射液 397デヒドロコール酸 394デヒドロコール酸注射液 396テレビン油 1073天然ケイ酸アルミニウム 233テンマ 1550テンモンドウ 1546

ト

トウガラシ 882, 1546トウガラシ末 883, 1546トウガラシ・サリチル酸精 884

トウガラシチンキ 883トウキ 956トウキ末 956トウニン 1002トウニン末 1002トウヒ 875トウヒシロップ 994トウヒチンキ 995トウモロコシデンプン 903トウモロコシ油 903トコフェロール 807, 1534トコン 953トコンシロップ 955トコン末 954トシル酸スルタミシリン 778トチュウ 1548トフィソパム 812トブラマイシン 807, 1533トラガント 1072トラガント末 1072トラザミド 813トラネキサム酸 817トラピジル 818トリアムシノロン 819, 1535トリアムシノロンアセトニド 820トリアムテレン 821トリクロホスナトリウム 823トリクロホスナトリウムシロップ

824トリクロルメチアジド 822トリコマイシン 823, 1536トリメタジオン 827トリメタジオン錠 827トルナフタート 815トルナフタート液 816トルブタミド 814トルブタミド錠 814トレピブトン 819トロピカミド 828ドロペリドール 435, 1456トロンビン 1068豚脂 964

ナ

ナイスタチン 656ナタネ油 1024ナドロール 633ナファゾリン・クロルフェニラミン液

981ナプロキセン 636ナリジクス酸 634

ニ

ニガキ 1013ニガキ末 1013ニコチン酸 644ニコチン酸アミド 643ニコチン酸注射液 645ニコチン酸トコフェロール 810ニコモール 642ニコモール錠 643

二酸化炭素 312ニセリトロール 641ニトラゼパム 646ニトログリセリン錠 647ニフェジピン 645日本脳炎ワクチン 956乳酸 959乳酸カルシウム 300乳糖 960尿素 835ニンジン 927ニンジン末 929

ノ

濃塩化ベンザルコニウム液 50 267濃グリセリン 504, 1469ノスカピン 654ノルエチステロン 650ノルエピネフリン 649, 1498ノルエピネフリン注射液 650, 1499ノルゲストレル 652ノルゲストレル・エチニルエストラジ

オール錠 652, 1499ノルフロキサシン 651

ハ

バイモ 1549白色セラック 1049白色軟膏 1080白色ワセリン 1005白糖 1061バクモンドウ 987バクロフェン 260バクロフェン錠 261バシトラシン 1390バソプレシン注射液 837ハチミツ 938ハッカ 974ハッカ水 975ハッカ油 975パップ用複方オウバク散 1009, 1556パニペネム 667ハマボウフウ 930パモ酸ヒドロキシジン 524パモ酸ピランテル 722パラアミノサリチル酸カルシウム

301パラアミノサリチル酸カルシウム顆粒

302パラオキシ安息香酸エチル 916パラオキシ安息香酸ブチル 876パラオキシ安息香酸プロピル 1021パラオキシ安息香酸メチル 976パラフィン 999パラホルムアルデヒド 671バルビタール 263バルプロ酸ナトリウム 762パルミチン酸クロラムフェニコール

1432パルミチン酸レチノール 733, 1519バレイショデンプン 1019


ハロキサゾラム 509, 1472ハロタン 508ハロペリドール 507パンクレアチン 999ハンゲ 1013絆創膏 858パンテチン 669パントテン酸カルシウム 301

ヒ

ピコスルファートナトリウム 758ビサコジル 280ビサコジル坐剤 280, 1396ビタミン A油 1074, 1564ビタミン A油カプセル 1075, 1565ヒトインスリン（遺伝子組換え） 537人全血液 1080人免疫グロブリン 949ヒドロキシプロピルセルロース 940ヒドロキシプロピルメチルセルロース

フタレート 947ヒドロキシプロピルメチルセルロース

2208 943ヒドロキシプロピルメチルセルロース

2906 944ヒドロキシプロピルメチルセルロース

2910 945ヒドロクロロチアジド 515, 1473ヒドロコルチゾン 516ヒドロコルチゾン・ジフェンヒドラミ

ン軟膏 939ピペミド酸三水和物 689, 1508ピペラシリンナトリウム 690, 1508ヒベンズ酸チペピジン 804, 1533ヒベンズ酸チペピジン錠 806ビホナゾール 278ヒマシ油 888ピマリシン 1507ヒメクロモン 525ビャクシ 864ビャクジュツ 868ビャクジュツ末 868氷酢酸 856ピラジナミド 723, 1515ピラルビシン 1509ピレノキシン 693ピロ亜硫酸ナトリウム 1057ピロキシリン 1023ピロールニトリン 1516ビワヨウ 1552ピンドロール 688ビンロウジ 866, 1545

フ

ファノバリン・マグネシア散 1557ファモチジン 469ファモチジン散 469ファモチジン錠 471ファロペネムナトリウム 472フィトナジオン 687, 1507フェナセチン 680

フェニトイン 685フェニトイン散 686フェニトイン錠 686フェニルブタゾン 684フェネチシリンカリウム 1505フェノバリン・マグネシア散 1012フェノバルビタール 681フェノバルビタール散 10 681フェノール 1010フェノール・亜鉛華リニメント 1011フェノール水 1012フェノールスルホンフタレイン 682フェノールスルホンフタレイン注射液

682フェンブフェン 473複方アクリノール・チンク油 857複方オキシコドン・アトロピン注射液

996複方オキシコドン注射液 995複方サリチル酸精 1033複方サリチル酸メチル精 977複方ジアスターゼ・重曹散 907複方ダイオウ・センナ散 1028複方チアントール・サリチル酸液

1067複方ビタミン B散 1075複方ヨード・グリセリン 950複方ロートエキス・ジアスターゼ散

1037複方ロートエキス・タンニン坐剤

1041, 1560複方ロートエキス・タンニン軟膏

1039, 1560ブクリョウ 1017ブクリョウ末 1017フシジン酸ナトリウム 1525ブスルファン 293ブドウ酒 1081ブドウ糖 503ブドウ糖注射液 503ブフェキサマク 289ブフェキサマク軟膏 290ブフェキサマク乳剤性軟膏 289フマル酸クレマスチン 368フマル酸ケトチフェン 1483フマル酸ホルモテロール 492ブメタニド 291プラステロン硫酸ナトリウム 760プラゼパム 700プラゼパム錠 701プラノプロフェン 699フラビンアデニンジヌクレオチドナト

リウム 475プリミドン 706, 1514フルオキシメステロン 486フルオシノニド 482フルオシノロンアセトニド 481フルオレセインナトリウム 483フルオロウラシル 485フルオロメトロン 484フルジアゼパム 479フルシトシン 478フルニトラゼパム 480

フルラゼパム 487フルラゼパムカプセル 488フルルビプロフェン 489, 1466プレドニゾロン 701プレドニゾロン錠 702フロクタフェニン 477, 1465プログルミド 715プロゲステロン 714プロゲステロン注射液 714フロセミド 498プロタミンインスリン亜鉛水性懸濁注

射液 544プロチオナミド 720プロチレリン 720プロテイン銀 1051プロテイン銀液 1051プロピオン酸ジョサマイシン 559,

1480プロピオン酸テストステロン 788プロピオン酸テストステロン注射液

788プロピオン酸ドロスタノロン 435,

1456プロピオン酸ドロスタノロン注射液

436, 1456プロピオン酸ベクロメタゾン 264プロピルチオウラシル 718プロピルチオウラシル錠 718プロピレングリコール 1022フロプロピオン 478プロベネシド 706プロベネシド錠 707ブロマゼパム 284ブロムワレリル尿素 287フロモキセフナトリウム 478, 1465粉末セルロース 892

ヘ

ベタメタゾン 274ヘパリンナトリウム 510ヘパリンナトリウム注射液 511ベラドンナエキス 871ベラドンナコン 872ペルフェナジン 675ペルフェナジン錠 676ベンジルアルコール 874ベンジルペニシリンカリウム 270,

1393ベンジルペニシリンベンザチン

1392ベンズブロマロン 268, 1391ペンタゾシン 672ベントナイト 873ペントバルビタールカルシウム 673

ホ

ボウイ 1052, 1562ボウコン 949, 1551ホウ砂 750ホウ酸 284抱水クロラール 350


ボウフウ 1034ホスフェストロール 493ホスフェストロール錠 494ホスホマイシンカルシウム 494ホスホマイシンナトリウム 496ボタンピ 979ボタンピ末 980ポビドン 1020ポビドンヨード 699, 1513ホミカ 983ホミカエキス 984ホミカエキス散 984ホミカチンキ 985ポリスチレンスルホン酸カルシウム

303, 1399ポリスチレンスルホン酸ナトリウム

759ポリソルベート 80 1017ホリナートカルシウム 299ホルマリン 919ホルマリン水 920ボレイ 997ボレイ末 998

マ

マイトマイシン C 629, 1496マオウ 914マーキュロクロム 604マーキュロクロム液 605マクリ 908マクロゴール軟膏 968マクロゴール 400 965マクロゴール 1500 966マクロゴール 4000 967マクロゴール 6000 967マクロゴール 20000 968マシニン 1551麻酔用エーテル 462マルトース 586マレイン酸エルゴメトリン 450マレイン酸エルゴメトリン錠 451マレイン酸エルゴメトリン注射液

451マレイン酸クロルフェニラミン 357d-マレイン酸クロルフェニラミン

359マレイン酸クロルフェニラミン散

358マレイン酸クロルフェニラミン錠

359マレイン酸クロルフェニラミン注射液

357マレイン酸トリメブチン 1537マレイン酸プロクロルペラジン 713マレイン酸プロクロルペラジン錠

713マレイン酸ペルフェナジン 677マレイン酸ペルフェナジン錠 678マレイン酸メチルエルゴメトリン

615マレイン酸メチルエルゴメトリン錠

615

マレイン酸レボメプロマジン 572D-マンニトール注射液 1492

ミ

ミグレニン 627, 1495ミコナゾール 624ミツロウ 871ミデカマイシン 626ミョウバン水 861

ム

無晶性インスリン亜鉛水性懸濁注射液

542無水アンピシリン 246, 1384無水エタノール 915無水カフェイン 295無水クエン酸 367, 1438無水乳糖 961無水リン酸水素カルシウム 879ムピロシンカルシウム水和物 631

メ

メキタジン 602メグルミン 974メコバラミン 590メシル酸ガベキサート 500メシル酸カモスタット 304メシル酸ジヒドロエルゴタミン 418メシル酸ジヒドロエルゴトキシン

413, 1450メシル酸デフェロキサミン 393メシル酸ブロモクリプチン 286メシル酸ベタヒスチン 272メストラノール 606メダゼパム 591メチクラン 619, 1493メチラポン 623メチルジゴキシン 620メチルセルロース 978メチルテストステロン 618メチルテストステロン錠 619メチルドパ 612メチルドパ錠 613メチルプレドニゾロン 616メチル硫酸ネオスチグミン 637メチル硫酸ネオスチグミン注射液

638滅菌ガーゼ 854滅菌精製水 1079滅菌精製脱脂綿 852滅菌脱脂綿 852メトキサレン 611メトクロプラミド 621メトトレキサート 610, 1493メトロニダゾール 622メナテトレノン 598, 1492メピチオスタン 600, 1492メフェナム酸 592メフルシド 592メフルシド錠 593

メルカプトプリン 603メルファラン 597メロペネム三水和物 605dl-メントール 975l-メントール 976

モ

モクツウ 859モッコウ 1034モノステアリン酸アルミニウム 861モノステアリン酸グリセリン 931モルヒネ・アトロピン注射液 978,

1553

ヤ

焼セッコウ 938ヤクチ 875薬用石ケン 973薬用炭 972ヤシ油 899

ユ

ユウタン 869ユーカリ油 917輸血用クエン酸ナトリウム注射液

753ユビデカレノン 832

ヨ

ヨウ化エコチオパート 438ヨウ化オキサピウム 657ヨウ化カリウム 697ヨウ化人血清アルブミン (131I)注射液

546ヨウ化ナトリウム 755ヨウ化ナトリウム (123I)カプセル

755ヨウ化ナトリウム (131I)液 756ヨウ化ナトリウム (131I)カプセル

755ヨウ化ヒプル酸ナトリウム (131I)注射

液 756葉酸 490, 1466葉酸錠 491葉酸注射液 491ヨウ素 546ヨクイニン 900ヨクイニン末 900ヨーダミド 545ヨーダミドナトリウムメグルミン注射

液 597ヨード・サリチル酸・フェノール精

952ヨードチンキ 949ヨードホルム 546

ラ

ラウリル硫酸ナトリウム 1056


ラウロマクロゴール 964酪酸ヒドロコルチゾン 518酪酸リボフラビン 734ラクツロース 566ラクトビオン酸エリスロマイシン

1462ラタモキセフナトリウム 569, 1486ラッカセイ油 1002ラナトシド C 567ラナトシド C錠 567

リ

リオチロニンナトリウム 576リオチロニンナトリウム錠 577,

1490リドカイン 574リドカイン注射液 575リファンピシン 737, 1521リボフラビン 733リボフラビン散 734リュウコツ 965硫酸亜鉛 846硫酸亜鉛点眼液 1086硫酸アストロマイシン 255, 1387硫酸アトロピン 255硫酸アトロピン注射液 256硫酸アミカシン 238硫酸アルベカシン 247, 1386硫酸アルミニウムカリウム 862硫酸イセパマイシン 552硫酸エンビオマイシン 1457硫酸オルシプレナリン 656硫酸カナマイシン 563, 1482一硫酸カナマイシン 1481硫酸カリウム 1019硫酸キニジン 725硫酸キニーネ 728硫酸グアネチジン 507硫酸ゲンタマイシン 502, 1468硫酸コリスチン 1444硫酸サルブタモール 740硫酸シソマイシン 747, 1524硫酸ジベカシン 404, 1447

硫酸ストレプトマイシン 766, 1527硫酸セフォセリス 334硫酸セフピロム 338硫酸鉄 474硫酸テルブタリン 786硫酸ネチルマイシン 630硫酸バメタン 262硫酸バリウム 264硫酸ビンクリスチン 842硫酸ビンブラスチン 840硫酸フラジオマイシン 497, 1467硫酸ブレオマイシン 283, 1398硫酸プロタミン 719硫酸プロタミン注射液 719硫酸ベカナマイシン 265, 1390硫酸ペプロマイシン 675, 1503硫酸ペンブトロール 672硫酸ポリミキシン B 694, 1511硫酸マグネシウム 585硫酸マグネシウム水 970硫酸マグネシウム注射液 586, 1492硫酸ミクロノマイシン 626, 1494硫酸リボスタマイシン 737, 1520リュウタン 957リュウタン末 957, 1551流動パラフィン 1000リョウキョウ 1544リンゲル液 1028リン酸クリンダマイシン 369, 1440リン酸コデイン 382リン酸コデイン散 1 382, 1442リン酸コデイン散 10 383, 1442リン酸コデイン錠 384, 1443リン酸ジヒドロコデイン 417リン酸ジヒドロコデイン散 1 417,

1449リン酸ジヒドロコデイン散 10

418, 1450リン酸ジメモルファン 421リン酸水素カルシウム 878リン酸水素ナトリウム 907リン酸二水素カルシウム 880リン酸ヒドロコルチゾンナトリウム

519, 1474

リン酸ピペラジン 692リン酸ピペラジン錠 692リン酸ベタメタゾンナトリウム 276,

1395リン酸リボフラビンナトリウム 735リン酸リボフラビンナトリウム注射液

736

レ

レセルピン 729, 1518レセルピン散 0.1 731レセルピン錠 731レセルピン注射液 730レボチロキシンナトリウム 572レボチロキシンナトリウム錠 573レボドパ 571レンギョウ 920

ロ

ロキシスロマイシン 738, 1522ロキソプロフェンナトリウム 580ロキタマイシン 737ロジン 1029ロートエキス 1035ロートエキス・アネスタミン散 1037,

1559ロートエキス・カーボン散 1037ロートエキス・タンニン坐剤 1039ロートエキス・パパベリン・アネスタミ

ン散 1038ロートエキス散 1036ロートコン 1040ロラゼパム 579

ワ

ワイル病秋やみ混合ワクチン 1080ワルファリンカリウム 843ワルファリンカリウム錠 843

supplement i to the japanese pharmacopoeia fourteenth edition

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