september 2010 - uni-giessen.de · september 2010 the versatility of hptlc – from bio analysis to...

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The versatility of HPTLC – From Bio Analysis to the Detection of Pollutants in Water

AMD chromatogram of Stratum corneum lipids

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CAMAG BIBLIOGRAPHY SERVICE

No. 105, September 2010CAMAG Bibliography Service Planar Chromatography Edited by Gerda Morlock [email protected] published by CAMAG Switzerland

CAMAG (Switzerland) Sonnenmattstr. 11 • CH-4132 Muttenz 1 Tel. +41 61 4673434 • Fax +41 61 4610702 [email protected] • www.camag.com

CAMAG Scientific Inc. (USA) 515 Cornelius Harnett Drive Wilmington, NC 28401 Phone 800 334 3909 • Fax 910 343 1834 [email protected] • www.camagusa.com

IN THIS ISSUE

Procedures, applicationsTLC/HPTLC-ELSD-MS coupling ...... 2–4

Determination of the glycoalkaloids -solanine and -chaconine in potatoes at different steps of potato processing ..................... 5–6

1H-Benzotriazole and tolyltriazole in the aquatic environment..................... 7–9

Optimization of an AMD 2 method for determination of stratum corneum lipids ............. 10–12

Determination of additives in plastic foils ........................... 13–15

Products featured in this issue

TLC Scanner 4 .................................. 4

Automated Multiple Development (AMD 2) ....... 11

TLC-MS Interface ........................... 16

Column: Know CAMAGSeminar, held by CAMAG Berlin at the University of Zittau/Görlitz ...... 8

Printed on FSC/PEFC-certified paper

Planar Chromatography in Practice

TLC/HPTLC-ELSD-MS coupling

Mr. François Bretin and Dr. Francis Maquin (right)

The Lead Generation to Candidate Realization (LGCR) platform identifies small molecule leads and progresses them up to registration. LGCR Ana-lytical Sciences (AnSci) is a new department in Sanofi-Aventis R&D, which is dedicated to support Business Divisions and Therapeutics Units. For LGCR-AnSci, Dr. Maquin* and F. Bretin in the research center in Vitry-sur-Seine use various analytical techniques (further locations are in Strasbourg, Chilly-Mazarin, Toulouse and Montpellier).

IntroductionMS coupled to chromatography supports the chemist’s decision for compound identification or follow-up in a synthesis mixture. Therefore, HPLC/UPLC-MS is mainly used, but TLC/HPTLC is also widely applied by the chemists as a rapid and reliable method to follow reaction processes.

This is especially true for cases, when compounds remain on the column, due to high polarity or weak solubility, or when analyte detection is poor (no chromophore). In such cases, the HPTLC/MS platform with the new TLC-MS Interface is ideal for structural analysis. The TLC-MS Interface was complemented by a MS soft-ware controlled valve, adding some automation to the process, and by an Evaporative Light Scattering Detector (ELSD). The latter enables the analyst to distinguish between compounds that were not eluted from the plate from those that were not sufficiently ionized for MS detection.

Sample preparationThe samples were taken directly from the reactor and diluted with an ap-propriate solvent, usually an organic solvent of medium polarity.

Standard preparationEducts or known intermediate products were dissolved and diluted with an appropriate solvent.

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Chromatogram layersTLC and HPTLC plates silica gel 60 F254 10 x 20 and 10 x 10 cm, respectively. If required, the plate size was reduced with the Smartcut.

Sample applicationManually with disposable micropipettes of 5 to 20 µL volumes.Note: The Nanomat 4 is recommended to ensure precise positioning without damage of the layer. Especially for polar extracts, small volumes should be applied (<2 µL to obtain sharp start zones).

ChromatographyIn a twin-through chamber, e.g., with mixtures of methanol and dichloromethane/ethyl acetate, or ethyl acetate and heptane/cyclohexane; the ratios depend on the compound mixtures.

DerivatizationCompounds with neither UV/Vis-activity nor native fluorescence can be derivatized by non-destruc-tive derivatization reagents, e.g., the primuline or berberine reagents for lipophilic compounds, and directly eluted with the TLC-MS interface into the MS. However, for destructive derivatizations, e.g., based on strong acidic carbonization reactions, on both plate sides, the outer track was cut, deriva-tized, and the respective bands were marked by extrapolation.

Recording of MS spectraThe flow rate of the eluent (methanol – water 95:5, 0.4 mL/min) was split by a tee, and 0.2 mL/min were pumped to the MS (Micromass ZQ, Waters) and 0.2 mL/min to the ELSD (Sedere Sedex 85 LT). The TLC-MS Interface was equipped with either a round or oval elution head and connected via an external automated valve (MXP7900-000, Rheodyne) to the pump (Alliance 2695, Waters) and ESI-MS. In the transfer tube to the detectors, an inline filter was integrated (frit porosity 0.2 µm, A 356/504, Upchurch IDEX). The recording of mass spectra was performed in the positive/negative electrospray mode; for evaluation the MS software (Mass Lynx V4.0/Open Lynx, Waters) was used.

TLC/HPTLC-ELSD-MS system configuration

Results and discussionIn our experiments, a lot of organic structures were not detected by DAD, so the capability of detection of the TLC/HPTLC-DAD-MS configuration was im-proved by substituting the DAD with the ELSD. As universal detector, the ELSD confirms more reliably the elution of compounds that are not UV/Vis-active.

Once the elution head was lowered onto the zone of interest and an ID number was entered in the software, valve switching was effected remotely, followed by an automatic acquisition of both the MS and ELSD signal. This minor automation resulted in the zones being eluted into the detector, and then the eluent being subsequently switched back to bypass (waste). The following protocol made cross contamination, which of course is dependent on the substance structure and zone concentration, less likely. For the given connecting tube length and internal diameter, the external valve was set 10 s in bypass, then 1.5 min in elution mode and finally 15 s in bypass. For a high throughput, the elution time was set below one minute.

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Elution profiles obtained by ELSD and ESI-MS (top) and mass spectrum of the eluted zone (bottom)

An intensive use of the TLC/HPTLC-ELSD-MS instru-mentation requires frit cleaning twice a week. The requirement of a cleaning cycle is indicated by an increased pump pressure likely from clogging of the inline filter frit (> 10 MPa).

To conclude, our experience underlines the interest in HPTLC/TLC-MS in an advanced analytical environ-ment. The automatization, reliability of the data, and speed were the convincing arguments, which fully comply with our routine analytical needs.

Further information is available on request from the authors.

Contacts• Dr. Francis Maquin and Mr. François Bretin, Sanofi-Aventis, Centre de Recherche, 13, quai Jules Guesde, 94403 Vitry- sur-Seine cedex, France, [email protected]

• Mr. Pierre Bernard-Savary, Chromacim SAS, Pommiers la Placette, France

• Mr. Henri Gangloff, Sedere SA, Alfortville, France

• Mrs. Véronique de Nailly, BCP Instruments, Irigny, France

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CAMAG TLC Scanner 4

Classical densitometry uses monochromatic light in the form of a slit of selectable length and width to scan the tracks of a chromatogram, measuring the diffusely reflected light. The TLC Scanner 4 – the successor of TLC Scanner 3 – is the most advanced workstation for densitometric evaluation of Thin-Layer Chromatograms currently available.

Analysts can now benefit from a wider spec-tral range for measurements from 190 to 900 nm. The optimized positioning stage and its open access allow for robust use.

For the screening of water samples in this application, densitometric determination is performed by the multi-wavelength scan covering the wavelength range between 190 and 300 nm. In combination with AMD the whole polarity range of UV-active compounds is detected. This procedure is proven to be helpful in routine search for potential water contaminants.

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Determination of the glycoalkaloids -solanine and -chaconine in potatoes at different steps of potato processing

Dr. Jens Mäder, Prof. Dr. Lothar W. Kroh

Prof. Dr. Kroh*, TU Berlin, and his group, concen- trate on carbohydrate chemistry and food analysis, especially in Maillard reaction chemistry, carameliza-tion and melanoidin formation. A further research field of both authors is the characterization of bioactive compounds of processed and unproc-essed food. Dr. Mäder is now head of product development at Milchwerke Mittelelbe, Stendal. This study [1] was performed by Dr. Mäder, Ms Hanschen and Ms Zietz (both not pictured) dur-ing their graduation.

IntroductionThe potato as a plant of the Solanaceae family contains bioactive steroidal alkaloids, which are de-terrent to herbivores and pathogens and reported to be membrantoxic, embryotoxic and teratogenic for mammalians. They influence the flavor of fresh and processed potatoes from a bitter taste toward a burning sensation at higher concentrations [2]. At the moment no limit values are fixed in Europe, and potatoes, as the most important food source of glycoalkaloids, are regarded as safe for humans if they do not contain more than 200 mg total al-kaloids per kg of fresh weight. Both main alkaloids -solanine and -chaconine exist at high levels in the tuber peel in a ratio of 1:2 to 1:7.

Chemical structure of the potato alkaloids -solanine and -chaconine

The potato is one of the world’s most important food crops. Quality-management systems demand food safety and traceability as they often follow the from farm to fork policy of regulatory authorities. Therefore, easy to use, inexpensive, and reliable analytical methods are needed to evaluate the content of toxicologically relevant compounds like glycoalkaloids after harvest and during potato stor-age and processing.

HPTLC is especially suited for effective and rapid determination of alkaloids [4] because up to 16 samples can be simultaneously separated without any matrix influence on the quality of the results. Consequently, all processing stages and by-products from one batch can be moni-tored and traced simultaneously within one analysis on one plate.

Sample preparationSamples were collected at different stages of the production chain in a commercial potato flake process. After fourfold extraction of the lyophilized, ground potato powder with methanol - acetic acid 99:5, the extracts were concentrated and directly applied.

Standard solutions5 mg each of -solanine and -chaconine were dis-solved in 25 mL methanol – acetic acid 99:1, mixed 1:1 and diluted 1:20.

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Chromatogram layerHPTLC plate silica gel 60 (Merck), 20 x 10 cm

Sample applicationBandwise with Automatic TLC Sampler 4, band length 5 mm, track distance 9.2 mm, distance from the lower edge 8 mm, application speed 70 nL/s, application volume 2–22 µL

ChromatographyIn the Horizontal Developing Chamber after 15 min chamber saturation with dichloromethane – metha-nol – ammonia (2.5 %) 70:30:4.4, migration dis-tance of 75 mm from the lower edge of plate

Post-chromatographic derivatizationWith the Chromatogram Immersion Device III the plate was dipped after 25 min drying time at 90 °C into the Carr-Price reagent (70 g SbCl3 is dissolved in 250 mL of a mixture of acetic acid – dichloro-methane 1:3), followed by heating on the TLC Plate Heater at 110 °C for 3 min.

DensitometryAbsorption measurement at 560 nm with TLC Scanner 3 and winCATS software within 15 min after derivatization, before the glycoalkaloid zones change the color from red to violet.

Results and discussionThe densitogram clearly shows the good separation of -solanine and -chaconine in the different sam-ple extracts. The extremely sensitive and selective coloring of both glycoalkaloids by dipping into the Carr-Price reagent allowed a limit of detection and quantification of 5–15 (depending on the matrix) and 30 ng/band, respectively, with a recovery rate between 94 and 105 %. Linear calibration was very satisfactory in the range of 30 to 700 ng/band (r = 0.9998, sdv = 2.5 %), polynomial even up to 1500 ng/ band (r = 0.9999, sdv = 1.5 %).

In conclusion, this method ideally complies with the analytical demands mentioned in the introduction.

Densitogram of standard substances (track 1–6) and sample extracts of different steps of potato processing (track 7–21)

Further information is available from the author on request.

*Prof. Dr. Lothar W. Kroh, lnstitut für Lebensmitteltechnologie und Lebensmittelchemie, TU Berlin, Gustav-Meyer-Allee 25, 13355 Berlin, [email protected]

Literature

[1] J. Mäder et al. J Planar Chromatogr 22 (2009) 43.

[2] S.L. Sinden , K.L. Deahl and B.B. Aulenbach, J Food Sci 41 (1976) 520.

[3] A.J.A. Essers et al. Environ Toxicol Phar 5 (1998) 155.

[4] J. Mäder, H. Rawel and L.W. Kroh, J Agric Food Chem 57 (2009) 6292.

-solan

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aconine

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continuation on page 9


1H-Benzotriazole and tolyltriazole in the aquatic environment

Dr. Walter H. Weber

Dr. Wolfram Seitz, Stefan C. Weiss, Alexander Müller and Dr. Wolfgang Schulz (from left to right)

Lead by Dr. Weber* the laboratory for operation con-trol and research of Zweckverband Landeswasserver-sorgung (LW) in Langenau, Germany, is active in the analysis of drinking water as well as in the monitoring of ground and waste water from the water protection area Donauried-Hürbe and of surface water from the surrounding area. HPTLC, preferably with automated multiple development (AMD), and online hyphenation of HPTLC-MS are part of the methods applied in routine analysis [1–3].

IntroductionWithin the scope of a monitoring program of raw water resources used by LW, comprehensive chemical, physico-chemical and microbiological analyses are performed regularly. In addition non-target screening analyses for the detection of not yet considered contaminants or unknown substances were performed.

In one of the HPTLC/AMD screening tests of ex-tracts from secondary effluents and of surface and ground water samples the resulting chroma-tograms showed a zone which could not be identi-fied as any of the substances known from target analysis at LW.

Sample preparationAnalyte enrichment from a 100 mL water sample was done by solid phase extraction (SPE) using 0.2 g of Iso-lute ENV+ sorbent at pH 3 or 7. The SPE sorbent was conditioned consecutively with 6 mL each of n-hexane, acetone, methanol and water (pH = 3 or 7). After drying 6 mL of methanol were used for elution and after eva-

poration to dryness the residue was taken up in 200 µL of methanol.

LayerHPTLC plates Lichrospher F254, 20 x 10 cm (Merck) were pre-cleaned with 2-propanol and dried for 30 min at 120 °C on the TLC Plate Heater.

Sample applicationWith Automatic TLC Sampler (ATS4) as area of 6 x 3 mm

ChromatographyIn the AMD 2 system with a 25-step gradi-ent starting in isocratic mode with 5 steps acetonitrile – dichloromethane 1:1 for focusing, followed by 15 steps from acetonitrile – dichlo-romethane 1:1 to dichloromethane and then in 5 steps from dichloromethane – n-hexane 4:1 to n-hexane. The final migration distance was 80 mm.

Densitometric evaluationMulti-wavelength scan at 190, 200, 220, 240, 260, 280 and 300 nm with the TLC Scanner and winCATS software

DocumentationUnder UV 254 nm with the TLC Visualizer

Results and discussionDuring routine screening of extracts from secondary effluents as well as of surface and ground water samples a high peak was ob-served in the chromatograms. To further inves-tigate this peak the unknown zone was eluted from the HPTLC plate by means of the TLC-MS Interface and analyzed by Nano LC followed by QTOF-MS (ESI+). The analysis revealed that the zone consisted of two substances, peak A at RT = 3.2 min and peak B at RT = 3.4 min.

Dear friends

This CBS issue has been delayed a bit, but in my de-fense, I would like to men-tion that during the hot fi-nal phase of the September issue, I was co-organizing the 39th German Food Congress. It was held in Stuttgart-Hohenheim this year and attended by over 600 participants. I think the congress was quite fruitful for the progress of HPTLC in food analysis, especially due to a workshop for the Young Food Chemists.

It was noted that many international compa-nies reported an increase in the need for more food analyses, for which HPTLC offers excel-lent preconditions. There is great pent-up de-mand in countries like China, India, Thailand, and other countries where food safety is more and more forced by law.

Due to the increased demand for food analysis, three articles in this CBS relate to this field. By the way the study of food chemistry is a Ger-man unicum – there is no comparable study in other countries.

Please keep in mind that preparations for the HPTLC Symposium in Basel, 6–8. July 2011 proceed. Information (Call for Papers) can be found on the last yellow page. Do we meet in Basel? I would enjoy it!

Sincerely,

Gerda Morlock [email protected]

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CAMAG LITERATURDIENSTCAMAG BIBLIOGRAPHY SERVICEPLANAR CHROMATOGRAPHY

Liebe Freunde

Dieser CBS erscheint leider mit Verspätung. In der Zeit, in der die Bearbeitung der September-Ausgabe in das Endstadium rückt, war ich als Mitorganisator stark eingebunden in die Orga-nisation des 39. Deutschen Lebensmittelche-mikertages in Stuttgart-Hohenheim. Dieser er-wies sich für die Verbreitung der HPTLC in der Lebensmittelanalytik als erfreulich ergiebig, vor allem im Workshop der jungen Lebensmittel-chemiker.

Erfreulich ist, dass viele international tätige Un-ternehmen einen stark ansteigenden Bedarf an Lebensmittelanalytik haben, für die die HPTLC vorzügliche Voraussetzungen bietet. Grossen Nachholbedarf auf diesem Gebiet haben Län-der wie China, Indien oder Thailand, in denen Lebensmittelsicherheit verstärkt gesetzlich ge-fordert wird.

Wegen der wachsenden Bedeutung der Le-bensmittelanalytik stammen drei Anwendungs-beiträge dieses CBS aus diesem Bereich im wei-teren Sinne. Übrigens ist der Studiengang Le-bensmittelchemie einzigartig in Deutschland - international gibt es nichts Vergleichbares.

Die Vorbereitungen zum HPTLC Symposium in Basel, 6.–8. Juli 2011, schreiten zügig voran. Informationen (Call for Papers) finden Sie auf der letzten gelben Seite. Sehen wir uns in Basel? Ich würde mich freuen!

Herzlichst

Gerda Morlock [email protected]

SEPTEMBER

2010

THE CBS CLASSIFICATION SYSTEM1. Reviews and books

a) Books on TLC b) Books containing one or several chapters on TLC c) Books containing frequent TLC information spread over several chapters of other information

2. Fundamentals, theory and general a) General b) Thermodynamics and theoretical relationship c) Relationship between structure and chrom. behaviour d) Measurement of physico-chemical and related values e) Optimization of solvent systems f) Validation of methods

3. General techniques (unless they are restricted to the application within one or two classification sections) a) New apparatus/techniques for sample preparation b) Separation material c) New apparatus for sample application/dosage d) New apparatus/techniques for chromatogram development e) New apparatus/techniques for pre- or post- chromatographic derivatization f) New apparatus/techniques for quantitative evaluation g) New apparatus/techniques for other TLC steps (distinguished from section 4)

4. Special techniques a) Automation of sample preparation/application b) Automation of complex chromatogram developing techniques c) Automation, computer application in quantitative chromatogram evaluation d) Combination of TLC with other chromatographic techniques e) Combination of TLC with other (non-chromatographic) techniques...MS, IR...etc.

5. Hydrocarbons and halogen derivatives a) Aliphatic hydrocarbons b) Cyclic hydrocarbons c) Halogen derivatives d) Complex hydrocarbon mixtures

6. Alcohols

7. Phenols

8. Substances containing heterocyclic oxygen a) Flavonoids b) Other compounds with heterocyclic oxygen

9. Oxo compounds, ethers and epoxides

10. Carbohydrates a) Mono- and oligosaccharides, structural studies b) Polysaccharides, mucopolysaccharides, lipopolysaccharides

11. Organic acids and lipids a) Organic acids and simple esters b) Prostaglandins c) Lipids and their constituents d) Lipoproteins and their constituents e) Glycosphingolipids (gangliosides, sulfatides, neutral glycosphingolipids)

12. Organic peroxides

13. Steroids a) Pregnane and androstane derivatives b) Estrogens c) Sterols d) Bile acids and alcohols e) Ecdysones and other insect steroid hormones

14. Steroid glycosides, saponins and other terpenoid glycosides

15. Terpenes and other volatile plant ingredients a) Terpenes b) Essential oils

16. Nitro and nitroso compounds

17. Amines, amides and related nitrogen compounds a) Amines and polyamines b) Catecholamines and their metabolites c) Amino derivatives and amides (excluding peptides)

18. Amino acids and peptides, chemical structure of proteins a) Amino acids and their derivatives b) Peptides and peptidic proteinous hormones

19. Proteins

20. Enzymes

21. Purines, pyrimidines, nucleic acids and their constituents a) Purines, pyrimidines, nucleosides, nucleotides b) Nucleic acids, RNA, DNA

22. Alkaloids

23. Other substances containing heterocyclic nitrogen a) Porphyrins and other pyrroles b) Bile pigments c) Indole derivatives d) Pyridine derivatives e) other N-heterocyclic compounds

24. Organic sulfur compounds

25. Organic phosphorus compounds (other than phospholipids)

26. Organometallic and related compounds a) Organometallic compounds b) Boranes, silanes and related non-metallic compounds c) Coordination compounds

27. Vitamins and various growth regulators (non-peptidic)

28. Antibiotics, Mycotoxins a) Antibiotics b) Aflatoxins and other mycotoxins

29. Pesticides and other agrochemicals a) Chlorinated insecticides b) Phosphorus insecticides c) Carbamates d) Herbicides e) Fungicides f) Other types of pesticides and various agrochemicals

30. Synthetic and natural dyes a) Synthetic dyes b) Chloroplasts and other natural pigments

31. Plastics and their intermediates

32. Pharmaceutical and biomedical applications a) Synthetic drugs b) Pharmacokinetic studies c) Drug monitoring d) Toxicological applications e) Plant extracts, herbal and traditional medicines f) Clinico-chemical applications and profiling body fluids

33. Inorganic substances a) Cations b) Anions

34. Radioactive and other isotopic compounds

35. Other technical products and complex mixtures a) Surfactants b) Antioxidants and preservatives c) Various specific technical products d) Complex mixtures and non-identified compounds

36. Thin-layer electrophoresis

37. Environmental analysis a) General papers b) Air pollution c) Water pollution d) Soil pollution

38. Chiral separations

XX. (abstract number underlined) refers to HPTLC related publication or application using HPTLC materials

CAMAGBIBLIOGRAPHYSERVICE No.105�

1. Reviews and books

105001 P.K.JAISWAL(CentralAgmarkLaboratory,Govt.ofIndia,NorthAmbazariRoad,Nagpur440010, India):High-performance thin-layerchromatography in foodanalysis.CBSPublishers&DistributorsPvtLtd,NewDehli,2010.Shortreviewonplanarchromatographyinfoodanalysis.Certainwell-knownapplicationsarementionedbutimportantpublicationsandrecentscientificdataislacking.

foodanalysis,HPTLC 1a

105002 MonikaJANICKA*,A.WASIK,J.NAMIESNIK(*GdanskUniversityofTechnology,ChemicalFaculty,DepartmentofAnalyticalChemistry,80-22�Gdansk,Poland,[email protected]):Analytical procedures for determination of cocaine and its metabolites in biological samples.TrAC29,209-224(2010).Thedangerouseffectsofcocaineonlivingorganismsandcurrentpro-blemsassociatedwithitswideuseandavailabilityhaveledtothedevelopmentofvariousana-lyticalprocedures.Thisreviewdiscussesanalyticalmethodsproposedduringthepasttenyearsfor theverificationof cocaineand itsmetabolites inbiological samples.Challenges regardingsamplepreparationandextractiontechniquesaredescribed.Differentapproachesusingthin-lay-erchromatographyarediscussed,andespeciallythepossibilityofsimultaneousdeterminationofparent,metaboliteandinterferingdrugsinurinesamples,aswellastheidentificationofmarkersforcombinedconsumptionofcocaineandalcohol.

toxicology,comparisonofmethods,review 1,22

10500� A.ZEB*,M.MURKOVIC(*Institute forBiochemistry,GrazUniversityofTechnology,Graz,Austria;[email protected]):Thin-LayerChromatographicanalysisofcarotenoidsinplantand animal samples. J.PlanarChromatogr. 2�, 94-10� (2010)This reviewdescribes availabledataonanalysisofcarotenoidsbyTLC.Petroleumether,acetone,andhexanearethemajormo-bilephasesusedforTLC.Thistechniquewasfoundtohavethepotentialtobethefirstchoiceforanalysisofcarotenoidsinbiologicalsamples.Theusesofother,orthogonalchromatographicmethods,forexampleHPLC,MS,scanningdensitometry,andimageanalysiswithTLCcanen-ablepreciseanalysisofcarotenoids.Thereviewconsistsofthefollowingparts:1.Introduction;1.1Functionofcarotenoids;1.2Occurrenceofcarotenoids;2.Analysisofcarotenoids;2.1Sam-pling;2.2Samplepreparation;2.�Extractionofcarotenoids;2.4Saponification;2.5Chromato-graphicanalysis;�.Thin-layerchromatographicanalysisofcarotenoids;�.1TLCofcarotenoidsfrommicrobialandanimalsources;�.2TLCofcarotenoidsfromplantsources;�.�Normal-pha-seTLCanalysisofcarotenoids;�.4Reversed-phaseTLCanalysisofcarotenoids;�.5TLCana-lysisofcarotenoidswithscanningdensitometry;4.AdvantagesofTLCincarotenoidanalysis;5.Conclusionandfuturestudies.1�4References.

foodanalysis,review 1,�0b

2. Fundamentals, theory and general

105004 Rodica Domnica BRICIU*, Agata KOT-WASIK, A. WASIK, J. NAMIESNIK, C. SARBU(*Babes-BolyaiUniversity,FacultyofChemistryandChemicalEngineering,AranyJanosStr.,No11,400028ClujNapoca,Romania):Thelipophilicityofartificialandnaturalsweetenersesti-matedbyreversed-phasethin-layerchromatographyandcomputedbyvariousmethods.J.Chro-matogr.A1217(2�),�702-�706(2010).Evaluationof thechromatographicbehaviorof someartificial and natural sweeteners by HPTLC on RP18, RP18W, RP8, cyano and amino phaseswithmixturesofacetonitrile-waterindifferentvolumeproportions.ThelipophilicityisgiventhroughchromatographicdescriptorssuchasRM0,meanofRM(mRM),andscoresofRMva-luescorrespondingtothefirstprincipalcomponent(PC1/RM).Inaddition,scoresandloadingsresultingfromcovariancematrixofretentiondataprovidenewinformationaboutsimilarityanddifferencesofinvestigatedcompoundsandbetweenthestationaryphaseandthemobilephases.Theexperimentallipophilicityindicesestimatedfromretentiondatawerecorrelatedwithcom-

CAMAGBIBLIOGRAPHYSERVICE No.1054

putedvalues,viacomputersoftwareandinternetmodule.Resultswereinaccordanceatahighlysignificantstatisticallevel.

qualitycontrolfoodanalysis,HPTLC 2c

105005 K. FERENCZI-FODOR, B. RENGER*, Z. VÉGH (*Vetter Pharma-Fertigung GmbH & Co.KG,Schuetzenstrasse87,88212Ravensburg,Germany;[email protected]):Thefrustratedreviewer-recurrantfailuresinmanuscriptsdescribingvalidationofquantitativeTLC/HPTLCproceduresforanalysisofpharmaceuticals.J.PlanarChromatogr.2�,17�-179(2010).Manymanuscriptsandalreadypublishedarticlesonanalyticalprocedurestobeusedinpharma-ceuticalqualitycontrolarecharacterizedbyseveraltypicalmethodologicalfailuresandmiscon-ceptions.Theautorspresentacollectionoftypicalfailures,misconceptions,andmisleadingdatafromarticlespublishedoverthelasttwoyearsinsevenwell-knownchromatographicpublicationsandprovideatthesametimealistofreferencesdescribingoptimumapproachestovalidationofspecificTLC/HPTLCprocedures.Inparticular,methodspecificity,linearity,accuracy,andpreci-sionveryoftenarenotdeterminedproperlyandinaccordancewithbestpractise.

qualitycontrol,HPTLC,quantitativeanalysis,validationofmethods 2f

105006 T.HANAI(HealthResearchFoundation,InstitutPasteur5F,Sakyo-ku,Kyoto606-8225,Japan;[email protected]):Quantitative insilicoanalysisof retention innormalphase liquidchro-matography.J.Liq.Chromatogr.Relat.Technol.��,297-�04(2010).Retentioninnormalphaseliquidchromatographicwasquantitativelyanalyzedinsilicousingassamplessleepingmedicinesinacidic,basic,andneutralorganicsolventmixtures.Hydrogenbondingisthemajorcontributionintheretentionthatwassupportedbyahighcorrelationcoefficientbetweenlogkandhydrogenbondingenergyvaluescalculatedusingamolecuarmechanicsprogram.TLConsilicagelwithchloroform-acetone9:1,benzene-aceticacid9:1anddioxane-benzene-aqueousammoniumhydroxide20:75:4.

2c

3. General techniques

105007 V.BEREZKIN*,SvetlanaKHREBTOVA,NataliaKULAKOVA(*Topchiev InstituteofPetro-chemicalSynthesis,RussianAcademyofSciences,Leninskypr.29,Moscow,119991,Russia):Four-dimensionalTLC on plates with open and closed adsorbent layers. Chromatographia 71(9-10),907-911(2010).Descriptionoffour-stageTLC,anewversionofmultidimensional(mul-ti-stage)planarchromatography.Inthefirststageamulticomponentmixtureisfractionatedintothreegroupsofchromatographiczones(fractions)byTLCwheretheadsorbentisdepositedonanaluminumfoil.Inthesecondstage,theTLCplateisdriedandcutbyscissorsintothreenar-rowstripscontainingtheseparatedfractions.Inthenextstagestheisolatedfractionsoneachofthese strips („daughter“ plates) are separated into individual components with an appropriatemobilephaseandbyusinga solventflowperpendicular to theflowof themobilephase.Thepracticalapplicationofthemethodisshownusingasanexampletheseparationofamixtureofdyes.Themethodcanbeexpandedforseparationofmixturescontainingfourormorefractions.Thisfour-dimensionalversionofmultidimensionalTLCwascarriedoutonplateswithbothopen(conventional)andclosedadsorbentlayers.

�d

105008 J.E.CLARK*,SusanOLESIK(*TheOhioStateUniversity,DepartmentofChemistry,120West18thAve,Columbus,OH4�210,USA):Electrospunglassycarbonultra-thinlayerchromatogra-phydevices. J.Chromatogr.A1217 (2�), 4655-4662 (2010).Development and applicationofelectrospunglassycarbonnanofibers forultra-thin layerchromatography (UTLC).ThecarbonnanofiberstationaryphasewascreatedthroughelectrospinningandpyrolysisofSU-82100pho-toresist,whichresultedinglassycarbonnanofiberswithdiametersof200-�50nmthatforma


matstructurewithathicknessof15µm.ThechromatographicpropertiesofUTLCdevicespro-ducedfrompyrolyzedSU-8heatedtotemperaturesof600,800,and1000°Cwereinvestigated.ByuseofRaman spectroscopy and scanning electronmicroscopy thephysical andmolecularstructureofthenanofibersateachtemperaturewasdetermined.ThecarbonUTLCdevicesweresuitablefortheanalysisvariousdyemixturesandalsoallowedseparationofthreeFITC-labeledessentialaminoacids (lysine, threonine,phenylalanine).TheelectrospunglassycarbonUTLCplatesshowedgoodretentionproperties,platenumbervaluesabove10000,andphysicalandche-micalrobustnessforarangeofmobilephases.

doping,HPTLC,qualitativeidentification,quantitativeanalysis �b

105009 P.PLOCHARZ*,AnnaKLIMEK-TUREK,T.H.DZIDO(*DepartmentofPhysicalChemistry,Medical University of Lublin, Staszica 6, 20-081 Lublin, Poland): Pressurized planar electro-chromatography, high-performance thin-layer chromatography and high-performance liquidchromatography-Comparisonofperformance.J.Chromatogr.A1217(2�),4868-4872(2010).Comparisonofthekineticperformance,measuredbyplateheight,ofhigh-performancethin-lay-erchromatography(HPTLC),high-performanceliquidchromatography(HPLC)andpressurizedplanarelectrochromatography(PPEC).HPTLConRP18Wwithmobilephasescomposedoface-tonitrile-buffer.TheHPLCcolumnwaspackedwiththesameadsorbent,whichwasscrappedfromanRP18WHPTLCplate.AnadditionalHPLCcolumnwaspackedwithC18-typesilicaba-sed(LiChrosorbRP18)adsorbentof5µmparticlediameter.TheplateheightofbothHPLCandPPECsystemsdependsontheflowvelocityandthemigrationdistanceofthemobilephase.Astestsolutionprednisolonesuccinatewasused.ThebestperformancewasobtainedbythePPECsystem.Theseparationefficiencyofthesystemswasinvestigatedandconfirmedbyuseofatestcomponentmixturecomposedofsixhormones.

qualitycontrol,HPTLC,comparisonofmethods,pressurizedplanarelectrochromatography �

4. Special techniques

105010 P. HOFFMANN, M. HULSEWIG, S. DUVAR, H. ZIEHR, M. MORMANN, J. PETER, A.FRIEDRICH,H.KARCH,J.MUTHING*(*InstituteofHygieneandInterdisciplinaryCenterforClinicalResearch,UniversityofMunster,Munster,Germany,[email protected]):OnthestructuraldiversityofShigatoxinglycosphingolipidreceptorsinlymphoidandmyeloidcellsde-terminedbynanoelectrosprayionizationtandemmassspectrometry.RapidCommun.MassSpec-trom.24,2295-2�04(2010).HPTLCofglycosphingolipids(GSLs)onsilicagelwithchloroform-methanol-water120:70:17.TheplatewasoverlaidwithShigatoxin(Stx),andthemicrobesweredetectedwithprimaryanti-Stxandappropriatealkalinephosphataselabeledsecondaryantibo-dies.Boundantibodieswerevisualizedbycolordevelopmentusing0.05%5-bromo-4-chloro-�-indolylphosphatep-toluidinesalt inglycinebuffer.TheGSLswereextractedfromtheplateand analyzed by electrospray ionization mass spectrometry (ESI-MS). Using the combinationofaTLCoverlayassayandnanoESI-QTOF-MS,thestructuralcharacterizationofthefunctionalStx1receptorsofRajiandTHP-1cellsisreported.

pharmaceuticalresearch,HPTLC 4e

105011 A. MUELLER, S. WEISS, W. SCHULZ*, W. SEITZ, R. ALBERT, W. RUCK, W. WEBER(*Zweckverband Landeswasserversorgung, Betriebs- und Forschungslaboratorium, Am Spit-zigen Berg 1, 89129 Langenau, Germany, [email protected]): Combination of differentliquidchromatography/massspectrometry technologies for the identificationof transformationproductsofrhodamineBingroundwater.RapidCommun.MassSpectrom.24,659-666(2010).HPTLCof rhodamineBandfivede-ethylated transformationproducts (N,N,N‘-tryethylrhoda-mine (1), N,N‘-dyethylrhodamine (2), N,N-dyethylrhodamine (�), N-ethylrhodamine (4), andrhodamine(5))ingroundwateronsilicagelbyautomatedmultipledevelopmentwitha2�-stepgradientbasedonmethanol(withtheadditionofformicacid)anddichloromethane.Thedrying


timeaftereachstepwas2min.Fordetectionbybioluminescencetheplatewasdippedintoasus-pensionofVibriofischerifor2sataspeedof�cm/s.ThehRfwere72,66,60,5�,48,and�6forcompounds(1)-(5).Combinationofdifferentseparationanddetectiontechniquesenabledafastandeffectivescreeningofthegroundwatersample.

environmental,HPTLC,qualitativeidentification,postchromatographicderivatization,AMD 4e

105012 A.MUSKEN,J.SOUADY,K.DREISEWERD,W.ZHANG,U.DISTLER,J.PETER,H.MIL-LER,H.KARCH,J.MUTHING*(*InstituteofHygieneandInterdisciplinaryCenterforClini-calResearch,UniversityofMunster,Munster,Germany, [email protected]):Applicationofthin-layerchromatography/infraredmatrix-assistedlaserdesorption/ionizationorthogonaltime-of-flightmassspectrometrytostructuralanalysisofbacteria-bindingglycosphingolipidsselectedbyaffinitydetection.RapidCommun.MassSpectrom.24,10�2-10�8(2010).HPTLCofglycos-phingolipids(GSLs)onsilicagelwithchloroform-methanol-water120:70:17.TheplatewasoverlaidwithGSL-specificbacteria,andthemicrobesweredetectedwithprimaryantibodiesandappropriate alkaline phosphatase labeled secondary antibodies, and by in situ MS analysis ofbacteria-specificGSLreceptors.Thethin-layerchromatographyinfraredmatrix-assistedlaserde-sorption/ionization orthogonal time-of-flight mass spectrometry (TLC/IR-MALDI-o-TOF-MS)methodrepresentsoneof themostpowerfulapproachesfor thedetectionofGSLreceptorsofmicroorganisms.

pharmaceuticalresearch,HPTLC 4e

10501� K.NIMPTSCH,R.SUESS,T.RIEMERA,A.NIMPTSCH,M.SCHNABELRAUCH,J.SCHIL-LER(*UniversityofLeipzig,MedicalFaculty,InstituteofMedicalPhysicsandBiophysics,Här-telstr.16-18,04107Leipzig,Germany):Differentlycomplexoligosaccharidescanbeeasilyiden-tifiedbymatrix-assistedlaserdesorptionandionizationtime-of-flightmassspectrometrydirectlyfromastandardthin-layerchromatographyplate.J.Chromatogr.A1217(2�),�711-�715(2010).Oligosaccharides(derivedfromdextran,alginate,hyaluronanandchondroitinsulfate)werecha-racterizedbymatrix-assistedlaserdesorptionandionizationtime-of-flight(MALDI-TOF)massspectrometry (MS) directly on a normal phaseTLC plate.The applied oligosaccharides wereeithercommerciallyavailableorobtainedfromthepolysaccharidesbyHCl-inducedhydrolysis.TLCwasfollowedbyMALDI-TOFMSsubsequenttomatrixdeposition.Thehighqualitymassspectraobtainedallowforunequivocalassignments.Thehighcontentofformicacidinthesol-ventdoesnotcausemajorproblemsbutisresponsibleforthepartialformylationoftheanalyteandaminorN-acetyllossfromhyaluronanandchondroitinsulfate.

qualitativeidentification,HPTLC,oligosaccharides 4e

105049 W.SCHWACKetal.,seesection�0

6. Alcohols

105014 AnnaNIESTROJ(SilesianUniversity,InstituteofChemistry,9SzkolnaStreet,40-006Katowice,Poland;[email protected]):Comparisonofmethodsforcalculationofthepartitioncoefficientsofselectedaliphaticcompounds.J.PlanarChromatogr.2�,198-200(2010).PropositionofnewmethodsforcalculationofthepartitioncoefficientsofaliphaticcompoundsfromexperimentalRfvaluesandthenumericalvaluesofselectedtopologicalindexes.Theexperimentalpartitioncoefficient(logPexp)ofcetylalcoholwasdeterminedforthen-octanol-watersystem.Numericalvaluesobtainedwerecomparedwiththeoreticalvaluesfromadatabase(AlogPs,AC_logP,AB/LogP,ALOGP,milogP,andXLOGP2).HPTLCofcetylalcohol,stearylalcohol,palmiticacid,stearicacid,alpha-hydroxypalmiticacid,and12-hydroxystearicacidonRP18withmethanolandwithmethanol-water19:1inahorizontalchamberatroomtemperature.Detectionaftervisuali-zationiniodinevapor.


comparisonofmethods,qualitativeidentification 6,11a

7. Phenols

105015 T. HOFMANN*,T. RÉTFALVI, L.ALBERT, P. NIEMZ (*University of West Hungary, De-partment for Chemistry,Ady Endre u. 5, 9400 Sopron, Hungary; [email protected]):High-performancethin-layerchromatographicassessmentofthermallymodifiedwood.J.PlanarChromatogr.2�,227-229(2010).HPTLCofphenoliccompounds(syringicacid,vanillicacid,pyrocatechin, thymol,sinapicacid,2,6-dimethoxyphenol,resorcin,guaiacol)onsilicagelwithdiisopropylether-formicacid9:1inanunsaturatedtwintroughchamber.OPLCofsugars(suc-rose,galactose,glucose,mannose,fructose,arabinose,xylose,ribose,rhamnose,stachyose,raf-finose,maltose)onsilicagelwithacetonitrile-water17:�.DetectionofphenoliccompoundsbysprayingwithFolin-Ciacalteaureagentandofsugarsbysprayingwithaniline-diphenylaminereagent.

agricultural,HPTLC,qualitativeidentification 7

105016 SayyadaKHATOON*,H.SINGH,K.SINGH,A.K.GOEL(*PharmacognosyandEthnophar-macologyDivision,CouncilforScientificandIndustrialResearch,NationalBotanicalResearchInstitute, Rana Pratp Marg, Lucknow-226001, India; [email protected]):TLC evaluationandquantificationofphenoliccompoundsindifferentpartsofDendrophthoefalcata(Linn.f.)Etting.J.PlanarChromatogr.2�,104-107(2010).TLCof(+)-catechin,ellagicacid,quercetin,and ferulic acid on silica gel with toluene - ethyl acetate - fomic acid 6:4:1 in a twin-troughchamberpreviouslysaturatedfor�0minat25°C.Othermobilephasecompositionswereethylacetate-methanol-water10:1:1,n-butanol-aceticacid-water4:1:5(upperlayer),n-butanol-ethanol-water20:5:11,n-butanol-aceticacid-water6:2:1,toluene-ethylacetate-formicacid5:5:1 and7:�:1, and toluene - ethyl acetate -methanol - formic acid140:60:1:1.Quantitativedeterminationbyabsorbancemeasurementat�20nm.Detectionbydippinginanisaldehydere-agent,followedbydryingandheatingat110°Cfor5min.Characteristicbandsof(+)-catechin,ellagicacid,quercetin,andferulicacidwereobservedathRf �5,41,6�and66, respectively.Precision(CV,n=7)was0.�9,0.44,0.55and0.16%andrepeatability(CV,n=7)0.65,0.95,0.�7and0.18%,respectively.LODwas54,�40,48and78ng/band,andLOQ4.1,6.5,1.6and1.5µg/band,respectively.Linearregressionwas0.999�(100-500ng/band),0.988�(1000-5000ng/band),0.9989(100-500ng/band)and0.9987(100-500ng/band),respectively.

Noteoftheeditor:ThereportedLOQareunusallyhighcomparedtotheLOD.ThereportedCV(standarddeviation

dividedbymean)alsoarisesquestions.

herbal,traditionalmedicine,densitometry 7

105017 M.M.PANDEY,S.RASTOGI,A.K.S.RAWAT* (*PharmacognosyandEthnopharmacologyDivision,NationalBotanicalResearchInstitute,Lucknow-226001,India;[email protected]): Optimization of an HPTLC method for separation and identification of phenolic com-pounds.J.PlanarChromatogr.2�,108-111(2010).HPTLCofgallicacid,ferulicacid,syringicacid,catechin,protocatechuicacidandvanillinonsilicagelwithtoluene-ethylacetate-formicacid8:2:1inatwin-troughchamberpreviouslysaturatedfor15min.Quantitativedeterminationby absorbance measurement at 280 nm.Accuracy was between 96.� and 90.7 %, repeatabili-tybetween0.65and0.9�%,inter-dayprecisionbetween0.80and0.90%,intra-dayprecisionbetween0.72and0.95%,andprecisionbetween0.87and0.92%.LODandLOQwereabout100and400ng/band, respectively.StartingatLOQ, thecorrelationcoefficientswerebetween0.988and0.997.

herbal,traditionalmedicine,HPTLC 7

8. Substances containing heterocyclic oxygen

105018 V.GLAVNIK,B,SIMONOVSKA,IrenaVOVK*(*NationalInstituteofChemistry,Laboratory


forFoodChemistry,Hajdrihova19,SI-1001Ljubljana,Slovenia;[email protected]):ComparisonofTLCandHPLCmethodsusedforanalysisof(-)-epicatechinanditsdimerprocyanidinB2inchocolate. J.PlanarChromatogr.2�,2�0-2�2 (2010).HPTLCof (+)-catechin, (-)-epicatechin,(-)-epigallocatechin,(-)-epigallocatechingallate,(-)-epicatechingallate,procyanidinB2,procya-nidinA2,andmethylxanthines(theobromineandcaffein)oncellulosewithn-propanol-water-aceticacid20:80:1 inahorizontalchamber.Detectionbydipping for1s into4-dimethylami-nocinnamaldehydedetectionreagent.Quantitativedeterminationbyabsorbancemeasurementat655nm.BydensitometryLODfor(-)-epicatechinandprocyanidinB2was0.2and2ng/zone,respectively;LOQwas0.4ngand4ng/zone,respectively.Theselimitswerelowerbyafactor50for(-)-epicatechinandbyafactorof10forprocyanidinB2thanthoseobtainedbyHPLC.TheTLCmethodgaveamoreaccurateresultforthe(-)-epicatechincontentofbakingchocolatethantheHPLCmethodwhichwasalsomoretime-consuming.

foodanalysis,comparisonofmethods,HPTLC,densitometry,quantitativeanalysis 8a

105019 J.SUN(SunJia),Y.YUE*(YueYongde),F.TANG(TangFeng),X.GUO(GuoXuefeng)(*In-ternationalCentreforBambooandRattan,No.8FutongDongdajie),Wangjing,ChaoyangDis-trict,Beijing100102,Cina;[email protected]):SimultaneousHPTLCanalysisofflavonoidsintheleavesofthreedifferentspeciesofbamboo.J.PlanarChromatogr.2�,40-45(2010).HPTLCof flavonoids (vitexin, isovitexin, orientin, isoorientin, rutin) on polyamide at 2� +/- 2°C and40%relativehumiditywiththethree-componentmobilephaseA-B-C��:67:8,whereAisdo-decylsulfate-n-butanol-n-heptane147:158:28,Biswater,andCisformicacid.Thesolventmentionedgavethebestresolutionofvitexin(hRf61),isovitexin(hRf21),orientin(hRf28),isoorientin(hRf�4),andrutin(hRf52).Detectionbysprayingwith1%aluminiumtrichlorideinethanolfollowedbywaitingforapproximately1h.Quantitativedeterminationbyfluorescencemeasurementat�66nm.Precisionwasfoundtobe0.98,0.91,1.02,1.04and0.87%forisovite-xin, rutin,orientin, isoorientin,andvitexin, respectively.Average recoveries (at threedifferentconcentrations)werebetween98.9and100.6%fromP.pubescens,P.glaucaandP.yixingensis.

HPTLC,densitometry,quantitativeanalysis,qualitativeidentification 8a

9. Oxo compounds, ethers and epoxides

105020 P.DEEPIKA,K.N.SABHARWAL,N.SIVARAMAN,T.G.SRINIVASAN*,P.R.VASUDEVARAO(*FuelChemistryDivision,ChemistryGroup,IndiraGandhiCentreforAtomicResearch,Kalpakkam,TamilNadu60�102,India;[email protected]):Developmentofachromatographicprocedure for thepurification of 1,2-diketone. J.Liq.Chromatogr.Relat.Technol. ��, 97-108(2010).TLCofoctane-4,5-dioneonsilicagelwithhexane,dichloromethane,andhexane-di-chloromethane1:1.Detectionbyplacingtheplateinaiodinechamber.

qualitativeidentification 9

10. Carbohydrates

105021 J.GEISSER*,EvamariaKRATZ(*ChemicalandVeterinaryInvestigationLaboratory(CVUA),WeissenburgerStr.�,76187Karlsruhe,Germany,[email protected]):Determina-tionofaloeveragelincosmetics.CBS104,1�-15(2010).HPTLCofaloeverose,glucoseandgalactoseinaloeveraproductsonsilicagel(impregnatedwithNaH2PO4)withacetone- iso-propanol-0.1Mformicacid2:2:1(doubledevelopmentforproductswithhighglucosecontent).Detectionbyimmersionin4-aminobenzoicacidreagent(1g4-aminobenzoicacidin�6mLace-ticacid,with40mLwater,2mL85%phosphoricacidand120mLacetone),followedbyhea-tingat100°Cfor10min.Densitometricevaluationbyfluorescencemeasurementat�66nm.Thelimitofquantificationwasapprox.�%inaloeveraproducts.RSDforpolynomialcalibrationis2.1%formannoseintherangeof8-80ng/bandand4.0%usingmatrixcalibration.

herbal,HPTLC 10b


105022 P.MANDAL,A.MISRA*(*DivisionofMolecularMedicine,BoseInstitute,A.J.C.BoseBirthCentenaryCampus,Kolkata,India,[email protected]):Concisesynthesisofthepentasac-charideO-antigencorresponding to theShiga toxinproducingEscherichiacoliO171.Bioorg.Chem.�8,56-61(2010).TLCof4-methoxyphenylglycosideonsilicagelwithacetonitrile-me-thanol-water4:2:1.Detectionbysprayingwithwarm2%cericsulphatein2Nsulfuricacid.ThehRfof4-methoxyphenylglycosidewas�0.

pharmaceuticalresearch,qualitativeidentification 10b

10502� K.SONG,J.SHIM,J.PARK,S.KIM,Y.KIM,W.BOOS,K.PARK*(*DepartmentofBiology,UniversityofIncheon,Incheon,RepublicofKorea,[email protected]):Transglycosylationpropertiesofmaltodextringlucosidase(MaIZ)fromEscherichiacolianditsapplicationforsyn-thesisofanigerose-containingoligosaccharide.Biochem.Biophys.Res.Commun.�97,87-92(2010).PreparativeTLCofthehydrolysisreactionofmaltodextringlycosidaseonvarioussubs-trates(maltotriose,alpha-,beta-,gamma-cyclodextrinsandcycloamylose)onsilicagelwithethylacetate-methanol-aceticacid-water12:�:�:1.ThestructureoftheproductswasdeterminedbyMALDI-TOF/MS,1�CNMR,andenzymaticanalysis.

pharmaceuticalresearch,preparativeTLC 10b

105024 C.YANG(YangCheng),J.GUAN(GuanJia),J.-S.ZHANG(ZhangJiang-sheng),S.-P.LI*(LiShao-ping)(*InstituteofChineseMedicalSciences,UniversityofMacau,MacauSAR,China;[email protected]):UseofHPTLCtodifferentiateamongthecrudepolysaccharidesinsixtraditionalchinesemedicines.J.PlanarChromatogr.2�,46-49(2010).HPTLCofpolysac-charides (with galactose, glucose, mannose, arabinose, ribose, xylose, rhamnose, galacturonicacid,andglucuronicacidasstandards)beforeandafterhydrolysisonsilicagelwithchloroform-n-butanol - methanol - water - acetic acid 9:25:10:�:� with chamber saturation for �0 min atroomtemperature.Detectionbysprayingwithaniline-diphenylamine-phosphoricacidreagentandheatingat1�0°Cfor10min.Quantitativedeterminationbyabsorbancemeasurementat�80nm.Ninhydrinreagentwasusedfordetectionofothercompounds.

pharmaceuticalresearch,herbal,traditionalmedicine,HPTLC,densitometry 10b

11. Organic acids and lipids

105025 P.KUSHWALI*,SheejaEDWIN,K.VARSHNEY,E.JARALD,S.AHMAD,A.DAUA(*TI-FACOREinGreenPharma,B.R.NahataCollegeofPharma,Mhow-NeemuchRd.,Mandsaur(M.P.),India):Estimationofcurcuminand�-acetyl-11-keto-a-boswellicacidinamarketedher-balproduct rheumaxusingHPTLC. InternationalSeminaronHerbalDrugResearch,PN-028(2009).HPTLCofcurcuminand�-acetyl-11-keto-a-boswellicacidintheherbalproductRheu-max (containsCurcuma longa,Boswellia serrata,Tinospora cordifolia andVitexnegundo)onsilicagelwithchloroform-methanol�7:�forcurcuminandn-hexane-ethylacetate1:1fortheacid.Quantitativedeterminationbyabsorbancemeasurementat4�0nmforcurcuminand254nmfortheacid.Themethodwaslinearintherangeof100-500ng/bandforcurcuminand1500-4000ng/bandfortheacid.

traditionalmedicine,quantitativeanalysis,HPTLC 11a

105014 AnnaNIESTROJ,seesection6

105026 K.RIZWANBASHA*,M.SHANMUKHA,K.RAMRAO,N.MUNJUNATHA,V.SENTHIL,M.SAMANTA(*JSSCollegeofPharmacy,Dept.ofPharmaceutics,Oaty,India):Designanddevelopmentof triphala fastdispersable tablets and its characterization. InternationalSeminaronHerbalDrugResearch,PN-064(2009).HPTLCofgallicacidasmarkercompoundintriphala


fastdispersabletabletsonsilicagelwithethylacetate-toluene-methanol-glacialaceticacid75:20:�:2.ResultsfromHPLCanalysiswerecomparable.Themethodwassuitableforroutinequalitycontrolofdispersabletabletsformulation.

herbal,densitometry,HPTLC 11a

105027 KarinROTHENBUEHLER,E.REICH*,M.HAMBURGER (*CAMAGLaboratory,Sonnen-mattstr.11,41�2Muttenz,Switzerland):ValidatedHPTLCmethodforskinlipids.CBS1045-6(2010).HPTLCofskinlipids(squalene,triolein,palmiticacid,1,2-dipalmitoyl-sn-glycerol,stea-rylpalmitate,cholesterylpalmitate,andcholesterol)onsilicagel(prewashedwithmethanol)firstwithtoluenetoadevelopingdistanceof80mm,thenwithn-hexane-t-butylmethylether-aceticacid80:20:1to45mminatwin-troughchambersaturatedfor20min.Detectionbydippingincopper(II)sulfatereagent followedbyheatingfor�0minat140°C.Densitometricabsorbancemeasurementat�50nm.Stabilityofstandardsandsamplesduring�h insolutionandon theplatewasgood.ThedifferencesofthehRfvaluesof7compoundson�plateswasgood(<�).TheprecisionofmeasurementwaslowestonHPTLCLiChrosphersilicagelplates(RSDof<5%,n=9).Thelinearrangeforallbuttwocompoundswasbetween100and�50ng/bandandcorrela-tioncoefficientswere>0.9975.

HPTLC,pharmaceuticalresearch 11c

13. Steroids

105028 K.BOBER(FacultyofPharmacy,DepartmentofAnalyticalChemistry,MedicalUniversityofSilesia,4JagiellonskiStreet,PL-41-200,Sosnowiec,Poland;[email protected]):Densitometryapplicationforevaluationofthevisualizingagentsfordehydroepiandrosterone.J.Liq.Chroma-togr.Relat.Technol.�1,267�-2685(2008).TLCofdehydroepiandrosteroneonsilicagelwithchloroform-acetone17:�withchambersaturationfor�0min.Detectionbydippinginto0.05%aqueoussolutionsofvisualizingagents(methyleneviolet,gentianviolet,janusblue,methyleneblue,malachitegreen,rhodamineB)andmethanolicchloramineT/methanolicsulfuricacidso-lution.Quantitativedeterminationbydensitometryat200nm(beforederivatizationandforme-thyleneviolet),657nm(gentianviolet),487nm(janusblue),488nm(malachitegreen),580nm(rhodamineB),and�61nm(chloramineT).LODwas0.29µg/5mL(withoutvisualizingagent,withchloramineTandwithrhodamineB),4.19µg/5mLwithmethyleneviolet,1.10µg/5mLwithgentianviolet,2.14µg/5mLwithjanusblue,and4.19µg/5mLwithmalachitegreen.

densitometry,quantitativeanalysis 1�a

15. Terpenes and other volatile plant ingredients

105029 B.QIN, J.EAGLES,F.MELLON,P.MYLONA,L.PEÑA,A.OSBOURN*(*SainsburyLa-boratory,NorwichNR47UH,UK,[email protected]):High throughput screeningofmutantsofoat thataredefectivein triterpenesythesis.Phytochemistry.71,1245-1252(2010).TLCofsqualene(1),2,�-oxidosqualene(2)andbeta-amyrin(�)intherootsofAvenastrigosaonsilicagelwithhexane-chloroform2�:2,hexane-chloroform1:1,andhexane-acetone4:1for(1),(2)and(�),respectively.Detectionbyexposuretoiodinevapor.QuantitativedeterminationbyGC-MS.

pharmaceuticalresearch,HPTLC,quantitativeanalysis,densitometry 15a

1050�0 A.VARMA,NeetaSHRIVASTAVA*(*B.V.PatelPharmaceuticalEducationandResearchDe-velopment(PERD)Centre,Sarkhej-GandhinagarHighway,Thaltej,Ahmedabad-�80054,Guja-rat,India;[email protected]):Microscaleprocedureforanalysisofandrogra-pholideinAndrographispaniculataleaves.J.PlanarChromatogr.2�,50-55(2010).HPTLCofandrographolideonsilicagelwithchloroform-methanol-ethylacetate12:�:2inatwin-troughchamber previously saturated for 15 min. Quantitative determination by absorbance measure-mentat22�nm.Therelativestandarddeviationofintra-dayandinter-dayanalysiswasintheran-


geof0.56-1.��%.Lineartiywasgivenbetween200-700ng/band;thecorrelationcoefficientwas0.9998andtheRSD0.97%.Thelimitsofdetectionandquantificationwere60and150ng/band.Averagerecoverywas98.8+-0.41%.

pharmaceuticalresearch,herbal,traditionalmedicine,HPTLC,densitometry,quantitativeanalysis 15a

17. Amines, amides and related nitrogen compounds

10506� R.DEEPAetal.,seesection�2

105087 B.MEHTAetal.,seesection�2

10511� JyotiSHRIVASTAVAetal.,seesection�2

18. Amino acids and peptides, chemical structure of proteins

1050�1 P.VENKATAKRISHNAN,E.NAKAYASU,I.ALMEIDA,R.MILLER*(*DepartmentofBiolo-gicalSciences,UniversityofTexasatElPaso,ElPaso,Texas,TheUnitedStatesofAmerica,[email protected]):Arginaseactivityinmitochondria-aninterferingfactorinnitricoxidesynthaseactivityassays.Biochem.Biophys.Res.Commun.�94,448-452(2010).HPTLCof[14C]-L-ar-ginineand[14C]-L-citrullineofanitricoxidesynthaseconversionassayinmitochondriaratliver,onsilicagelwithbutanol-aceticacid-water�:1:1.Detectionbydippinginasolutionof2%ninhydrin inacetone, followedbyheatingonaplateheater for1min.Aftervisualization, theplateswereexposedtoX-rayfilmfor4.5days.

pharmaceuticalresearch,HPTLC,quantitativeanalysis,autoradiography 18a

19. Proteins

1050�2 K.NISHIYAMA*,M.MAEDA,M.ABE,T.KANAMORI,K.SHIMAMOTO,S.KUSUMO-TO,T.UEDA,H.TOKUDA(*InstituteofMolecularandCellularBiosciences,TheUniversityofTokyo,Tokyo, Japan,[email protected]):Anovelcomplete reconstitution system formembraneintegrationofthesimplestmembraneprotein.Biochem.Biophys.Res.Commun.�94,7��-7�6(2010).TLCofliposomescontainingmembraneproteinintegrase(MPIase),onsilicagelwithethanol-chloroform-water7:�:4.Detectionbysprayingwithasolutionofanisalde-hyde-concentratedsulphuricacid-aceticacid�:4:1.ThehRfvalueofMPIasewas�5.

pharmaceuticalresearch,qualitativeidentification 19

21. Purines, pyrimidines, nucleic acids and their constituents

1050�� E. MINCSOVICS (OPLC-NIT Ltd,Andor Street 60, 1119 Budapest, Hungary, and CorvinusUniversity,FacultyofHorticulturalSciences,DepartmentofGeneticsandPlantBreeding,Bu-dapest,Hungary;[email protected]):Increaseddetectionsensitivityinfullyoff-lineOPLCbybilateralbandcompression.J.PlanarChromatogr.2�,190-192(2010).Aftersampleapplicationasbandsandfullyoff-lineOPLCseparationfollowedbydrying,theseparatedbandswerecompressedbilaterally,inparallel,perpendiculartothedirectionofdevelopment,byuseofastrongeluentandcapillarydrivenchromatography.Tointroducetheeluentforbandcompressi-onontothelayerasimplemanualtoolequippedwithparallelfoamstripswasconstructed.OPLCofablacktealeafextractandcaffeine,theophylline,andtheobromineonsilicagel(prewashedwithacetonitrile-water17:�)withtoluene-aceticacid�:2.Quantitativedeterminationbyden-sitometryat280nm.Afterbandcompression20ng/zonetheophyllinandtheobrominecouldbedetectedbydensitometryinthexanthinestandardmixtureataloadingof20µg/10mm.Thesecompoundswerenotvisibleintheoriginal,uncompressedchromatogram.

herbal,quantitativeanalysis,densitometry 21a


1050�4 K.VINODKUMAR*,T.VETRICHELVAN(*DeptofPharmaceuticalAnalysis,AdhiParasakthiCollege of Pharmacy,Tamil Nadu, India): Method development for risedronate sodium hemipentahydratebyUVspectroscopicmethodandhigh-performancethin-layerchromatography.IPAConvention,2010,RA-PO26.HPTLCofrisedronatesodiumonsilicagelwithwater-methanol-25%ammonia20:�:�.Densitometricevaluationat262nm.Themethodwaslinearintherangeof�-6ng/band,recoverywas98.6%.ResultswerecomparablewithHPLCresults.Theadvan-tageoftheHPTLCmethodisthesimultaneousanalysisofseveralsamples.

pharmaceuticalresearch,qualitycontrol,densitometry,comparisonofmethods,quantitativeanalysis,HPTLC 21a

22. Alkaloids

105002 MonikaJANICKAetal.,seesection1

1050�5 J.KENNEDY,J.WISEMAN*(*ProsoliaInc.,Indianapolis,IN46202,USA,[email protected]):DirectanalysisofSalviadivinoriumleavesforsalvinorinAbythinlayerchromatographyanddesorptionelectrosprayionizationmulti-stagetandemmassspectrometry.RapidCommun.MassSpectrom.24,1�05-1�11(2010).HPTLCofsalvinorinA(1),salvinorinC(2),divinatorinB(�),andsalvinorinB(4)intheleavesofSalviadivinoriumonsilicagelwithmethyltert-butylether-hexane�:1asmobilephase.Detectionbydesorptionelectrosprayionizationmulti-stagetandemmassspectrometry(TLC/DESI-MS).ThehRfwere49,64,85,and95forcompounds(1)-(4),respectively.TheuseofasimpleHPTLCprotocolincombinationwithDESI-MSallowedforimproveddetectionofdifferentspeciesofsalvinorinintheleafextracts.

toxicology,herbal,HPTLC,qualitativeidentification 22

23. Other substances containing heterocyclic nitrogen

1050�6 M. DOLOWY*,A. NIESTRÓJ (*Department ofAnalytical Chemistry, Faculty of Pharmacy,SilesianUniversityofMedicine,4JagiellonskaSt.,PL-42-200Sosnowiec,Poland;[email protected]):Densitometricdeterminationofursodeoxycholicacidinpharmaceuticalformulationsinformoftabletsandcapsules.J.Liq.Chromatogr.Relat.Technol.��,109-117(2010).TLCofur-sodeoxycholicacidonsilicagelwithn-hexane-ethylacetate-aceticacid22:22:5atroomtem-peraureinahorizontalchamber.ThehRfvaluewas48.Detectionbysprayingwith10%sulfuricacid and heating at 120 °C for 20 min. Quantitative determination by densitometric scanningat�60nm.LODandLOQwas0.014and0.041mg/spot,respectively.Thelinearityrangewas0.0�0-0.120mg/spot.Thecorrelationcoefficientrwas0.006�.Theaccuracyofthequantitativeanalysis of ursodeoxycholic acid in extracts from examined pharmaceutical formulations was95.�%and97.2%,andtheprecisionwas5.29and6.15%intabletsandcapsules,respectively.

qualitycontrol,densitometry,quantitativeanalysis 2�b

1050�7 AnnaPETRUCZYNIK(DepartmentofInorganicChemistry,MedicalUniversityofLublin,St-aszica6,20-081Lublin,Poland; [email protected]):Effectof chromatographicconditions on separation and system efficiency in HPTLC of selected quinoline standards oncyanopropylstationarypases.J.PlanarChromatogr.2�,56-64(2010).HPTLCofthirteenquino-lines(quinoline,isoquinoline,2,4-dihydroxyquinoline,8-hydroxyquinoline,5,6-benzoquinoline,2,2‘-diquinoline,8-methylquinoline,5-aminoquinoline,1-ethylquinoline,2-chlorquinoline,6-ni-troquinoline,2,6-dimethylquinoline,5-hydroxyquinoline)oncyanophasewithdifferentmobilephasespreparedfrommixturesofmethanol,acetonitrile,2-propanol, tetrahydrofuran,anddio-xanewithadditionofbufferatdifferentpH,ion-pairingreagents,orsilanolblockers.DetectionunderUVlightat254nmandbydensitometricabsorbancemeasurementat254nm.Improvedpeaksymmetrywasobservedformobilephasescontainingion-pairingreagent.Themostsym-metricalpeakswereobtainedbyuseofmobilephasecontainingdiethylamineassilanolblocker.

qualitativeidentification,HPTLC,densitometry 2�e


1050�8 T.M.SHAH*,S.S.SAVLE,M.T.CHHABRIA,I.S.RATHOD,C.J.SHISHOO,P.S.BRAHM-KSHATRIYA (*Dept. ofPharmaceuticalChem.&Dept. ofQ.A.,L.M.CollegeofPharma-cy,Navranga,Ahmedabad�80009,India):PharmacokineticstudyofanovelantihyperlipidemicagentLM-1�765-Aprodrug.Ind.J.Pharma.Science7161,644-650(2009).HPTLCofthenovelantihyperlipidemicagentLM-1�765inrabbitplasmaonsilicagel(pre-washedwithmethanol)withbenzene-methanol4:1inasaturatedchamberat25°C.Densitometricevaluationat�14nm.Themethodwaslinearintherangeof12.5-100ng/band.AlsoHPTLCofLM-1�765-C,ametaboliteoftheparentcompoundLM-1�765,onsilicagelwithtoluene-methanol2�:2.Densi-tometricevaluationat274nm.Detailsareprovidedonthecompleteextractionprocedurefortheextractionoftheparentcompoundanditsmetabolitefromplasma.

pharmaceuticalresearch,clinicalroutineanalysis,HPTLC,densitometry,quantitativeanalysis 2�b

24. Organic sulfur compounds

1050�9 K.PLUTA*,B.MORAK-MLODAWSKA,M.JELEN,R.KORLACKI(*DepartmentofOrganicChemistry,TheMedicalUniversityofSilesia,Jagiellonska4,41-200Sosnowiec,Poland;[email protected]):TLCseparationofisomericdiazinodithiinsanddiazinylsulfidesastheSmilesrearrangementproducts.J.Liq.Chromatogr.Relat.Technol.�1,�020-�0�1(2008).TLCoftwelfthioazinesonsilicagelwithchloroform-ethanol10:1andchloroform,andonaluminumoxidewithdichloromethaneandbenzene-chloroform1:1inachambersaturatedfor�0min.DetectionunderUV254and�66nm.Theretentionparametersweremeasuredandthencalculatedassepa-rationfactorsdeltaRf,RS,andalpha.ThehRfvalueswerecorrelatedwiththedipolemomentsofthioazinesandthesymmetryofdiazinodithiins.

qualitativeidentification 24

27. Vitamins and various growth regulators

105040 A.HOSU,ClaudiaCIMPOIU*,M.SANDRU,L.SESERMAN(*FacultyofChemistryandChe-micalEngineering,„Babes-Bolyai“University,11AranyJanos,400028Cluj-Napoca,Rumania;[email protected]): Determination of the antioxidant activity of juices by thin-layerchromatography.J.PlanarChromatogr.2�,14-17(2010).TLCofvitaminCofdifferentconcen-trations(0.45,0.50,0.55,0.60,and0.65mg/mL)andjuice(e.g.orangewithgrapefruit,orangewithappleandcarrot,„multi-fruit“)mixedwithamethanolicsolutionofDPPH(2,2-diphenyl-1-picrylhydrazyl)onsilicagel.Imagesofplateswereevaluatedusingaspecificcomputersoftware.Theequivalenceoftheresultsobtainedusingtheproceduredescribedwasdemonstratedbyap-plyingthetraditionalUV-visiblespectrophotometry.

Notefromeditor:Thetitleismisleading.ThisisnotTLCasthereisnochromatographyatall.Silicagelisusedas

carrierandnotasstationaryphaseforaseparation.

foodanalysis,qualitativeidentification 27

28. Antibiotics, Mycotoxins

105041 M.BROSZAT,C.WELLE,M.WOJNOWSKI,H.ERNST,B.SPANGENBERG*(*UniversityofOffenburg,InstituteofProcessEngineering,Badstrasse24,77652Offenburg,Germany;[email protected]):AversatilemethodforquantificationofaflatoxinsandochratoxinAindriedfigs.J.PlanarChromatogr.2�,19�-197(2010).HPTLCofochratoxinA,aflatoxinB1,G1,B2,andG2onsilicagelwitht-butylmethylether-water-methanol-cyclohexane48:1:2:1inanunsaturatedhorizontaldevelopingchamberandonRP18withmethanol-4%aqueouszincsulfatesolution-ethylmethylketone5:5:1.Afterdevelopmentthesilicagelplatewasdippedfor2sinsiliconeoil-hexane1:2whichenhancedaflatoxinfluorescencebyafactorof2andochra-toxinAfluorescencebyafactorof�-10.RP18platesweredevelopedtoadistanceof75mminan unsaturated vertical chamber.Averaged densitograms were obtained in the emission wave-length range from445 to485nm.SamplepretreatmentwasbymodifiedQuEChERS(Quick,Easy;Cheap,Effectice,Rugged,Safe)extractionwithtetahydrofuranoracetone.Linearitywas


intherangeof�to100pg/zoneforaflatoxinsB2andG2,10to�50pg/zoneforaflatoxinsB1andG1,and0.25to2.5ng/zoneforochratoxinA.LOQfortheaflatoxinswerebetween1�and�5pg/zone(equivalenttoto1.5and2.5ppb);forochratoxinAitwas970pg/zone(56ppb).

toxicology,HPTLC,densitometry,quantitativeanalysis 28b

105042 MicheleHOELTZ*,J.WELKE,I.NOLL(*InstitutodeCiênciaeTecnologiadeAlimentos,Uni-versidadeFederaldoRioGrandedoSul,Av.BentoGonçalves,9500,91570-901PortoAlegre-RS,Brasil,[email protected]):Photometricprocedureforquantitativeanalysisofaf-latoxinB1inpeanutsbythin-layerchromatographyusingchargecoupleddevicedetector.Quim.Nova��,4�-47(2010).HPTLCofaflatoxinB1inpeanutsonsilicagelwithchloroform-acetone99:1.Quantitativedeterminationbyabsorbancemeasurementat�66nm,usingaCCDcamerafollowedbyimageprocessingusingthesoftwareImageJ.Linearitywasbetween0.8and4.8ng/zone.Theintra-dayandinter-dayprecisionshadaRSDlowerthan5.2%.LODwas0.4ng/zonewhileLOQwas1.2µg/kg.Theaveragerecoverywas94.9%.Theproposedmethodisasimple,efficientandlowcosttoolforquantitativeanalysisofaflatoxinB1inpeanutsamples.

foodanalysis,HPTLC,quantitativeanalysis,densitometry 28b

10504� JulianeWELKE,M.HOELTZ,H.A.DOTTORI,I.B.NOLL(*InstitutodeCienciaeTecnologiadeAlimentos,UniversidadeFederaldoRioGrandedoSul,Av.BentoGoncalves,9500,91570-901,PortoAlegre,RS,Brasil;[email protected]):Rapid,simple,andeconomicalmethodforquantificationofochratoxinAinredwine.J.PlanarChromatogr.2�,116-118(2010).HPTLCofochratoxinAonsilicagelwithtoluene-ethylacetate-formicacid6:�:1inasaturatedcham-ber.Detectionbysprayingwithethanolicsodiumbicarbonatesolution(6gsodiumhydrogencar-bonate,100mLwater,20mLethanol);afterdrying,evaluationandvideodensitometryunder�66nm.ThelinearregressioncoefficientofthecalibrationforOTAintheconcentrationrange0.8to12ng/zonewas0.9992.Themeanrecoverywas92+-8.9%.Recoveryfromwinesamplesatlevelsof0.5,2,and5µg/Lwas84,90,and102%,respectively,andtherespectiverelativestan-darddeviationswere5.7,8,and7%.LODwas0.�2ng/spotandLOQ0.1µg/L.foodanalysis,HPTLC

quantitativeanalysis 28b

105044 JulianeWELKE*,MicheleHOELTZ,H.DOTTORI, I.NOLL(*InstitutodeCiênciaeTecno-logia deAlimentos,UniversidadeFederal doRioGrande doSul,Av.BentoGonçalves, 9500,91570-901PortoAlegre-RS,Brasil, [email protected]) :DeterminationofochratoxinAinwinebythin-layerchromatographyusingchargecoupleddevice.J.Braz.Chem.Soc.21,441-446(2010).HPTLCofochratoxinAinwineonsilicagelwithtoluene-ethylacetate-chlo-roform-formicacid6:�:1.Quantitativedeterminationbyabsorbancemeasurementat�66nm,usingaCCDcamerafollowedbyimagesprocessingusingthesoftwareImageJ.Linearitywasbetween0.8and�2µg/L.Theintra-dayandinter-dayprecisionshadaRSDlowerthan9.9%and11.5%,respectively.LODwas16ng/zonewhileLOQwas100ng/zone.Theproposedmethodisasimple,efficientandlowcosttoolforquantitativeanalysisofochratoxinAinwinesamples.

foodanalysis,HPTLC,quantitativeanalysis,densitometry 28b

29. Pesticides and other agrochemicals

105045 A.FITTLER*,B.KOCSIS,Z.MATUS,L.BOTZ(*PharmaceuticalInstituteandCentralPhar-macy,FacultyofGeneralMedicine,UniversityofPécs,Honvédu.�.,Pécs7624,Hungary;[email protected]):Asensitivemethodforthin-layerchromatographicdetectionofampho-tericinB.J.PlanarChromatogr.2�,18-22(2010).TLCofamphotericinBonsilicagelwithchlo-roform-methanol-boratebuffer(pH8.�)4:5:1inachamberpre-saturatedfor20min.DetectionunderUV�66nm.ThehRf of themaincomponentwas46,andof theminorcomponent�1.


Quantitativedeterminationbyabsorbancemeasurementat�85nm.DirectbioautographywithCandidaalbicansproved tobe themostsensitivemethod,withadetection limitof0.8ngperspot.Fordensitometricevaluationofplatesat�85nmtentimesmoresubstanceisrequired.

pharmaceuticalresearch,qualitycontrol,densitometry,quantitativeanalysis,comparisonofmethods 29e

105046 G.OROS,T.CSERHÁTI*(*ResearchInstituteofMaterials,andEnvironmentalChemistry,Che-mical Research Center, HungarianAcademy of Sciences, Budapest, Hungary; [email protected]):Relationshipbetweenthecalculatedphysicochemicalparametersandreversedphasethin-layerchromatographicretentionbehaviorofcarboxamidefungicidesandrelatedcompounds.J.Liq.Chromatogr.Relat.Technol.��,880-89�(2010).TLCof6carboxamidefungicidesand11phenylbenzamidederivativesonsilicagelandaluminiumoxideimpregnatedbyovernightprede-velopmentinn-hexane-paraffinoil19:1withmixturesofmethanol-water,acetonitrile-water,tetrahydrofurane-water,andacetone-waterwiththeconcentrationoforganicmodifiervaryinginstepsof5%.DetectionunderUVlight.TheRMOandbvaluesrelatedtothemolecularlipo-philicityandtothespecifichydrophobicsurfacearea(b)ofthesoluteswerecalculatedseparatelyfor eachRP-TLCsystemand for eachanalyte.Thecorrelationsbetween thephysicochemicalparametersmeasuredwerecalculatedbylinearregressionanalysis.

environmental,qualitativeidentification 29e

105047 T.TUZIMSKI(DepartmentofPhysicalChemistry,FacultyofPharmacy,MedicalUniversityofLublin,6StaszicaStreet,20-081Lublin,Poland;[email protected]):Newprocedureforanalysisofcomplexmixturesbyuseofmultidimensionalplanarchromatographyincombinationwithdiode-arrayscanningdensitometryandhigh-performance liquidchromatographycoupledwithdiode-arraydetection.J.PlanarChromatogr.2�,184-189(2010).MultidimensionalplanarchromatographyonmonolayerormultiphaseplatesandmodernfiberopticalTLCdensitometerscannerswithDADisespeciallyusefulforcorrectidentificationofcomponentsofdifficult,com-plicatedmixtures,e.g.pesticidesinplantextracts(afterpreliminaryclean-upandconcentrationby,e.g.,solid-phaseextraction).TLCofclofentezine[�,6-bis(2-chlorophenyl)-1,2,4,5-tetrazine]inHerbaThymionsilicagelinahorizontalchamberwithtetrahydrofuran-n-heptane�:7inthefirstdirection,thenwithethylacetate-n-heptane1:4intheseconddirection.Detectionintherangeof200to600nmwithaTLC-DADscanner.AlsoTLCofthymeherbextractsonsilicagelandonRP18plateswithtetrahydrofuran-n-heptane�:7inthefirstdirectionandwithmethanol-water7:�intheseconddirection.LODandLOQwere0.2�and0.70µg/band,respectively, inTLC-DADand0.�5and1.06µg/mL,respectively,inHPLC-DAD.Averagerecoveriesfromthespikedplantmaterialsampleswere80.1%and100.5%at2.5µg/gand95.1%at5µ/gmeasuredat202nm.

agricultural,herbal,densitometry,quantitativeanalysis 29

105048 R. ZAKRZEWSKI*, W. CIESIELSKI (*Department of Instrumental Analysis, University ofLódz,Pomorska16�,90-2�6Lódz,Poland;[email protected]):Thinlayerchromato-graphywithpost-chromatographiciodine-azidereactionforthiuramanalysisinfoodsamples.J.Liq.Chromatogr.Relat.Technol.�1,2657-2672(2008).TLCandHPTLCofthiuramonsilicagelwithmethanolordichloromethaneinasaturatedhorizontalchamberfor15min.Detectionbysprayingwithimprovedandnon-improvediodine-azide,iodine,andcopper(II)reagentsandbyevaluationunderUV254nm.ForderivatizationthedevelopedplatesweresprayedwithfreshlypreparedmixturesofsodiumazideandstarchsolutionadjustedtopH6.0(5mLof10%aqueoussodiumazidesolutionand12.5mLof2%aqueousstarchsolutionweremixedandadjustedtopH6.0with0.1mol/Lhydrochloricacidsolution;thesolutionwasdilutedto25mLwithwater)andexposedtoiodinevaporfor5s.Quantitativeanalysisbyscanningwithanofficescanner(PCscanner)andafterdetectionwithimprovediodine-azidereagentdensitometricmeasurementat


48�nm.ThehRfvalueofthiuramwas29.LODwere�and0.5pmolperspotusingaiodine-azidedetectionsysteminTLCandHPTLC,respectively.Linearitywasintherangeof2-8pmolperspot;thecorrelationcoefficientrwas0.9981.ResultsobtainedbyuseofaPCscannerwerecomparabletomeasurementsusingaTLCdensitometer.

foodanalysis,postchromatographicderivatization,HPTLC,quantitativeanalysis,densitometry 29e

30. Synthetic and natural dyes

105049 W.SCHWACK*,ElodiePELLISSIER(*UniversityofHohenheim,InstituteofFoodChemistry,Garbenstrasse28,70599Stuttgart,Germany,[email protected]):Determinationofunauthorisedfat-solubleazodyesinspicesbyHPTLC.CBS10�,1�-15(2009).HPTLCofazodyes(SudanI,II,III,IV,B,SudanorangeG,Sudanred7B,Parared)inspicesamplesoncaffe-ineimpregnatedsilicagelwithisohexane-methylethylketone5:1withchambersaturationfor10min.Densitometricabsorptionmeasurementat�90,415,500,525and550nm.Thelimitsofdetectionwereapprox.10mg/kg.Confirmationofsuspectedcompoundsinsamplesbycompari-sonofUVspectra.TLC-MSanalysisinpositiveESImodefurtherconfirmspositivefindings.

foodanalysis,HPTLC,quantitativeanalysis,densitometry,TLC-MS �0a,4e

105050 R. SKIBINSKI*, L. KOMSTA (*Department of Medicinal Chemistry, Medical University ofLublin,Jaczewskiego4,20-090Lublin,Poland,[email protected]):ValidationofNP-HPTLCandRP-HPTLCmethodswithvideodensitometricdetectionforanalysisofziprasidoneinpharmaceuticalformulations.J.PlanarChromatogr.2�,2�-27(2010).HPTLCofziprasidone(5-[2-[4-(1,2-benzothiazol-�-yl)piperazin-1-yl]ethyl]-6-chloro-1,�-dihydroindol-2-one)on silicagelwithhexane-dioxane-propylamine5:45:2upto9cm(undersaturatedconditions)andonRP8withtetrahydrofuran-phosphatebuffer(pH9.0)1:1upto4.5cm(underunsaturatedcondi-tions),bothinhorizontalchambers.Quantitativedeterminationbyvideodensitometryat254nm.Calibrationwaslinearintherange0.2-1.2and0.1-1.1µg/spotziprasidoneforNP-HPTLCandRP-HPTLC,respectively.Theintra-dayprecisionsfor0.4-1.2µg/spotonNP-HPTLCwas2.0to5.2%andonRP-HPTLC4.0to6.1%;therespectiveinter-dayprecisionforNP-HPTLCwas2.0to6.7%andforRP-HPTLC4.1to7.1%.LOD/LOQonNP-HPTLCwas0.0�/0.09µg/spot;usingRP-HPTLC,LOD/LOQwas0.02/0.06µg/spot.Thespecificityofthemethodswasconfir-medbycomparisonofhRfvalues(74+/-2inNP-HPTLCand�6+/-1inRP-HPTLC,n=12).qualitycontrol,HPTLC

densitometry �0a

105051 B.YUANGSOI*,O.JINTASATAPORN,P.TABTHIPWON,C.KAMEL(*DepartmentofAqua-culture, Faculty of Fisheries, Kasetsart University, Bangkok 10900, Thailand; [email protected]):Comparativepharmacokineticsafterfeedingfancycarp(Cyprinuscarpio)withdietscon-tainingcarotenoids fromnatural sources (tea (Camellia sinensis),mulberry (Morusalba), andcassava(Manihotesculenta)leaf).J.PlanarChromatogr.2�,219-224(2010).TLCofcarotenoids(lutein,beta-carotene),astaxanthinandtanninonsilicagelwithpetroleumether-diethylether-acetone15:�:2inatwin-troughchambersaturatedfor�0minatroomtemperature.Quantitativedeterminationbydensitometricabsorbancemeasurementat450nm.ThehRfvaluesofluteinintea,mulberry,andcassavaleafwere19,22,and19andcorrespondedtoluteinstandard.TheleastpolarzonehadanaveragehRfvalueof98,98,and96fortea,mulberry,andcassavaleaf,respec-tively,andwasidenticalwithbeta-carotenestandard.

foodanalysis,densitometry,quantitativeanalysis �0b

105052 A.ZEB*,M.MURKOVIC(*Institute forBiochemistry,GrazUniversityofTechnology,Graz,Austria; [email protected]): High-performance thin-layer chromatographic method for


monitoring the thermal degradation of ß-carotene in sunflower oil. J. Planar Chromatogr. 2�,�5-�9(2010).HPTLCofbeta-caroteneonsilicagel(prewashedwithmethanol)withpetroleumether -hexane - acetone2:�:1 in a saturated twin-troughchamber.Quantitativedeterminationbyabsorbancemeasurementat450nm.Linearitywasbetween100and600ng/band.LODandLOQwere0.11and0.�7ng/band,respectively.Averageintra-dayprecisionandinter-day-precisi-onwere0.54%and0.50%,respectively.

foodanalysis,HPTLC,quantitativeanalysis,densitometry �0b

10500� A.ZEBetal.,seesection1

32. Pharmaceutical and biomedical applications

10505� R.ARAVIND*,J.SAJAN,K.BINDU,A.BINDU(*Dept.ofPharmaceuticalScience,Cheruvan-doorCampus,Kottayam,Kerala,India):QuantitativedeterminationofquercetinpresentintheleavesofCinnamomummalabatrum(Burman)BusingHPTLCmethod.AbstractNo.259,61stIPC(2009).HPTLCofquercetininalcoholicextractsoftheleavesofCinnamomummalabatrumonsilicagelwithtoluene-acetone-formicacid�6:12:5.Quantitativeabsorbancemeasurementat254nm.Thealcoholicextractwasfoundtocontain10.02mg/gofquercetin.Thetotalflavo-noidcontentwasestimatedusingacolorimetricmethodwithaluminumchloride.ResultswereingoodcorrelationwiththeHPTLCmethod.

pharmaceuticalresearch,qualitycontrol,herbal,HPTLC,comparisonofmethods,quantitativeanalysis �2e

10508� AmeliaM.AVACHAT*,S.B.BHISE(*DepartmentofPharmaceutics,SinhgadCollegeofPhar-macy,OffSinhgadRoad,Vadgaon(Bk.),Pune-411041,India;[email protected]):Sta-bility-indicatingvalidatedHPTLCmethodforsimultaneousanalysisofrifabutinandisoniazidinpharmaceuticalformulations.J.PlanarChromatogr.2�,12�-128(2010).HPTLCofrifabutinandisoniazidonsilicagelwithdichloromethane-acetone-methanol20:7:2inatwin-troughchamberpreviouslysaturatedfor25min.Quantitativedeterminationbyabsorbancemeasurementat504nmforrifabutin(hRf84)andat262nmforisoniazid(hRf48).Thelinearityrangewas10-70and5-�5µg/mL,thecorrelationcoefficientwas0.9991and0.9989,theprecision(RSD,n=6)0.90%and0.71%,andprecisionondifferentdays(RSD,n=�)was0.89%and1.01%.LODwas180µgand90µg/zone,LOQwas540and270µg/zone,andthesystemsuitability(RSD,n=6)was1.41%and1.86%forrifabutinandisoniazid,respectively.

qualitycontrol,HPTLC,quantitativeanalysis,densitometry �2a

105054 D.BAHETI*,P.SHINDE,M.AGRAWAL,R.BANGAR(*SitabaiThiteCollegeofPharmacy,Pune, Maharashtra, India): Quantitative estimation of withanolide A in marketed polyherbalspansules.AbstractNo.C-�72,61stIPC(2009).HPTLCofwithanolideAinextractsandspan-sulesdosageformonsilicagelwith toluene-ethylacetate-formicacid8:6:1.Densitometricevaluationat254nm.WithanolideAwaswellseparatedwithanhRfvalueof14.Thelinearityrangewas40-200ng/band.

pharmaceuticalresearch,qualitycontrol,herbal,densitometry,HPTLC,quantitativeanalysis �2e

105055 G.BANSAL*,ReechaMADAAN,S.KUMAR(*S.D.CollegeofPharmacy,Barnala,Punjab,India):PharmacognosticinvestigationonActaeaspicataL.AbstractNo.C-274,61stIPC(2009).TLCofpetroleumetherandchloroformextractsofActaeaspicatarootsonsilicagelwithn-he-xane-chloroform9:1(forpetroleumetherextracts)andtoluene-ethylacetate-glacialaceticacid80:50:15:1(forchloroformextracts).Detectionbysprayingwith50%methanolicsulfuric


acidfollowedbyheatingat105°C.Petroleumetherextractsshowed2bands,whereaschloro-formextractsshowed�bands.

qualitycontrol,herbal,qualitativeidentification �2e

105056 BharatiBARMECHA*,D.GHANAWAT,P.SHINDE,SmitaSHELKE(*SitabaiThiteCollegeofPharmacy,Pune,Maharashtra,India):Simultaneousdeterminationofgallicacidandpiperinebyhigh-performancethin-layerchromatography.AbstractNo.C-��8,61IPC(2009).HPTLCofgallicacidandpiperineincombinedformulationonsilicagelwithtoluene-ethylacetate-for-micacid16:8:1.Quantitativeabsorbancemeasurementat�20nm.Themethodwaslinearintherangeof200-800ng/bandforgallicacidand50-�50ng/bandforpiperine.


105057 P.BHARATI,A.VINODINI.A.S.REDDY,P.S.DEVI*(DepartmentofPharmaceuticalSci-ences,AndhraUniversity,Visakhapatnam-5�000�,India;[email protected]):Developmentandvalidationofaplanarchromatographicmethodwithreflectancescanningdensitometryforquan-titativeanalysisofanastrozole in thebulkmaterialand in tablet formulations. J.PlanarChro-matogr. 2�, 79-8� (2010). HPTLC of anastrozole (2-[�-(1-cyano-1-methyl-ethyl)-5-(1H-1,2,4-triazole-1-yl-methyl)-phenyl]-2-methyl-propanenitrile)onsilicagel(prewashedwithmethanol)withtoluene-acetone-ammonia60:40:�inatwin-troughchamberpreviouslysaturatedfor20min.Quantitativedeterminationbyabsorbancemeasurementat200nm.Recoveryofanastrozolefrom1mg tablet formulations ranged from98.9 to101.5%. Intra-dayand inter-dayprecisionwere1.�4%and1.59%,respectively.LODandLOQwere71and214ng/band.Thecorrelationcoefficientfortheanastrozolecalibrationwas0.998�overtherangeof500-1500ng/band(peakarea).

qualitycontrol,densitometry,quantitativeanalysis,HPTLC �2a

105058 K.BOBER(DepartmentofAnalyticalChemistry,FacultyofPharmacy,MedicalUniversityofSilesia,4JagiellonskaStreet,PL-41-200,Sosnowiec,Poland,bober@sum):Densitometricana-lysisofselectedfluorquinolines.J.Liq.Chromatogr.Relat.Technol.��,778-785(2010).TLCofofloxacinandpefloxacinonsilicagelwithacetonitrile-formicacid-water40:�:7inchambersaturatedfor�0min.Quantitativedeterminationbydensitometryat295nmforofloxacinandat280nmforpefloxacin.TheregressionequationswereachievedusingcomputerprogramSTATI-STICA7.1.

qualitycontrol,densitometry,quantitativeanalysis �2a

105059 M.BOMBAYWALA*,D.MOHALE,A.CHANDEWAR(*P.WADHWANICollegeofPharma-cy,Yavatmal,Mah,India):BioassayguidedfractionationofLagenariasicerariaforantihyperlipi-demicactivity.InternationalSeminaronHerbalDrugResearch,PN-009(2009).BioassayguidedfractionationofLagenariasicerariawascarriedoutonasilicagelcolumnwithsolventsinascen-dingorderofpolarity.EachfractionobtainedwassubjectedtopreparativeTLConsilicagelwithn-butanol-methanol-water�:1:1.FourbandswithdifferentRfvalueswerecollectedandactivecompoundswereextractedandscreenedforantihyperlipidemicactivity.

pharmaceuticalresearch,traditionalmedicine,clinicalchemistryresearch,herbal,preparativeTLC �2e

105060 A.BORKAR*,S.MULGUND,A.GAJBHAR,K.JAIN(*SinhgadCollegeofPharmacy,Pune,Maharashtra,India):HPLC-PADandHPTLCmethodsforquantitativeandchromatographicfin-


gerprintanalysisofEmbellaribes(Vidanga)Churnaformulation.AbstractNo.F-10,61stIPC(2009).HPTLCofEmbellaribesChurnaformulationonsilicagelwithchloroform-ethylacetate-formicacid5:4:1inatwintroughchamber.Densitometricmeasurementofembelinat291nm.Themethodwaslinearintherangeof600-1800ng/bandwithrecoveryvalueof99.1-101.2%.TheformulationwasalsoanalyzedbyHPLCandresultswerefoundtobecomparable. p h a r -maceuticalresearch,qualitycontrol

herbal,densitometry,HPTLC,comparisonofmethods,quantitativeanalysis �2e

105061 P.CHANDRA*,A.RATHORE,L.SATHIYANARAYANAN,K.MAHADIK(*BharatiVidya-peethUniversity,PoonaCollegeofPharmacy,Pune,Maharashtra,India):Developmentofvalida-tedHPLCandHPTLCmethodsforsimultaneousdeterminationoflevocetirizinedihydrochlorideandmontelukastsodiuminbulkdrugandpharmaceuticaldosageform.AbstractNo.F-18,61stIPC(2009).HPTLCoflevocetirizinedihydrochlorideandmontelukastsodiuminbulkandtabletdosageformulationonsilicagelwithtoluene-ethylacetate-methanol-25%ammonia5:14:5:2.BothdrugswerewellresolvedwithhRfvaluesof�1forlevocetirizineand44formontelukast.Quantitativeevaluationat2�1nm.Themethodwaslinearintherangeof500-2500ng/bandforlevocetirizineand1000-5000ng/bandformontelukast.BothdrugswerealsoanalysedbyHPLConRP18columnandresultswerecomparablewithHPTLC.

pharmaceuticalresearch,qualitycontrol,HPTLC,densitometry,comparisonofmethods �2a

105062 K.DATTA,A.SINGH*,A.MUKHERJEE,B.BHAT,B.RAMESH,A.BURMAN(*Molecu-larOncologyLab,DaburResearchFoundation,Sahibabad,India,[email protected]):Ecliptaalbaextractwithpotentialforhairgrowthpromotingactivity.J.Ethnopharmacol.124,450-456(2010). HPTLC fingerprinting of Eclipta alba on silica gel with chloroform - ethanol - water�5:10:2.Densitometricevaluationat254nm.Majorcompoundsidentifiedwerecoumestantsandwedelolactone,withhairgrowthpromotingactivity.

traditionalmedicine,herbal,HPTLC,qualitativeidentification �2e

10506� R.DEEPA*,K.MADHURI,R.SHANMUGAM,G.SADAGOBAN(*J.S.S.CollegeofPhar-macy,Ooty,T.N.,India):EstimationofharmalineinPeganumharmalabyHPTLC.InternationalSeminaronHerbalDrugResearch,PN-011(2009).HPTLCofharmalineinalcoholicextractsofPeganumharmalaonsilicagelwithchloroform-acetone-diethylamine5:4:1inasatura-tedchamberat25°C.ThehRfofharmalinewas50.Quantitativedeterminationbyabsorbancemeasurementat�51nm.

pharmaceuticalresearch,herbal,densitometry,quantitativeanalysis �2e,17a

105064 A.DHOBI*,N.VEKARIYA,G.PATEL,R.DHOLAKIYA,C.SHASHTRY(*ShreeDhanvan-taryPharmacyCollege,Kim,Gujarat,India):Developmentandvalidationofanalyticalmethodfor simultaneous determination of telmisartan and amlodipine besylate in bulk and tablets byHPTLC.AbstractNo.F-15961st IPC(2009).HPTLCof telmisartanandamlodipinebesylateonsilicagelwithtetrahydrofuran-dichloroethane-methanol-25%ammonia�0:10:5:2.BothcompoundswerewellresolvedwithhRfvaluesof22and45fortelmisartanandamlodipinebe-sylaterespectively.Densitometricevaluationat�26nm.Themethodwasfoundtobelinearintherangeof1200-7200ng/bandfortelmisartanand400-1400ng/bandforamlodipinebesylate.

qualitycontrol,densitometry,HPTLC,quantitativeanalysis �2a

105065 TatjanaDJAKOVIC-SEKULIC*,V.DESPOTOVIC,G.USCUMLIC(*DepartmentofChemistry,Biochemistry,andEnvironmentalProtection,UniversityofNoviSad,FacultyofSciences,Trg


DositejaObradovica�,21000NoviSad,RepublicofSerbia; [email protected]):Quantitativestructure-retentionrelationshipsstudyof theretentiondataof5,5-disubsti-tutedhydantoins.J.PlanarChromatogr.2�,201-207(2010).TLCandHPTLCof175,5-disubs-tituted hydantoins on silica gel with ethyl acetate - toluene (with �0-60 % ethyl acetate) andacetonitrile-toluene(with�0-50%acetonitrile)andonRP18withmethanol-water(with56-80%methanol)andacetonitrile -water (with�0-60%acetonitrile)at roomtemperaturewithoutchambersaturation.DetectionunderUV254nm.Theeffectofthestructuresofthederivativesontheirretentioninbothnormalandreversed-phasemodeswasinvestigatedbyuseofQSRRandmoleculardescriptors.Cross-validationindicatedthebestmodelsarereliableQSRRmodels.

qualitativeidentification,postchromatographicderivatization �2a

105066 N.DUBEY*,N.DUBEY,R.MEHTA,A.SALUJA(*SophisticatedInstrumentationCenterforAppliedResearchandTesting,VallabhVidyaNagar,Gujarat,IndiaandDeviAhilyaVishwavi-dyalaya,SchoolofPharmacy,Indore,MadhyaPradesh,India;[email protected]):SelectivedeterminationofaconitineinpolyherbaloilscontainingAconitumchasmanthumusinghigh-performance thin-layer chromatography. J.AOAC Int. 9�, 1617-1621 (2010). HPTLC ofaconitineonsilicagelwithethylacetate-ethanol�:1at22°Cinasaturatedtwin-troughchamber.ThehRfvalueofaconitinewas��.Quantitativedeterminationbyabsorptionmeasurementat2�8nm.LODandLOQwas20and70ng/band,respectively.Thelinearitywithrespecttopeakareawasintherangeof�00to1800ng/bandwithanrof0.9991.Therepeatability(RSD)was0.85%;andtheinter-dayandintra-dayprecision(RSD)was1.01-1.�8and1.04-1.�4%,respectively.

qualitycontrol,herbal,traditionalmedicine,HPTLC,densitometry,quantitativeanalysis �2e

105067 S.DWIVEDI*,J.BORKAR,A.SAOJI,P.YEOLE(*InstituteofPharmaceuticalEducationandResearch,Wardha,Maharashtra,India):PhytochemicalscreeningandevaluationbyTLCandUVspectrophotometerofpolyherbaltablets.AbstractNo.C-495,61stIPC(2009).Screeningofdif-ferent phytoconstituents in a polyherbal tablet formulation.TLCofn-hexane, chloroformandmethanolextractsofthetabletsonsilicagelwithn-hexane-ethylacetate7:�;chloroform-me-thanol9:1,andchloroform-glacialaceticacid-methanol-water8:40:15:10.EvaluationunderUV254nmaswellasunderUV�66nmaftersprayingwithdifferentreagents:20%sulfuricacid,aniline-hydrogenphthalatereagent,anisaldehyde-sulfuricacidreagent,andvanillin-sulfuricacidreagentfor thedetectionofpiperineandandrographide, theactiveconstituentspresent informulations likeTefrolivForte tablets.Otherconstituents(tanninsetc.)wereanalyzedbyUVspectrophotometry.

pharmaceuticalresearch,qualitycontrol,herbal,densitometry,qualitativeidentification �2e

105068 M.FAIYAZUDDIN*,J.ALI,S.AHMAD,N.AHMAD,J.AKHTAR,S.BABOOTA(*Formula-tionResearchLaboratory,DepartmentofPharmaceutics,FacultyofPharmacy,JamiaHamdard,NewDelhi-110062,India,andDepartmentofPharmaceutics,FacultyofPharmacy,IntegralUni-versity,Lucknow-226026,UttarPradesh,India;[email protected]):Chromato-graphicanalysisoftrans-andcis-citralinlemongrassoilandinatopicalphytonanocosmeceuti-calformulation,andvalidationofthemethod.J.PlanarChromatogr.2�,2��-2�6(2010).HPTLCoftrans-citralandcis-citralinlemongrassoilonsilicagelwithtoluene-ethylacetate17:�inatwin-troughchambersaturatedfor15min(at25°Cand55%RH).Detectionbysprayingwithvanillin-sulfuricacidreagent.Quantitativedeterminationbyabsorbancemeasurementat595nm.Intra-dayandinter-dayprecisionwereevaluatedbyreplicate(n=6)analysisofsamples(trans-citralat450,900,and1800ng/band,andcis-citralat470,940,and1880ng/band).Thelinearrangewas225-�600ng/bandfortrans-citral,and470-�760ng/bandforcis-citral.Thecorrelationcoefficientrwas0.99��fortrans-citraland0.99�7forcis-citral.Intra-dayprecision(n=6)was<�.56and5.66%fortrans-andcis-citral,respectively.Inter-dayprecisionwasassessedtobe<�.47and5.52%fortrans-andcis-citralbyrepeatingtheintra-dayassayonthreedifferentdays.


Repeatabilityofsampleapplicationandpeak-areameasurementwas0.98%,determinedbyper-forming six replicate analysesof the sameband (1800ng/band trans-citral and1880ng/bandcis-citral).TheRSDofrecoveryoftransandcis-citralwasintheranges1.�6-�.25and1.64-�.47,respectively.

herbal,qualitycontrol,cosmetics,HPTLC,quantitativeanalysis,densitometry �2e

105069 M. GANDHIMATHI*, M. SARAVANA KUMAR, R. BAGHLA (*College of PharmacySRIPMS,Coimbatore,TN,India):RP-HPTLCmethodfortheinvitroestimationofedaravoneinhumanplasma.IPAConvention,2010,RA-PO0�.HPTLCofedaravoneinsamplesofhumanplasma(purifiedbyliquid-liquidextraction)onRP-18withn-butanol-methanol-diethylether1:8:1.ThecompoundwaswellresolvedwithanhRfvalueof81.Densitometricevaluationat240nm.Thelimitofdetectionandquantificationwas25ngand150ngrespectively.Thelinearitywas600-2400ng/band.Recovery(%)basedonanalysisofspikedsamplewasmorethan65%.pharmaceuticalresearch,clinicalchemistryresearch,HPTLC,densitometry,

quantitativeanalysis �2c

105070 A.GANTAIT,S.PANDIT,N.K.NEMA,P.K.MUKJERJEE*(*JadavpurUniversity,SchoolofNaturalProductStudies,Kolkata-7000�2,India;[email protected]):Quantificationofglycyrrhizin inGlycyrrhizaglabra extract byvalidatedHPTLCdensitometry. J.AOAC Int.9�,492-495 (2010).HPTLCofglycyrrhizinon silicagelwithchloroform-methanol -water1�0:72:15inatwin-troughchambersaturatedfor�0min.DetectionunderUV254nmandaftersprayingwithanisaldehyde-sulfuricacidreagent.ThehRfvalueofglycyrrhizinwas22.Quanti-tativedeterminationbydensitometryat254nm.Thelinearitywasbetween0.96-4.80µg/spot,thecorrelationcoefficientwasr=0.99904andthestandarddeviationwas2.52%.Averagerecoverywas99.6%.LOQandLODwas246and81ng/spot.

herbal,traditionalmedicine,HPTLC,densitometry,quantitativeanalysis �2e

105071 TatanaGONDOVÁ*,D.HALAMOVÁ,K.SPACAYOVÁ(*DepartmentofAnalyticalChemistry,Faculty of Science, P. J. Safárik University, Moyzesova 11, Kosice SK-040 ß1, Slovak Repu-blic; [email protected]):Simultaneousanalysisofnewantidepressantsbydensitometricthin-layerchromatography. J.Liq.Chromatogr.Relat.Technol.�1,2429-2441 (2008).TLCofcitalopram,sertraline,fluoxetine,andfluvoxamineonsilicagelwithacetone-benzene-25%ammonia10:9:1ina twin-troughchambersaturatedfor15min.DetectionunderUV254nm.ThehRfvaluewas28forfluoxetine,44forcitalopram,56forfluvoxamine,and68forsertraline(withastandarddeviationless than0.02%inallcases).Quantitativedeterminationbyabsor-bancemeasurementat240nm.Thecalibrationcurvewaslinearintherangeof500-5000ng/spotforallanalyzedcompounds;correlationcoefficientswere found tobemore than0.998foralldrugs(exceptthe0.991forfluvoxamine).LODwas40ng/spotforcitalopramand50ng/spotforfluoxetine,fluvoxamine,andsertraline,respectively.LOQwasfoundtobe1�0and160ng/spotforcitalopramandfluoxetine,respectively.Intra-assayprecision(%RSD)waswithintherangeof0.52-0.87%and1.�4-1.84%forcitalopramandfluoxetine,respectively,inter-dayprecisionfortheanalysesconductedonthreeconsecutivedayswasbelow1.8%and2.5%forcitalopramandfluoxetine,respectively.Therecoveries(RSD)ofcitalopramadfluoxetinewerefoundtobeintherangeof99.�-100.�%(0.8%)and99.4-100.4%(1.9%),respectively.


105072 AnnaGUMIENICZEK*,A,BERECKA(*MedicalUniversityofLublin, Jaczewskiego4,20-090Lublin,Poland;[email protected]):Quantitativeanalysisofglicazideandglipi-zideintabletsbyanewvalidatedandstability-indicatingRPTLCmethod.J.PlanarChromatogr.2�,129-1��(2010).TLCofglicazideandglipizideonRP18silicagelwith60%acetonitrilein


pH2.�phosphatebufferinanunsaturatedhorizontalchambersatroomtemperature.Detectionandquantitativedeterminationbyabsorbancemeasurementat215nm.Linearitywasintherangeof0.8-1.8µg/zoneforbothdrugsandthecorrelationcoefficientsrwere0.998forgliazide(hRf�8)and0.99�forglipizide(hRf51).LODandLOQwere50and200ng/zone,respectively,forglicazideand60and�00ng/zoneforglipizide.


10507� K.R.GUPTA*,M.R.TAJNE,S.G.WADODKAR(*BhoyarCollegeofPharmacy,NearDragonPalaceTemple,NewKamptee-441002,Dist:Nagpur(M.S.), India;[email protected]):Avalidatedhigh-performance thin-layer chromatographicmethod for thequantificationofsertralineintablets.J.PlanarChromatogr.2�,1�4-1�6(2010).TLCofsertralineonsilicagel(prewashedwithmethanol)with chloroform - ethyl acetate - triethylamine25:15:1 in a twin-troughchamberpreviouslysaturatedfor10minatroomtemperature.Quantitativedeterminationbyabsorbancemeasurementat279nm.ThehRfforsertralinewas40.Intra-dayprecisionandinter-dayprecisionwas99.84%RSDand99.92%RSD,respectively.Agoodlinearrelationshipbetweenresponse(peakarea)andamountwasobtainedovertherange2.7-7.9µg/band.

qualitycontrol,HPTLC,densitometry,quantitativeanalysis �2a

105074 D. HALAMOVA*, M. BADANICOVA,V. ZELENAK,T. GONDOVA, U.VAINIO (*Depart-mentofInorganicChemistry,FacultyofScience,P.J.SafarikUniversity,Moyzesova11,SlovacRepublic,[email protected]):Naproxendrugdeliveryusingperiodicmesoporous silicaSBA-15.Appl.Surf.Sci.256,6489-6494(2010).TLCofnaproxenreleasedfromaSBA-15me-soporoussilicadrugdeliverysystemonsilicagelwithbenzene-tetrachloromethane-aceticacid7:1:1.Quantitativedeterminationbyabsorbancemeasurementat260nmatdifferenttimeinter-vals.ThehRfvalueofnaproxenwas50.

pharmaceuticalresearch,densitometry,quantitativeanalysis �2a

105075 PurnimaHAMRAPURKAR*,S.PAWAR,M.PHALE(*DepartmentofPharmaceuticalAnaly-sis,Prin.K.M.KundnaniCollegeofPharmacy,JoteJoyBuilding,RambhauSalgaonkarMarg,CuffeParade,Colaba,Mumbai-400005,India;[email protected]):QuantitativeHPTLCanalysisofphyllanthininPhyllanthusamarus.J.PlanarChromatogr.2�,112-115(2010).HPT-LCofphyllanthinonsilicagel(prewashedwithmethanol)withhexane-toluene-ethylacetate2:2:1inatwin-troughchambersaturatedat25-�0°Cand40-50%relativehumidity.Quantitativedeterminationbyabsorbancemeasurementat206nm.LODwas70ng/mL,LOQ200ng/mL.Thelinearcalibrationrangewas200-1200ng/mL.Repeatability(RSD,n=�)was0.18-0.59%withacorrelationcoefficientof0.999.Intra-dayandinter-dayprecisionstudiesshowedtheCVwaslessthan2.0%,indicatingthemethodwasprecise:%RSDat200,600,and1200ng/mLwasbetween0.4�and1.51%forintra-dayprecision,andbetween0.59and1.7�%forinter-dayprecision.Theintra-dayrecoveryfor200,600and1200ng/mLwasbetween102.�%and99.9%,andtheinter-dayrecoverybetween102.5%and99.9%,respectively.

traditionalmedicine,herbal,HPTLC,densitometry,quantitativeanalysis �2e

105076 R.JAYAPRAKASAM*,M.SASIKALA,M..GANDHIMATHI,M.SUKUMAR,T.RAVI(*Col-legeofPharmacy,SriRamkrishnaInst.ofParaMed.Science,Coimbatore,T.N.,India):HPLC&HPTLCfingerprintingandin-vitroantioxidantstudiesofvariousextracts,isolatedcompoundsandformulationsofEugeniajambolanaLam.InternationalSeminaronHerbalDrugResearch,PN-01� (2009).HPTLCof ethyl acetate extractsof seedsofEugenia jambolanaon silicagelwithchloroform-acetone-formicacid150:��:17.TheantioxidantactivityofdifferentextractwasassessedbyDPPHmethod.Theproposedmethodwasappliedtoplantextractaswellastopolyherbalformulation.Theethylacetateextractwasfoundtohavegoodantioxidantactivity.For


HPLCfingerprintprofilingaRP18columnwasusedwith80%methanolasmobilephase.

herbal,comparisonofmethods,qualitativeidentification �2e

105077 A. JOHNSON, A. KUMAR, S. RASHEED, S. CHANDRIKA, A. CHANDRASEKHAR, S.BABY*, A. SUBRAMONIAM (*Phytochemistry and Phytopharmacology Division, TropicalBotanicalGardenandResearchInstitute,Pacha-Palode,Kerala,India,[email protected]):An-tipyretic,analgesic,anti-inflammatoryandantioxidantactivitiesof twomajorchromenesfromMelicopelunu-ankenda.J.Ethnopharmacol.1�0,267-271(2010).HPTLCofevodioneandlep-tonolfromtheleavesandinfluorescencesofMelicopelunu-ankendaonsilicagelwithchloro-form-methanol1:1.Detectionbysprayingwithanisaldehyde-sulfuricacidreagent,followedbyheatingat105°Cfor5min.Quantitativedeterminationbyabsorbancemeasurementat580nm.

traditionalmedicine,herbal,HPTLC,quantitativeanalysis,densitometry �2e

105078 V.KADAM*,VarshaJADHAV,SapanaKAMBLE,A.PAHADE(*BharatiVidyapeeth‘sCollegeofPharmacy,NaviMumbai,Maharashtra,India):DevelopmentandvalidationofHPTLCmethodfordeterminationof�-hydroxyandrostane(16,17-C)(6-methyl2‘-1-hydroxy-isopropene-1-yl)-4,5,6H-pyraninherbalformulation.AbstractNo.C-2�1,61stIPC(2009).AnHPTLCmethodisreportedfordeterminationof�-OH-androstane-(16,17-C)(6-methyl-2-1-hydroxy-isopropene-1-yl)-4,5,6H-pyran,aphytoconstituentofEugeniajambolana.Thecompoundwasisolatedbyethanolicextraction,identifiedbymeltingpoint,IR,andNMR,andusedasmarker.HPTLConsilicagelwithtoluene-ethylacetate17:�.Densitometricevaluationat�66nm.Themethodwaslinearintherangeof1000-5000ng/band.ItcanbeusedforroutinequalitycontrolofEugeniajambolanaseedsandherbalformulation.

pharmaceuticalresearch,qualitycontrol,herbal,HPTLC,quantitativeanalysis �2e

105079 A.KHATIB*,A.C.HOEK,S.JINAP,M.Z.I.SARKER,I.JASWIR,R.VERPOORTE(*Centerof Excellence for Food Safety Research, Faculty of Food Science andTechnology, UniversityPutraMalaysia,4�400Serdang,SelangorDarulEhsan,Malysia;[email protected]):Ap-plicationoftwodimensionalthinlayerchromatographypatterncomparisonforfingerprintingtheactivecompoundsintheleavesofVitextrifoliaLinnpossessinganti-tracheospasmolyticactivity.J.Liq.Chromatogr.Relat.Technol.��,214-224(2010).TLCofethanolicextractsoftheleavesofVitextrifoliaonsilicagelwithchloroform-methanol9:1inthefirstdirectionandethylacetate-chloroform-methanol7:7:11intheseconddirection.DetectionunderUVlightat254and�66nmandbysprayingwithanisaldehyde-sulfuricacidreagent.

traditionalmedicine,herbal �2e

105080 V.KUMAR*,S.VARGHESE,H.JOHN(*Dept.ofPharmaceuticalAnalysis,CollegeofPharma-cy,SRIPMS,Coimbatore,T.N.,India):DevelopmentofvalidatedHPTLC&HPLCmethodsforestimationofciticolinesodiumintabletdosageform.IPAConvention,2010,RA-PO�5.HPTLCofciticolinesodiumintabletformulationonsilicagelwithchloroform-methanol-water�:7:�.ThecompoundwaswellresolvedwithanhRfvalueof5�.Densitometricmeasurementat280nm.Themethodwaslinearintherangeof�00-900ng/band.HPLCanalysiswasperformedonRP18columnusing1%formicacid-methanol19:1.Resultsobtainedwitheithermethodwerecomparable.

pharmaceuticalresearch,qualitycontrol,comparisonofmethods,quantitativeanalysis,densitometry �2a

105081 P. LAHORKAR*, K. RAMITHA,V. BANSAL, D.B.ANANTHA NARAYANA (*Gerbal Re-searchLaboratory,HindustanUnileverResearchCentre,64MainRaod,Whitefield,Bangalore


560066,India):Acomparativeevaluationofmedicatedoilspreparedusingayurvedicandmo-difiedprocesses.Ind.J.Pharma.Science7161,656-662(2009).MedicatedoilspreparedbothbyAyurvedicaswellasmodifiedprocesswereevaluatedforfingerprintprofilingbyHPLCandHPTLC.HPTLCofmethanolicextractsoftheoilsonsilicagelwithchloroformmethanol9:1andtoluene-ethylacetate4:1forgeneralfingerprintprofiling;withtoluene-ethylacetate-for-micacid5:4:1forflavonoidsandwithtoluene-ethylacetate-diethylamine7:2:1foralkaloids.Evaluationunder254nmand�65nm.DetectionbytreatmentwithNP-PEGreagentforflavono-idsandDragendorffreagentforalkaloids.

pharmaceuticalresearch,traditionalmedicine,herbal,postchromatographicderivatization,qualitativeidentification,HPTLC �2e

105082 B.LUKAS,C.SCHMIDERER,U.MITTEREGGER,J.NOVAK*(*InstituteofAppliedBotanyandPharmacognosy,DepartmentforFarmAnimalsandVeterinaryPublicHealth,UniversityofVeterinaryMedicine,Vienna,Austria,[email protected]):Arbutininmarjoramandoregano.FoodChem.121,185-190(2010).TLCofarbutinintheleavesandflowersofOriganummajorana andOriganumvulgareon silicagelwith ethyl acetate -methanol -water 77:1�:10.Detectionbysprayingwith2.5%dibromchinochlorimidesolutioninethanol.ThehRfofarbutinwas47.

foodanalysis,herbal,qualitativeidentification �2e

105084 ÁgnesM.MÓRICZ*,E.TYIHÁK,P.G.OTT(*PlantProtectionInstitute,HungarianAcademyof Sciences, Herman O. u. 15, 1022 Budapest, Hungary; [email protected]): Usefulness oftransgenicluminescentbacteriaindirectbioautographicinvestigationofchamomileextracts.J.PlanarChromatogr.2�,180-18�(2010).TLCofchamomileextractsonsilicagelwithbenzene-ethylacetate19:1orchloroform-methanol-water40:10:1inanunsaturatedchamber.Detectionat254and�66nm.ForbioautographicevaluationbioluminescentBacillussubtilisorPseudomo-nassyringaepv.maculicolawereused;visualizationbydyeareagentwasachievedbydippingtheplateinanaqueoussolutionofMTT.Quantitativedeterminationbydensitometricscanningat�00nm(beforebiologicaldetection)orat590nm(aftervisualizationofthebioautogramwithMTT).

qualitycontrol,herbal,traditionalmedicine,qualitativeidentification,densitometry,quantitativeanalysis,bioautography �2e

105085 A.MAKOWSKI*,E.ADAMEK,W.BARAN(*MedicalUniversityofSilesia,FacultyofPhar-macy,4JagiellonskaStreet,41-200Sosnowiec,Poland;[email protected]):UseofphotocatalyticreactionstovisualizedrugsinTLC.J.PlanarChromatogr.2�,84-86(2010).TLCof 18 drugs (6 antibiotics [benzyl-penicillin procaine, benzyl-penicillin potassium, penicillicacid,tetracyclinehydrochloride,oxytetracylinehydrochloride,chlortetracyclinehydrochloride],2 analgesics [aminophenazone, salicylamide], 2 anaesthetics [phenazone, procaine hydrochlo-ride],andoneeachofanti-rheumatic[penicillamine],anti-inflammatory[metamizolesodium],antitussive [codeinephosphate], broncholytic [aminophylline], spasmolytic [papaverinehydro-chloride], hypnotic [phenobarbital], sympathomimetic [ephedrine hydrochloride] and vitamin[ascorbicacid]drugs)onsilicagelwithbutanol-anhydrousaceticacid-water�:1:1.Detectionbysprayingwith10mLeachofasolutionof(A)0.25gtitanumdioxidein0.1mol/Lpotassiumpermanganate,(B)0.25gtitaniumdioxidein1.0mol/Lpotassiumiodide,(C)0.25gtitaniumdioxidein1.0mol/Lpotassiumbromide,whichwasthebest(mostsensitive)one,and(D)0.25gtitaniumdioxidein1.0mol/Lpotassiumchloride.AftersprayingwithreagentsCandD,plateswereilluminatedfor10min,sprayedwith0.1mol/Lsilvernitratesolutionandilluminatedagainfor�min.InallexperimentstheTLCplateswereilluminatedbyuseofUVlampswiththeradia-tionat�66nm.LODsformostofthedrugsstudiedwereintherange0.2-0.5µg/spot.

qualitativeidentification �2a


105086 AsthaMEHTA*,A.THAKER(*DepartmentofPharmaceuticalChemistry,SchoolofPharmacyandTechnology Management, NMIMS University,Vile Parle-W Mumbai-400056, India; [email protected]):ValidatedHPTLCmethodforassayofprednisoloneintabletsandcompa-risonwithpharmacopoeialmethods.J.PlanarChromatogr.2�,208-211(2010).HPTLCofpred-nisolone (andhydrocortisoneas impurity)onsilicagel (prewashedwithmethanol)withchlo-roform-methanol19:1inatwin-troughchamberpreviouslysaturatedfor�0min.Quantitativedeterminationbyscanningdensitometryat250nm.Linearitywasintherangeof2-10µg/band(r=0.9967,calculatedviapeakarea).LODandLOQwas200and600ng/spot, respectively.Recoverywas100.0%forprednisolone.Repeatabilitywas0.74%andtheinter-day(n=6)andintra-day(n=12)precisionwas2.6and2.7%,respectively.

qualitycontrol,comparisonofmethods,densitometry,HPTLC,quantitativeanalysis �2a

105087 B. MEHTA*, S. MORGE (*Dept. of Chem. University of Mumbai, Santacruz (E), Mumbai,400098, India):Simultaneousdeterminationof irbesartanandhydrochlorothiazidebyHPTLCmethod. Indian Drugs 47(2), 71-74 (2010). HPTLC of irbesartan and hydrochlorothiazide onsilicagelwithacetone-chloroform-ethylacetate-methanol6:6:6:1.Theplateswereprecondi-tionedfor10mininasaturatedchamberpriortodevelopment.ThehRfvalueofirbesartanwas27andofhydrochlorothiazide�7.Thelinearitywas1500-9000ng/bandand125-750ng/bandforirbesartanandhydrochlorothiaziderespectively.Theaveragerecoveryforbothdrugswas99.4-99.5%.

pharmaceuticalresearch,qualitycontrol,densitometry,quantitativeanalysis �2a,17c

105088 SigridMENNICKENT*,J.CONTRERAS,C.REYES,M.VEGA,M.DEDIEGO(*DepartmentofPharmacy,FacultyofPharmacy,UniversityofConcepción,P.O.Box2�7,Concepción,Chile;[email protected]):Validated instrumental planar chromatographic method for quantificationoffluphenazinehydrochlorideininjections.J.PlanarChromatogr.2�,75-78(2010).HPTLCoffluphenazinehydrochlorideonsilicagel(prewashedwithmethanol)withmethanol-water9:1inasaturatedtwin-troughchamber.Quantitativedeterminationbyabsorbancemeasurementat�06nm.Linearitywasintherangeof100to500ng/µLwithacorrelationcoefficientof0.998.LODandLOQwere1.45and4.40ng/zone,respectively.Intra-assayandinter-assayprecision,expressedas relative standarddeviation (RSD),were in the range0.7�-1.77%(n=�)and1.18-1.86%(n=9), respectively.Recoveryoffluphenazinehydrochloridewasbetween98.�and101.5%,withRSDnothigherthan1.87%.Themethodwasselectiveforfluphenazinehydrochlorideandthepreservativesintheinjections.


105089 E.MINCSOVICS*,N.TABANCA,Á.M.MÓRICZ,D.E.WEDGE,E.TYIHÁK(*OPLC-NITLtd,AndorStreet60,1119Budapest,Hungary,andCorvinusUniversity,FacultyofHorticulturalSciences, Dept. Genetics and Plant Breeding, Budapest, Hungary; [email protected]):PreliminaryinvestigationofOriganumonitesessentialoilbyoverpressuredlayerchroma-tographyandBioArena.J.PlanarChromatogr.2�,225-226(2010).OPLCoforeganooilcom-ponents (carvacrol, thymol, and linalool) on silicagelwithdichloromethane.DetectionunderUV254nm,bysprayingwithvanillin-sulfuricacidreagent(0.1gvanillin,100mLethanol,and2.2mL95-98%sulfuricacid)andheatingat110°Cfor�min,andintheBioArenasystem(thedrieddevelopedplatesweredippedfor10sintoanaqueouscellsuspensionofthesoilbacteriaBacillussubtilisandincubatedfor2hat100%rel.humidityand�0°C).Visualizionofanti-microbialcompoundswasperformedbyimmersingtheplatesfor5sinanaqueoussolutionofMTTreagent(80mgMTTand100mgTritonX-100in100mLwater).Thelayerswerefurtherincubatedandanddocumented.

herbal,foodanalysis,traditionalmedicine,qualitativeidentification �2e


105090 H.MISTRY*,S.SHUKLA,N.PRAJAPATI,B.JOGI(*InstituteofScienceandTechnologyforAdavancedStudiesandResearch,Gujarat,India):StandardizationofPanchkolChurnabyHPT-LCmethodforthedeterminationofpiperine,plumbagineandzingiberine.AbstractNo.C-258,61stIPC(2009).HPTLCofpiperine,plumbagineandzingiberineinPanchkolChurna,anayu-rvedicpreparationusedforanorexia,distensionandabdominalpain.HPTLConsilicagelwithtoluene-ethylacetate7:�.Densitometricevaluationat�40nmforpiperine,andat420nmforplumbagineandzingiberine.ThehRfvalueofpiperine,zingiberineandplumbaginewas�1,75,and84.


105091 P.NIRALI*,K.MANVITHA,K.SALMA,A.SHABARAYA(*SrinivasCollegeofPharmacy,Mangalore,India):DeterminationofandrographolideinAndrographispaniculataextractswithandwithouthumanserumbyHPTLC.AbstractNo.C-161,61stIPC(2009).AnHPTLCmethodisreportedforestimationofandrographolidesbitterprinciplesinAndrographispaniculata,po-pularlyknownaskalmegh.HPTLCofmethanolicandwaterextractsonsilicagelwithchloro-form-methanol7:1inasaturatedtwintroughchamber.Quantitativeevaluationbyabsorbancemeasurementat2�1nm.Themethodwasfoundtobelinearintherangeof1-5µg/band.Bothextractswerefoundtocontainandrographolides.Maximumyieldsofandrographolideswereob-servedinextractspreparedbyrefluxing.

pharmaceuticalresearch,qualitycontrol,HPTLC,densitometry,quantitativeanalysis �2e

105092 A. ÖZTUNC*,A. ÖNAL, S. E.TOKER (*Istanbul University, Faculty of Pharmacy, Depart-mentofAnalyticalChemistry,Istanbul,Turkey;[email protected]):Detectionofmetham-phetamine,methylenedioxymethamphetamine,and�,4-methylenedioxy-N-ethylamphetamineinspikedplasmabyHPLCandTLC.J.AOACInt.9�,556-561(2010).TLCofmethamphetamine(MA), �,4-methylenedioxymethamphetamine (MDMA), and �,4-methylenedioxy-N-ethylam-phetamine(MDEA)onsilicagelwithhexane-chloroform1:9andHPTLConcyanophasewithbenzene-diethylether-petroleumether(40-60°)-acetonitrile-ethylmethylketone4:7:7:1:1withchambersaturationfor�0min.ThehRfvalueofMA,MDMA,andMDAEonsilicagelwere28,2�,and�6,repectively,andoncyanophase�5,�0,and40,respectively.LODonsilicagelwere0.8,0.6,and1.2µg/mL(inplasma)forMA,MDMA,andMDEArespectively.

doping,toxicology,HPTLC �2a

10509� H.A.PANAHI*,A.RAHIMI,E.MONIRI,A. IZADI,M.M.PARVIN(*DepartmentofChe-mistry,CentralTehranBranch,IslamicAzadUniversity,Tehran,Iran;[email protected]):HPTLCseparationandquantitativeanalysisofaspirin, salicylicacid, andsulfosalicylicacid.J.PlanarChromatogr.2�,1�7-140(2010).HPTLCofaspirin,salicylicacid,andsulfosa-licylicacidonsilicagel(prewashedwithmethanol-chloroform1:1andimpregnatedwith2%boricacidinethanol)withchloroform-methanol-ammonia-water120:75:2:6inachamberpreviouslysaturatedat25°Cfor�0min.Detectionandquantitativedeterminationbydensito-metryat254nm.ThehRfofaspirin,salicylicacid,andsulfosalicylicacidwere81,61,and24,respectively.Thelinearrangewas100-1000ng/bandforallthreecompounds,andthecorrelationcoefficientsrwere0.97,0.94,and0.95,respectively.LOQwere12�,95,and61ng/band,respec-tively,andtherespectiveLODwere�7,�7,and18ng/band.


105095 N.PATEL*,D.MODI,B.SHAH,P.RACHH(*VidyabharatiTrustCollegeofPharmacy,Surat,Gujarat,India):EstimationofellagicacidinEugeniajambolanaLamseedalcoholicextractbyHPTLCmethod.AbstractNo.C-497,61stIPC(2009).HPTLCofellagicacidinseedsofEu-


geniajambolanaLamonsilicagelwithethylacetate-glacialaceticacid-formicacid-water100:11:11:27.Densitometricevaluationat254nm.Themethodwaslinearintherangeof200-1200ng/band.Thealcoholicseedextractcontained11.0�%ofellagicacidand21%of totaltannin(measuredbychemicalmethod).


105094 PrachiPATEL(M.P.PatelCollegeofPharmacy,Kheda,Gujarat,India):Quantificationofvasi-cineandpiperineinpolyherbalformulation.AbstractNo.C-47,61stIPC(2009).HPTLCofpi-perineandvasicineinpolyherbalcoughformulationsonsilicagelwithdioxane-toluene-ethylacetate -methanol - 25%ammonia15:20:10:10:�.Quantitativedeterminationby absorbancemeasurementat�04nm.Themethodwasvalidatedforaccuracy,precision,LOD,LOQ,linearityandspecificityandwasfoundtobelinearintherangeof2-10µg/bandforbothvasicineandpi-perine.

pharmaceuticalresearch,qualitycontrol,herbal,HPTLC,densitometry �2e

105096 S.PATHAN*,S.ALAM,G.JAIN,S.ZAIDI,S.AKHTER,D.VOHORA,R.KHAR,F.AHMAD(*Department of Pharmaceutics, Faculty of Pharmacy, Hamdard University, New Delhi,India,[email protected]):Quantitativeanalysisofsafranalinsaffronextractandnano-particleformulationbyavalidatedhigh-performancethin-layerchromatographicmethod.Phy-tochem.Anal.21,219-22�(2010).HPTLCofsafranalinsaffronextractandinasafranal-loadednanoparticleformulationonsilicagelwithn-hexane-ethylacetate9:1.Quantitativedetermina-tionbyabsorbancemeasurementat�10nm.ThehRfofsafranalwas51.Linearitywasbetween0.5and5.0µg/zone.Theintra-dayandinter-dayprecisionswere1.08-2.17and1.86-�.47%,re-spectively.LODwas50ng/zonewhileLOQwas150ng/zone.Theaveragerecoverywas99.9%.Theproposedmethodprovidessignificantadvantages in termsofgreaterspecificityandrapidanalysis.

pharmaceuticalresearch,herbal,HPTLC,densitometry,quantitativeanalysis �2a

105097 R.PIETRAS*,D.KOWALCZUK(*DepartmentofMedicinalChemistry,MedicalUniversityofLublin,4JaczewskiegoStr.,20-090Lublin,Poland;[email protected]):RP-TLCseparationof antiarrhythmicdrugs.Densitometric analysis offlecainide in tablets. J.PlanarChromatogr.2�,65-69(2010).TLCofsomeantiarrhythmiccompounds(disopyramidephosphate,verapamilhydrochloride,flecainideacetate,mexiletinehydrochloride,andtocainidehydrochloride)onRP8andRP18silicagelwithorganic-aqueousmobilephasescontainingcitrateoracetatebuffersatdifferentpHinahorizontalchamber.Thebestseparationofindividualandmixeddrugstandardswasachievedwithtetrahydrofuran-citratebuffer(pH4.45)�:7.Flecainideacetatewasidenti-fiedandquantifiedbyUVdensitometryat225and�10nm.Linearrelationshipswereobtainedbetweenpeakheightorpeakareaandamountintherange6.0to12.0µg/spot,thecorrelationco-efficientrwas0.990.Precision(RSD1.1-5.9%)andaccuracy(96.2-10�.6%)weresatisfactory.

pharmaceuticalresearch,qualitycontrol,densitometry,quantitativeanalysis �2a

105098 S.PRASAD*,S.HEMALATHA,T.THITE,M.KRISHNAN(*Dept.ofPharmaceutics,InstituteofTechnology,BanarasHinduUniv.,Varanasi,U.P.,India):Identificationandquantificationofwithaferin-A indifferent fractionsofWithania coagulansdunal byTLCandHPTLCmethod.AbstractNo.C-97,61st IPC (2009).Chromatographicmethodsare reported for identification(TLC) and quantification (HPTLC) of withaferin-A in methanolic and chloroform extract ofdriedfruitsofWithaniacoagulans.Chromatographicseparationonsilicagelwithtoluene-ethylacetate - formic acid5:5:1.The identificationofwithaferin-A inboth chloroformandmetha-nolicextractswasperformedbycomparisonofhRf valuesandUVabsorbancemaxima (209


nm). Quantification was performed by absorbance measurement at 540 nm after spraying thedevelopedplatewithLiebermann-Burchardreagent.Methanolicextractsandchloroformextractscontained�.67mg/gand2.10mg/gofwithaferin-A,respectively.Nowithaferin-Awasfoundinhydroalcoholicextracts.

pharmaceuticalresearch,qualitycontrol,densitometry,HPTLC,quantitativeanalysis �2e

105099 AlinaPYKA*,D.RUSEK,P.BOCHENSKA,D.GURAK(*DepartmentofAnalyticalChemistry,FacultyofPharmacy,MedicalUniversityofSilesia,4JagiellonskaStreet,PL-41-200Sosnowiec,Poland;[email protected]):UseofRP-TLCandtheoreticalcomputationalmethodstocomparethelipophilicityofsalicylicacidanditsderivatives.J.Liq.Chromatogr.Relat.Technol.��,179-190(2010).TLCofsalicylicacidanditsderivatives,namelyacetylsalicylicacid,salicylanilide,salicylaldehyde,salicylamide,salicylhydroxamicacid,methylsalicylate,phenylsalicylate,�,5-dinitrosalicylic acid, 2,5-dihydroxysalicylic acid, �-aminosalicylic acid, 4-aminosalicylic acid,and5-aminosalicylicacid,onRP8,RP18andHPTLConRP18andcyanophasewithmethanol-water; thecontentofmethanol inmobilephasewasgraduallyvariedby5%from20-100%.Developmentinachambersaturatedfor15min.Quantitativedeterminationbyscanningdensito-metryinabsorptionmodeattherespectiveabsorptionmaximum.ThehRfvalueswererecalcula-tedontheRMvalues.ThechromatogramswererepeatedintriplicateandmeanhRfvalueswerecalculated.TheresultsindicatethatthechromatographicparameteroflipophilicitydeterminedonRP8andcyanophasemaybeusedasameasureoflipophilicityoftheinvestigatedsalicylicacidanditsderivatives.

HPTLC,quantitativeanalysis,densitometry �2a

105100 AlinaPYKA*,P.BOCHENSKA(*DepartmentofAnalyticalChemistry,FacultyofPharmacy,MedicalUniversityofSilesia,4JagiellonskaStreet,PL-41-200Sosnowiec,Poland;[email protected]):ComparisonofNP-TLCandRP-TLCwithdensitometrytoquantitativeanalysisofibu-profeninpharmaceuticalpreparations.J.Liq.Chromatogr.Relat.Technol.��,825-8�6(2010).NP-TLCof ibuprofen standard and in tablet extractson silicagel (prewashedwithmethanol)withn-hexane-ethylacetate-aceticacid150:50:7andRP-TLConRP18withmethanol-water9:1inasaturatedtwin-troughchamber.Quantitaitvedeterminationbyabsorbancemeasurementat200and224nmforNP-TLCandRP-TLCanalysis,respectively.ThehRfvalueswere61and67.Linearitywasbetween2.50-12.50and2.50-12.50µg/spot,thecorrelationcoefficient(r)was0.998and0.994.LODwas0.60and1.00µg/spot,andLOQ12.50and12.50µg/spot.Recoverywas98.2%and96.1%,withastandarddeviationof1.21and1.45,andRSD1.28and1.81forNP-TLCandRP-TLC,respectively.


105101 BHARGAVI,R.SHANMUGAN*,K.MADHURI(*VelscollegeofPharmacy,Dept.ofPhar-maceutics, Chennai, India): Estimation of withanolide-A in nicandra physaloides by HPTLC.InternationalSeminaronHerbalDrugResearch,PN-008(2009).HPTLCofwithaferin-AfromNicandraphysaloidesonsilicagelwithtoluene-ethylacetate-formicacid10:�:1.ThehRfva-luewas14.Quantitativedeterminationbyabsorbancemeasurementat21�nm.

pharmaceuticalresearch,densitometry,quantitativeanalysis �2e

105102 Tirumala RAJESH*, K.S. LAKSHMI, S. SHARMA, P. D. REDDY, S. LAKSHMI (*Depart-mentofPharmaceuticalAnalysis,SRMCollegeofPharmacy,SRMUniversity,Kattankulathur-60�20�,TamilNadu, India, [email protected]):Useof avalidated stability-indica-tingHPTLCmethodtostudythedegradationofrimonabant.J.PlanarChromatogr.2�,148-155(2010).HPTLCofrimonabantanddegradationproductsonsilicagel inahorizontalchamberwithmethanol-water7:�.AcompactbandwasobtainedathRf71.Quantitativedetermination


byabsorbancemeasurementat250nm.Linearregressionanalysisofcalibrationdatarevealedgoodlinearrelationshipwithr=0.9985inthelinearworkingrangeof100-800ng/band.


10510� T.RAJKUMAR*,B.SINHA(*Dept.ofPharmaceuticalSciences,BirlaInstituteofTechnology,Mesra,Jharkhand,India):ChromatographicfingerprintanalysisofbudmunchiaminesinAlbiziabyHPTLCtechnique.AbstractNo.C-45�,61stIPC(2009).Fingerprintanalysisofbudmunchia-mines,themainconstituentsinAlbiziaamara.Driedpowderedleaveswereextractedwithpetro-leumether(60-80°C),chloroform,ethylacetateand90%methanolbymacerationfor48h.TLConsilicagelwithchloroform-methanol19:1.Zones2,4and8correspondedtothemarcocylicalkaloidsbudmunchiamineA,B,andC,whichwasconfirmedbyFTIR,NMRandMS.

pharmaceuticalresearch,qualitycontrol,herbal,densitometry �2e

105104 J. RAO*, K. CHAUHAN, K. MAHADIK (*Poona College of Pharmacy, BharatiVidyapeethUniversity, Pune, Maharashtra, India):Validated high-performance thin-layer chromatographicmethod for determination of diacerein in the presence of degradation products formed underICH-recommendedstressconditions.AbstractNo.F-2�461stIPC(2009).Astability-indicatingHPTLCmethod is reported forestimationofdiacerein in thepresenceof itsdegradationpro-ducts.HPTLConsilicagelwith toluene-ethylacetate - formicacid50:�0:1.ThehRf valueofdiacereinwas�9.Densitometricanalysisat254nm.Themethodwasfound tobe linear inthe rangeof100-600ng/band.Themethodwasvalidated regarding stability.The samplewassubjectedtodifferentstressconditions(acid/basehydrolysis,oxidation,photolysis,thermal)andshowedextensivedegradationinalkalinemediumandmilddegradationunderacidicandoxida-tiveconditions.Themethodallowedseparationofdiacereinfromdifferentdegradationproducts.

pharmaceuticalresearch,qualitycontrol,densitometry,HPTLC,quantitativeanalysis �2a

105105 SuparnaROY*,B.SINHA,MANIK(*BirlaInstituteofTechnology,Ranchi,Jharkhand,India):EstimationofconstituentsfrommethanolicextractofAloeverabyHPTLCtechnique.AbstractNo.C-�7,61stIPC(2009).HPTLCofmethanolicleafextractsofAloevera(afterpurificationwithpetroleumether(60-80°C))onsilicagelwithtoluene-ethylacetate-glacialaceticacid-methanol4:18:1:4.UnderUV254nmsixbandswithhRfvaluesof12,26,�4,44,62,and84wereobserved.ThesebandscorrespondtowellknownconstituentsofAloevera:aloeresinhRf25,barbaloinhRf��,aloeemodinhRf4�,emodinhRf6�.ThebandswithhRfvaluesof12and84couldnotbeidentified.Thereportedfingerprintprofilingcanserveaspotentialtechniqueforauthentificationandbatchtobatchconsistencyofherbaldrugs.

pharmaceuticalresearch,qualitycontrol,herbal,HPTLC,densitometry,qualitativeidentification,quantitativeanalysis �2e

105106 P.K.SAINI,R.M.SINGH*,S.C.MATHUR,G.N.SINGH,C.L.JAIN,R.K.KHAR,A.HAFEEZ(*IndianPharmacopoeiaCommission,MinistryofH.andF.W.,GovernmentofIndia,Rajnagar,Ghaziabad(U.P.)-201002,India;[email protected]):AsimpleandsensitiveHPTLCmethodforquantitativeanalysisofartemetherandlumefantrineintablets.J.PlanarChromatogr.2�,119-122(2010).HPTLCofartemetherandlumefantrineonsilicagelwithn-hexane-ethylacetate4:1atroomtemperatureinatwin-troughchambersaturatedfor15min.Quantitativede-terminationbydensitometricscanninginreflectancemodeat�57nm.Themethodislinear(r2>0.995),precise(RSD<2%),accurate(averagerecoveryof100.5%forartemetherand99.5%forlumefantrine),specific,androbust.LODandLOQforartemetherwere50and150ng/band,respectively,andthoseforlumefantrinewere�00and900ng/band,respectively.


CAMAGBIBLIOGRAPHYSERVICE No.105�0

105107 L. SAWANT*, N. PANDITA (*School of Pharmacy and Technology Management, NMIMSUniversity,Mumbai,Maharashtra,India):DevelopmentandvalidationofHPTLCdensitometricquantificationmethodforgallicacidfromPhyllanthusemblicaLinn.AbstractNo.C-29�,61stIPC(2009).HPTLCofgallicacidinfruitsofPhyllanthusemblicaonsilicagelwithtoluene-ethylacetate-formicacid-methanol15:15:4:1.ThehRfvalueofgallicacidwas40.Quantitativeabsorbancemeasurementat280nm.Themethodwaslinearintherangeof40-240ng/band.Themethodwasreproducibleandsuitableforqualitycontrol.

pharmaceuticalresearch,qualitycontrol,herbal,HPTLC,densitometry �2e

105108 M.SAXENA,K.RAVIKANTH,A.KUMAR*,A.GUPTA,B.SINGH,A.SHARMA(*Phyto-chemistryandAnalyticalLaboratory,RandDCentre,AyurvetLimited,Baddi,H.P.,India,[email protected]):PurificationofAzadirachtaindicaseedcakeanditsimpactinnutritionalandantinutritionalfactors.J.Agric.FoodChem.58,49�9-4944(2010).HPTLCofazadirachtin(1)andsalannin(2)intheseedsofAzadirachtaindicaonsilicagelwithhexane-ethylacetate1:�.Quantitativedeterminationbyabsorbancemeasurementat220nm.ThehRfvaluesof(1)and(2)were18and�0,respectively.Linearitywasbetween50and200ppmforboth(1)and(2).Reco-verywas99.9%for(1)and99.�%for(2).Theintermediateprecisionwas88.5%and87.5%for(1)and(2),respectively(n=�).TheHPTLCandHPLCmethodsgavecomparableresults.

foodanalysis,herbal,HPTLC,quantitativeanalysis,densitometry,comparisonofmethods �2e

105109 M.SCHULZ*,S.MINARIK,C.WIRTH,M.OBERLE(*MerckKGaA,PC-RP-SIL,Frankfur-terStr.250,6429�Darmstadt,Germany):Screeningofunknownplantextractsbyplanarchro-matogaphy.CBS10�,10-12 (2009).HPTLCofplant extractsand standardschlorogenicacid,hyperoside,rutin,quercetinandkaempferol(0.1%inmethanol)onsilicagelinatwin-troughchamberwithethylacetate-formicacid-glacialaceticacid-water100:11:11:27.Detectionbysprayingwithvariousdetectionreagents:1)naturalproductsreagent,evaluationunderUV�66nm,2)anisaldehydereagent,evaluationunderwhitelight,�)diphenyl-2-picrylhydrazylreagent(DPPH),evaluationunderwhitelight,4)rhodamineBreagent,evaluationunderUV�66nm,5)Dragendorffreagent,evaluationunderwhitelight.

herbal,HPTLC,qualitativeidentification �2e

105110 N.SHARMA,U.SHARMA,A.GUPTA,A.SINHA*(*NaturalPlantProductsDivision,Insti-tuteofHimalayanBioresourceTechnology(CSIR),HimachalPradesh,India,[email protected]):Simultaneousdeterminationofepicatechin,syringicacid,quercetin-�-O-galactosideandquercitrin in the leavesofRhododendronspeciesbyusingavalidatedHPTLCmethod. J.FoodComp.Anal.2�,214-219(2010)HPTLCofepicatechin(1),syringicacid(2),quercetin-�-O-galactoside(�),andquercitrin(4)intheleavesofRhododendronspeciesonRP18withme-thanol-5%formicacidinwater1:1.Quantitativedeterminationbyabsorbancemeasurementat290nm.ThehRfvaluesof(1),(2),(�),and(4)were6�,47,28and21,respectively.Linearitywasbetween200and1200ngfor(1),(�)and(4),andbetween200and2400ngfor(2).Theintra-dayandinter-dayprecisions(expressedintermsof%RSD)forcompounds(1)to(4)wereintherangeof0.41-1.�7%and0.67-2.04%,respectively.LODobtainedforcompounds(1)to(4)were20,40,25and25ng/zone,respectivelywhileLOQwere50,115,75and70ng/zone,respectively.Recoveryforallfourcompoundswasintherangeof95.5-98.5%.

herbal,HPTLC,densitometry,quantitativeanalysis �2e

105111 A.N. SHIKOV*, Olga N. POZHARITSKAYA, SvetlanaA. IVANOVA,V.G. MAKAROV,V.P.TIKHONOV,B.GALAMBOSI(*SaintPetersburgInstituteofPharmacy,47/5,Piskarevskiypr.,195067,St.Petersburg,Russia;[email protected]):ImprovedandvalidatedHPTLCmethodfor


quantificationofoenotheinBand itsuse foranalysisofEpilobiumangustifoliumL. J.PlanarChromatogr.2�,70-74(2010).DescriptonofaselectiveandsimpleHPTLCmethodforquanti-ficationofoenotheinBonthebasisofthefreegallicacidandtotalgallicacidcontentafteracidhydrolysis.HPTLCofgallicacidonsilicagelwithbenzene-methanol-aceticacid90:16:8inaglasschamberpreviouslysaturatedwiththemobilephasevaporfor20min.Quantitativedeter-minationbyabsorbancemeasurementat570nmafterderivatizationwith1%ethanoliciron(III)chloridesolution.Averagerecoveryoftheactiveingredientwasintherange95.4-104.6%.Line-aritywasintherangeof440-2200ng/band.Thecorrelationcoefficientrwas0.9991,LOD/LOQwere120/�60ng/band;repeatability(RSD)was�.0%andintermediateprecision1.0%;intradayprecision(RSD,n=6,440-2200ng/band)was�.8to5.2%andinterdayprecision4.�to5.7%.Both,precisionandaccuracy,werewithinacceptablelimitsforroutinedruganalysis(</=15%).

herbal,qualitycontrol,HPTLC,quantitativeanalysis,densitometry �2e

105112 A.SHIKOV*,OlgaPOZHARITSKAYA,SvetlanaIVANOVA,V.MAKAROV,VeraKOSMAN(*Saint Petersburg Institute of Pharmacy, 47/5, Piskarevsky prospect, 195067, St. Petersburg,Russia,[email protected]):AcomparisonbetweenHPLCandHPTLCforthesepara-tion andquantificationofboswellic acids inBoswellia serrata extracts.CBS104, 2-4 (2010).HPTLCofbeta-boswellicacid(BA),acetyl-beta-boswellicacid(ABA),11-keto-beta-boswellicacid(KBA),andacetyl-11-keto-beta-boswellicacid(AKBA)inBoswelliaserrataextractsonsi-licagelwithn-hexane-ethylacetate-glacialaceticacid16:5:1inatwin-troughchambersatu-ratedfor15min.Quantitativedeterminationbyabsorptionmeasurementat254nmforKBAandAKBA,andat560nmforBAandABAafterderivatizationbymanualdippinginanisaldehydereagent follwedbyheatingat110°Cfor5min.Resultswerecomparedwith results from theHPLCanalysis.ByHPLC(injectionvolume20µL)thelimitsofdetectionforKBAandAKBAwere6-8ng,andforBAandABA60-80ng.ByHPTLC(applicationvolume2µL)thelimitsofdetectionforKBAandAKBAwere150ng,andforBAandABA100ng.Precision(%RSDofboswellicacids)byHPLCwasbetween6and18%andbyHPTLCbelow2%forAKBAandKBA,andaround10%forBAandABAafterderivatization.ComparisonofquantificationofboswellicacidinextractsbyeitherHPLCorHPTLCshowedthatbothmethodsgaveidenticalre-sults.HPTLCisthemethodofchoiceforroutineanalysisbecauseoflowersolventconsumptionandfastanalysistime.

HPTLC �2e

10511� JyotiSHRIVASTAVA*,M.MAHADIK,S.DHANESHWAR(PoonaCollegeofPharmacy,Dept.foPharmaceuticsChem.,BharatiVidyapeethUniv.,Mah.,India):ValidatedHPTLCmethodde-velopment for simultaneous quantitation of thiocolchicoside and diclofenac in bulk drug andformulation.InternationalSeminaronHerbalDrugResearch,PN-017(2009).HPTLCofthio-colchicosideanddiclofenacsodiumonsilicagelwithtoluene-acetone-methanol-formicacid500:200:200:1.Quantitativedeterminationbyabsorbancemeasurementat280nm.Themethodwas linear in the rangeof160-800ng/band (thiocolchicoside)and1000-5000ng/band (diclo-fenacsodium).Therecoverywasintherangeof99.2-100.9forbothcompounds.

pharmaceutical,research,densitometry,quantitativeanalysis,HPTLC �2a,17c

105114 S.SHUKLA*,A.SHUKLA,S.PANDYA(*Indukaka IpcowalaCollegeofPharmacy,NewVVNagar,Gujarat,India):AvalidatedHPTLCmethodforthequantificationofursolicacidandluteolininLippianodifloraRich.AbstractNo.C-454,61stIPC(2009).HPTLCofursolicacidandluteolinfromaerialpartsofLippianodifloraonsilicagelwithtoluene-ethylacetate-for-micacid70:�0:�.ThehRfvalueofluteolinwas�4andofursolicacid85.Densitometricanalysisat254nmforluteolinandat5�0nmforursolicacidafterderivatizationwithnaturalproducts


reagentfollowedbyPEGreagent.Therecoveryofbothmarkercomponentswasintherangeof98.6-100.5%.


105115 Y.SUBUDHA*,K.VARSHNEY,S.EDWIN,S.AHMED(*B.R.NahataCollegeofPharmacy,Mandsaur,M.P.,India):EstimationofcurcumininamarketedherbalproductRheumax(Herba-julesrumatis)usingHPTLC.AbstractNo.C-291,61stIPC(2009).QuantitativedeterminationofcurcumininthemarketedherbalproductRheumax(Herbajulesrumatis)containingextractsofCurcumalonga,Boswelliaserrata,TinosporacordifoliaandVitexnegundobyTLConsilicagelwithchloroform-methanol�7:�.ThehRf valueofcurcuminwas59.Quantitativeabsor-bancemeasurementat4�0nm.Thelinearregressionbetween100and500ng/zoneshowedr2of0.99984andRSDof1.58%forcurcumin.

pharmaceuticalresearch,qualitycontrol,herbal,densitometry,quantitativeanalysis,HPTLC �2e

105116 N.SURYAVAMSA*,S.MANIMARAN,G.ARUN,K.NITHYA,S.DHANBAL,T.PRAVEEN(*J.S.S.CollegeofPharmacy,DeptofPhytopharma&Phytomedicine,TIFACOREHD,Myso-re,T.N.,India):ValidationanddeterminationofpossiblecatechinspresentinCamelliasinensiscollectedfromdifferentplaceofIndia.InternationalSeminaronHerbalDrugResearch,PN-024(2009).HPTLCofcatechinsandepicatechininleavesoftea(Camelliasinensisonsilicagelwithtoluene-ethylacetate-formicacid7:5:1.Quantitativedeterminationbyabsorbancemeasure-mentat254nm.Theleaveswerefoundtocontain10-24%oftotalpolyphenols.

pharmaceuticalresearch,herbal,densitometry,comparisonofmethods,HPTLC �2e,7

105117 S.R.TAMBE*,R.H.SHINDE,L.R.GUPTA,V.PAREEK,S.B.BHALERAO(*MahatmaGan-dhiVidyamandir‘sPharmacyCollege,Panchavati,Mumbai,AgraRoad,Nashik42200�,Maha-rashtra, India; [email protected]):DevelopmentofLLEandSPEproceduresanditsapplicationsfordeterminationofolmesartaninhumanplasmausingRP-HPLCandHPTLC.J.Liq.Chromatogr.Relat.Technol.��,42�-4�0(2010).HPTLCofolmesartanandzidovidineonsilicagelwithethylacetate-methanol-aceticacid160:40:1inatwin-troughchambersatu-ratedfor10min.Quantitativedeterminationbydensitometricscanningat269nm.Thelinearityrangewas80-600ng/zone.LOQwas80ng/zone,thecorrelationcoefficient0.9900and0.9820.Recoverywas90.1and79.6%.Theaccuracyandprecisionofthemethodweredeterminedbyrepeatability(intra-day)andintermediateprecision(inter-day)forthesetofqualitycontrolsam-ples(low,mid,high)inreplicate.Theresultsrevealedexcellentintra-andinter-dayaccuracyandprecisionofthemethod,whichwaswithintheacceptablelimit(accuracy(%RE)11.89and6.76(low),2.5�and�.8�(mid),and0.65and7.14(high); inter-dayprecision(CV):�.29and�.00(low),1.02and1.51(mid),and1.11and0.69(high);intra-dayprecision:2.59and2.86(low),1.07and1.1�(mid),and1.04and0.72(high)-afterLLEandSPE,respectively).


105118 T.THOMAS*,R.JAYAPRAKASAM,M.GANDHIMATHI,T.RAVI(*CollegeofPharma.,SriRamakrishnaInstituteofParaMedicalSciences,S.N.Raod,Coimbatore,T.N.,India):HPTLCfingerprintingandevaluationofantioxidantactivityofextractsofCyperusrotundusLinnrhizo-mes.InternationalSeminaronHerbalDrugResearch,PN-051(2009).HPTLCoffourdifferentextractsofrhizomesofCyperusrotundusonsilicagelwithchloroform-benzene1:1.Evaluationunder254nm.Detectionbytreatmentwithethanolicpotassiumhydroxideandevaluationofthefingerprintunderdaylight.

herbal,HPTLC �2e

CAMAGBIBLIOGRAPHYSERVICE No.105��

105119 M.TOMCZYK*,A.BAZYLKO,A.STASZEWSKA(*DepartmentofPharmacognosy,Facul-ty of Pharmacy,Medical University of Bialystok, Bialystok, Poland, [email protected]):DeterminationofpolyphenolicsinextractsofPotentillaspeciesbyhigh-performancethin-layerchromatographyphotodensitometrymethod.Phytochem.Anal.21,174-179(2010).HPTLCoftiliroside(1),methylbrevifolincarboxylate(2),andellagicacid(�)intheaerialpartsofPotentillaspeciesonsilicagelwithtoluene-ethylformate-formicacid6:4:1.Quantitativedeterminati-onbyabsorbancemeasurementat�20nmfor(1),287nmfor(2),and280nmfor(�).ThehRfvaluesof(1),(2)and(�)were9,1�,and20,respectively.Linearitywasbetween50and500ng/zonefor(1),50and520ng/zonefor(2),and52and500ng/zonefor(�).Theintra-andinter-dayprecisions(expressedintermsofCV%)wereobservedintheranges2.5-9.0%and2.4-11.1%,respectively.LODfor(1)to(�)were5,10and�4ng/zone,respectively,whileLOQwere27,17and44ng/zone,respectively.Theaveragerecoveryof(1),(2)and(�)was101.1,82.2and94.0%,respectively.

herbal,HPTLC,densitometry,quantitativeanalysis �2e

105120 AnnaW.SOBANSKA*,E.BRZEZINSKA(*DepartmentofAnalyticalChemistry,MedicalUni-versityofLodz,ul.Muszynskiego1,90-151Lodz,Poland;[email protected]):RapidHPTLCquantificationofp-aminobenzoicacidincomplexpharmaceuticalpreparations.J.PlanarChro-matogr.2�,141-147(2010).HPTLCofp-aminobenzoicacidonsilicagel(prewashedwiththemobilephase)withdiethylether-cyclohexane5:1inanunsaturatedchamber.Quantitativedeter-minationbyabsorbancemeasurementat270nm.Detectionbyautomaticsprayingthefollowingreagent solutions: 0.1mol/L sodiumnitrite solution, 0.1mol/Lhydrochloric acid, 0.0�mol/Lethanolic8-hydroxyquinolinesolution,and10%sodiumhydroxidesolution.Foreachsolution2sprayingcycleswereapplied.Afterdryingquantitativedeterminationbydensitometricscan-ningat500nm.Measuredbeforederivatizationat270nmcalibrationofp-aminobenzoicacidwasperfomedbetween50-1600ngperspot.LODwas170ng/spot(calculated)and50ng/spot(estimatedvisually).LOQ(calculated)was510ng/spot.Thecorrelationcoefficientrwas0.99�6(peakarea)and0.9866(peakheight);thelinearrangewas200-1800ng/spot.Measuredat500nmafterderivatization,LODwas290ng/spot(calculated)and50ng/spot(estimatedvisually).LOQ(calculated)was870ng/spotand200ngperspot(estimatedvisually).Thecorrelationcoef-ficientrwas0.9815and0.95�8.


105121 D.YENICELI,D.DOGRUKOL-AK*(*AnadoluUniversity,FacultyofPharmacy,DepartmentofAnalyticalChemistry,26470Eskisehir,Turkey;[email protected]):Avalidated thin-layerchromatographic method for analysis of bupropion hydrochloride in a pharmaceutical dosageform.J.PlanarChromatogr.2�,212-218(2010).TLCofbupropiononsilicagelwithethanol-chloroform-glacialaceticacid�0:10:1.ThehRfvaluewas56.Quantitativedeterminationbydensitometryat254nm.Linearitywasintherange200-1000ng/band(viapeakarea).Thelimitsofdetectionandquantitationwere11and�5ngperband,respectively.Theintra-dayrepeatabilityofthemethodwasaround1-2%RSD.Recoverywasbetween102.1and104.6%andbetween97.2and102.2%forquality-controlstandardsandforbupropionhydrochloride,respectively.


33. Inorganic substances

105122 S.S.BOZKURT,I.K.CAVDAR,H.M.KURTBAY,M.MERDIVAN*(*DokuzEylulUniversity,FacultyofArtsandSciences,ChemistryDepartment,�5160BucaIzmir;[email protected]):Determinationoftracemetalionsusingporphyrinsaschelatingagentsbyhigh-perfor-mance thin-layerchromatography-densitometry. J.Liq.Chromatogr.Relat.Technol.��,748-760 (2010). HPTLC of mercury(II), copper(II), nickel(II), cadmium(II), mercury(II), lead(II),cobalt(II),manganese(II),palladium(II),platinum(II),andzinc(II)asporphyrincomplexesusingnewlysynthesizedtetra-(bromo-4-hydroxyphenyl)porphyrin(TBHPP),tetra-(4-phenoxyphenyl)


porphyrin(TPPP)andtetra-p-chloromethylphenylporphyrin(CMPP)onsilicagelwithacetone-chloroform1:4forTBHPPandTPPPanddichloromethane-chloroform-hexane1:1:�forCMPP.ThehRfvaluesweredeterminedforthemetal-TBHPP,-TPPP,and-CMPPchelates.Detectionwasperformedbyabsorbancemeasurementat420nm.Thelinearrangewas�.6-60,�.6-�0,1.2-�0,0.6-�0,0.6-�0,2.4-60ng/µL,LODwas0.90,0.92,0.�6,0.19,0.16,and0.41ng/µL,andLOQwas�.01,�.06,1.11,0.54,0.54,0.54,and1.�6ng/µL. Intermediateprecision (RSD%,n=5)(6ng/µL)was�.55,4.25,�.20,�.85,2.25,0.45%and the regressioncoefficientwas0.9908,0.9904,0.9944,0.9961,0.9972,and0.9942.ThehRfvalueswere68,16,8�,42,67,and76forHg-TBHPP,Zn-TBHPP,Cu-TBHPP,Co-TBHPP,Hg-TBHPP,andCu-CMPP,respectively.

HPTLC,densitometry,quantitativeanalysis ��a

10512� A.MOHAMMAD*,A.MOHEMAN(*AnalyticalResearchLaboratory,DepartmentofAppliedChemistry,FacultyofEngineeringandTechnology,AligarhMuslimUniversity;Aligarh202002,India;[email protected]):Adsorption of zinc(II) and cadmium(II) on soil layers inTLC in thepresenceof surfactant-containingmobile phases. J. PlanarChromatogr. 2�, 28-�4(2010).Plateswithlayersofspecifiedsoilwerepreparedbycoating20cm×�.5cmglassplatesbyasoilslurry(soil-to-waterratio1:2)togivelayers0.5mmthick.Theplateswerethendriedinairatroomtemperature(�0±2°C).TLCofZn(II)andCd(II)nitrateonsoilplateswith1.0Mmagnesiumchloridesolutionin9.9mMCTAB(cetaltrimethylammoniumbromide)astheopti-mummobilephase(of�5mobilephases).Detectionbysprayingwithdithizonesolution(0.1%incarbontetrachloride)toobtaindarkpinkandbrickredspotsofcomplexes.Thesmallestdetec-tableamountsofZn(II)andCd(II)onsoillayerswere0.69and1.0µg,respectively,fordistilledwaterand9.0mMCTABsolution,and1.2and2.5µg,respectively,fortheoptimummobilephase.

qualitativeidentification ��a

105124 A.MOHAMMAD*,S.HENA,A.MOHEMAN(*AnalyticalResearchLaboratory,DepartmentofAppliedChemistry,FacultyofEngineeringandTechnology,AligarhMuslimUniversity,Ali-garh-202002,India;[email protected]):MicellarTLCofinorganicions:Simultane-ousseparationoflead(II),zinc(II),andcobalt(II)andspectrophotometricestimationofzinc(II).J. Planar Chromatogr. 2�, 162-165 (2010).TLC of iron(III), copper(II), nickel(II), cobalt(II),cadmium(II),zinc(II),silver((I),lead(II),bismuth(III),mercury(II),titanium(IV),manganese(II),andchromium(VI)onsilicagelwith0.02,0.1,0.2,and1.0Maqueoussodiumdodecylsulfate(SDS)(M1),0.2MSDS+0.04Mtartaricacid(9:1,1:1,and1:9)(M2);0.2MSDS+0.08Mtar-taricacid(9:1,1:1,and1:9)(M�);0.2MSDS+0.1Mtartaricacid(9:1,1:1,and1:9)(M4);0.2MSDS+0.08Mcitricacid(1:1)(M5);0.2MSDS+0.08Mformicacid(1:1)(M6);0.2MSDS+0.08Maceticacid(1:1)(M7);0.2MSDS+0.08Moxalicacid(1:1)(M8).Detectionreagentsusedwere:0.5%dithizoneincarbontetrachloride,1.0%aqueouspotassiumferrocyanide,1.0%ethanolicdimethylglyoxim,1:12.0Maqueoussodiumhydroxidein�0%hydrogenperoxide,andamethanolicsilvernitratesolution.Quantitativeanalysisofzinc(II)byspectrophotometryafterextraction.Thedetectionlimitswere0.85,0.05,and1.5µgrespectivelyforlead(II),zinc(II),andcobalt(II).Thein-situdetectionofcationswasmoresensitivethandetectioninsolution.

qualitativeidentification ��a

105125 P.A.M.NAJAR*,R.N.CHOUHAN,M.T.NIMJE,K.V.R.RAO(*JawaharlalNehruAluminiumResearchDevelopmentandDesignCenter,Wadi,AmaravatiRoad,Nagpur-44002�,India;[email protected]):Quantitativeanalysisofprimaryaluminiumbydensitometricthin-layerchro-matography.J.PlanarChromatogr.2�,156-161(2010).TLCofaluminium,siliconandirononmicrocrystallinecellulose,onsilicagelandcombinationswithaluminiumoxide.Mobilephasesusedwere1:1,7:�,9:1,and8:2mixturesof10%aqueoussolutionsofsodiumchloride,potas-sium chloride, potassium bromide, and sodium formate with 10 % aqueous formic acid, and1:8 mixtures of 1 % aqueous sodium molybdate with � %, 4 %, and 5 % hydrochloric acid.


Aluminium(III)wasdetectedbysprayingwith0.05%aqueousaluminon(triammoniumaurintricarboxylate).Aqueouspotassiumferrocyanidein10%aqueoussodiumformatesolutionwasusedforthevisualizationofiron(II).Siliconwasdetectedbysprayingwith1%sodiummolyb-dateatpH0.80-0.92.Quantitativedeterminationbyscanningdensitometryat402nmforsilicon,at528nmforaluminiumandat620nmforiron.LODwas7-10ppmforaluminium(III),2-5ppmforiron(II),and4-6ppmforsilicon(IV).

densitometry,quantitativeanalysis ��a

38. Chiral separation

105126 RaviBUSHAN*,CharuAGARWAL(*DepartmentofChemistry,IndianIstituteofTechnology,Roorkee-247667,India;[email protected]):ResolutionofbetablockerenantiomersbyTLCwithvancomycinasimpregnatingagentoraschiralmobilphaseadditive.J.PlanarChromatogr.2�,7-1�(2010).TLCoftheenantiomersofracemicatenolol,metoprolol,propanolol,andlabeta-lolonsilicagelusingvancomycinaschiralimpregnatingagentoraschiralphaseadditive.Withvancomycinasimpregnatingagent,resolutionoftheenantiomersofatenolol,metoprolol,propa-nolol,andlabetalolwasachievedwithacetonitrile-methanol-water-dichloromethane7:1:1:1,acetonitrile-methanol-water6:1:1,acetonitrile-methanol-water-dichloromethane-glacialaceticacid14:2:2:2:1,andacetonitrile-methanol-water15:1:1.Withvancomycinasmobilepha-seadditive,resolutionoftheenantiomersofmetoprolol,propanolol,andlabetalolwasachievedusingacetonitrile-methanol-0.56mMaqueousvancomycin(pH5.5)15:1:2,andacetonitrile-methanol - 0.56 mM aqueous vancomycin (pH 5.5) - dichloromethane 18:2:�:2, respectively.Chromatogramsweredevelopedinglasschamberspreviouslyequilibratedwiththemobilephaseat16+/-2°Cfor10-15min.Detectionbyexposuretoiodinevapor.Thedetectionlimitswere1.�,1.2,1.5.and1.4µgforeachenantiomerofatenolol,metoprolol,propanolol,andlabetalol,respectively.

pharmaceuticalresearch,qualitycontrol,qualitativeidentification �8


International Symposium for High-Performance Thin-Layer Chromatography

BASEL, Switzerland 06 – 08 July 2011

We would like to inform you about an outstanding event taking place, the

This symposium will comprehensively inform scientists about the immense potential of HPTLC and its latest developments. Be up-to-date with the state-of-the-art. HPTLC is increasingly seen as a rational method in this age of ultra-rapid separations and an exchange of knowledge is foremost in sustaining this position. The scientific program will feature tutorials, invited keynote speakers, selected submitted lectures, panel discus-sions, and poster presentations. Contributions are invited from all areas of planar chromatography.

Deadlines

• Abstract submission (oral and poster): 1 March 2011

• Final registration: 30 May 2011

For abstract submission and electronic registration see www.hptlc.com. An abstract template (guideline) is available for download. After the abstract submission deadline the final program will be announced at this webpage, about the middle of March.

Tentative program The program will run from Wednesday 6th 9:15 (registration starts at 8:00) until Friday 8th 15:30.

Wednesday 9:15-10:30 two parallel tutorials on HPTLC analysis of herbal and medicinal plants (Dr. Eike Reich) and Hyphenations in HPTLC (Dr. Gertrud Morlock) are planned prior to the formal opening of the symposium at 11:00. Dedicated panel discus-sions between sessions will provide a focus and forum for the exchange of information on the main themes of the sympo-sium. Presentations will end by 13:00 on Friday 8th to allow for lunch and visit to the Novartis campus in the afternoon.

Among others, invited speakers include:

Fundamentals: Prof. Dr. Colin Poole, USA

Analysis of medicinal plants: Prof. Dr. Ikhlas Khan, USA, Prof. Dr. Zheng-Tao Wang, China

Analysis of phytopharmaceuticals and herbal products: Dr. Wanchai De-Eknamkul, Thailand, Dr. J. Madhusudana Rao, India

Analysis of cosmetics/active ingredients: Prof. Dr. Ingo Schellenberg, Germany

Food analysis: Prof. Dr. Mario Vega, Chile

Detection with bioassays: Dr. Irena Choma, Poland

HPTLC-MS: Dr. Gary Van Berkel, USA

Miniaturization: Prof. Dr. Michael Brett/Steven Jim, Canada

Call for papers

Exhibition There will be a manufacturers’ exhibition in the Congress Center in parallel with the seminar program.

Social events Wednesday evening: Welcome cocktail at Ramada hotel Thursday afternoon: Manufacturers’ cocktail Thursday evening: Official symposium dinner: Rhine cruise with a marvellous view of Basle Friday afternoon: Visit to the new Novartis campus

Location Wednesday/Thursday Congress Center Messeplatz 21 4005 Basel www.congress.ch

Friday: Novartis Pharma AG Auditorium Fabrikstr. 6 4002 Basel www.novartis.ch

Special accommodation rates are available for the Symposium: RAMADA PLAZA Messeplatz 12 4005 Basel www.ramada.de/basel

Fees The participation fee includes the full scientific program, lunches, coffee breaks, and all social events.

• Industrial 500 €

• Academic 400 €

• Students 200 €

Reduction of 100 € with ISPS or CCCM membership.

Many returning faces from previous symposia and new faces are cordially invited. We are looking forward to welcoming you at the conference.

Further information [email protected] for general information [email protected] for scientific matters [email protected] for registration concerns

8 CBS 105

Know CAMAG

One of CAMAG’s goals is to provide worldwide in a variety of venues, a continuous flow of information on the state of up to date knowledge of contempo-rary planar chromatography (see editorial CBS 104). Through one-day workshops at universities, techni-cal training schools, official governmental institutes, and at private companies through in-house training of laboratory personnel, this goal is a reality.

1. The beginning: How we got started!

Lectures and practical sessions on analytical chemi-stry for our students concentrated on Separation Sciences, i.e. instrumental bio-analysis, protein and carbohydrate analysis, hyphenating techniques, etc. Meanwhile, contemporary TLC/HPTLC has enhanced the scope of our “HPLC oriented” labo-ratories considerably.

A related institute offered us their CAMAG equip-ment, which fortunately for us had been mistakenly considered to be non-functional. CAMAG provided generous and efficient support. First they checked the TLC Scanner free of charge, and found it fully functional. Then they instructed our personnel in the operation of the equipment and the use of the software. For this help we are thankful to Dr. Nat-sias, Ms Werther, Dr. Zieloff and Mr. Nachtwey, all from CAMAG Berlin.

Prof. Gey, Ms Werther, Ms Dr. Gey, Dr. Zieloff (from left) Practical session11 12

In Germany, CAMAG Berlin has held such seminars for over ten years, now extended into Austria and Benelux. The USA has a similar program and India has an even more aggressive program. In all other countries these are organized by CAMAG Switzer-land. If you are interested, contact [email protected] or telephone +41 61 4673434.

2. Workshop on contemporary TLC/HPTLC 17./18. May 2010 in Zittau

Very soon our cooperation with CAMAG led to the plan to organize a workshop on TLC/HPTLC for students and co-workers from our Faculty for Mathematics & Natural Sciences. There were 30 participants, considered a very good turnout, since all regular lectures and practical sessions ran on in parallel. Dr. Zieloff and Ms Werther demonstrated their outstanding knowledge in their lectures and experimental sessions. The participants were given the opportunity to optimize separations themselves. All results were critically evaluated and discussed. Meanwhile the workshop has now been incorpo-rated in our teaching program as a fixed element.

All in all, the event was considered a great success. Once again a big thank you to CAMAG!

About one such seminars, recently held by CAMAG Berlin, Professor Manfred Gey of the University of Applied Sciences Zittau/Görlitz reports:

CBS 105 9

15

13

continuation from page 7

Further information is available from the authors on request.

Contact: Dr. Walter H. Weber, Zweckverband Landeswasserver-sorgung, Betriebs- und Forschungslaboratorium, Am Spitzigen Berg 1, 89129 Langenau, Germany, [email protected]

[1] W.H. Weber et al. Vom Wasser 105 (1) (2007) 7

[2] W.H. Weber et al. Vom Wasser 107 (4) (2009) 16

[3] A. Müller et al. Rapid Commun Mass Spectrom 24 (2010) 659

HPTLC/AMD chromatogram of SPE extracts from various secondary effluents and of the standard mixture 1H-benzo-triazole/tolyltriazole

25-step AMD 2 gradient for the screening of water samples

HPTLC/AMD multi-wavelength scan of an extract from se-condary effluents (a), Nano LC/QTOF-MS chromatogram of the extracted zone, peak A: 1H-benzotriazole; peak B: tolyl-triazole (b)

For the identification of the two unknown analytes a substance library (approx. 300 entries) created at the laboratory for operation control and research of the LW was used, which features molecular formula and exact mass of potential environmental contami-nants described in literature. The unknown analytes were identified as 1H-benzotriazole and a mixture of 4-methyl-1H-benzotriazole and 5-methyl-1H-benzotriazole.

Benzotriazoles are ubiquitous in the aquatic envi-ronment because they are used in a broad range of applications: in coatings and paints, as anticorrosive in copper and copper alloys, in coolants and lubri-cants for engines, as silver protection in detergents, as antifreeze and as aircraft de-icing fluid.

In 14 % of ground water samples (n = 74) 1H-benzotriazole was detected at a maximum of 173 ng/L and in 18 % of samples tolyltriazoles were detected at a maximum of 75 ng/L. Concentrations in the

14

river Danube and in some of its feeding rivers in the Ulm area were between 100 and 500 ng/L. In sec-ondary effluents, the main source of benzotriazoles in the environment, concentrations of 1H-benzot-riazole and tolyltriazoles can even exceed 10 µg/L. Comparison of water sampled from wastewater treatment plants, surface water and ground water showed that the ratio of the investigated substances shifted towards 1H-benzotriazole.

10 CBS 105

16


Optimization of an AMD 2 method for determination of stratum corneum lipids

Prof. Dr. Ingo Schellenberg and Dr. Kathrin Kabrodt

The workgroup of Prof. Schellenberg at the Institute of Bioanalytical Sciences (IBAS) is engaged in the isolation and production of plant extracts, which are used in foodstuffs, in nutriceuticals, as well as in cosmetic and pharmaceutical products. Further core competencies are the encapsulation of bioactive in-gredients, lipids and flavors, flavors also investigated as to their importance for sensory quality. Parts of these investigations were done by Ms Julia Lüttich in the course of her Bachelor thesis.

IntroductionLipids are substances that are soluble in organic solvents due to their non polar properties. In stra-tum corneum, which represents the outer layer of skin, lipids play a key role in maintaining the barrier function of the skin, i.e. protection. Predominantly lipids of stratum corneum are ceramides, fatty acids and cholesterol. Ceramides consist of long-chain hydroxylated amine base called sphingoid, which are amide-linked to fatty acids. By various combina-tions of fatty acids and base types there are differ-ent ceramide groups. In human stratum corneum 9 ceramide groups have been identified up to now.

An optimized AMD2 separation of various li-pid standards that are found in native stratum corneum is presented below. The AMD system was already successfully used for these analy-ses (see CBS 77, 90 and 93). However, the de-

scribed methods were rather time and solvent consuming.

Standard solutions

Standard substances Abbre-viation

Supplier Sample amount

(mg)

Concentration of standard

solution (mg/mL)

Ceramide NS NS Sederma* 14,0 0,35

Ceramide NP NP Evonik* 4,8 0,12

Ceramide AS AS Sigma-Aldrich 4,8 0,12

Ceramide AP AP Evonik* 14,0 0,35

Cholesteryl-3-sulfate C3S Sigma-Aldrich 8,0 0,20

Cholesterol C Sigma-Aldrich 5,6 0,14

Cholesteryloleate CO Sigma-Aldrich 4,0 0,10

Glyceryl trioleate GT Sigma-Aldrich 6,0 0,15

Phosphatidylcholine PC Sigma-Aldrich 4,0 0,10

Oleic acid OA Sigma-Aldrich 3,6 0,09

Squalene S Sigma-Aldrich 8,0 0,20

Sphingomyeline SM Sigma-Aldrich 5,6 0,14

*Thanks to Sederma and Evonik for free supply of standards.

Each standard substance was weighted into a 2 mL volumetric flask each and filled up with methanol – chloroform 1:1 (stock solution). 500 µL of each stock solution were diluted to 10 mL with the same solvent (standard solution). 500 µL of each stock solution were put into a 10 mL volumetric flask and filled up with the same solvent (standard solution mix).

Chromatogram layerHPTLC plates 20 x 10 cm, silica gel 60 F254, 0.1 mm for AMD (Merck); pre-washed twice with chloro-form – methanol 2:1 (v/v) and dried at 120 °C in a drying oven for 30 min; storage in an exsiccator

Sample applicationBandwise with Automatic TLC Sampler 4, 13 tracks, band length 8 mm, distance from the side 14 mm, distance from lower edge 8 mm, track distance 13.6 mm (automatically calculated), application volume 5 µL

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CAMAG AMD 2 System (Automated Multiple Development)

AMD is used when the desired resolution is unattainable over the available separation distance by one step isocratic development. The combination of multiple and gradient de-velopment, and the individual pH adjustment of the layer (here just before step 6) leads to a focusing effect of the zones. Peak sharpness and resolution are improved.

In general, AMD is used for mixtures of components with a wide polarity range. But for this AMD application in the field of lipid analysis, especially the unpolar region of the gradient is of interest. From step to step, the polarity change of the solvent (only 2 %) is minor. This way, also compounds of slight polarity differences are separated. The migration distance increments of mostly 7 mm are high compared to 2 to 4 mm in-crements usually. For a baseline separation using 4 mm increments, a reduced substance amount per zone (e.g. by a factor of 4) would be necessary. Additionally, a final migration distance of 40 mm would significantly reduce the gradient time further on.

Phosphatidylcholine and sphingomyeline (at the start) could not be separated with this gradient. However, if one wants to separate both lipids, it is possible to supplement some polar steps at the beginning of the gradient.

Post-chromatographic derivatizationWith the Chromatogram Immersion Device III the plate was immersed in copper(II)sulfate reagent (aqueous solution of 10% copper(II)sulfate and 8% orthophosphoric acid), then dried and heated at 170 °C on the TLC Plate Heater for 8 min.

DensitometryAbsorption measurement at 600 nm by TLC Scan-ner 3 with winCATS software

DocumentationTLC Visualizer under white light (reflection mode)

Results and discussionThe first step of the 8-step gradient (100 % metha-nol) was used to elute all polar substances and to focus them to a sharp line. Steps two to six sepa-rated cholesterol-3-sulfate and the different cera-mides. Step seven (100 % chloroform) was used to separate cholesteryl oleate from glyceryl trioleate. In the last step with n-hexane – toluene 19:1, squalene was separated from cholesteryl oleate. To focus oleic acid, a pre-conditioning of the HPTLC plate with 4 molar acetic acid was necessary before step six was started.

Chromatography8-step gradient in AMD 2 system, pre-conditioning before step 6 with 4 M acetic acid, drying time 2 min, maximum migration distance 64 mm, duration 1.5 h, solvent consumption 60 mL

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SM PC C3S AP AS NP MIX NS OA C GT CO SQ

AMD 2 chromatogram using the 8-step gradient

Densitogram of the standard mixture

Just phosphatidylcholine and sphingomyeline (at the start position) were not separated by the new gradi-ent. This was also not achieved by Bonté et al. [1] with their 26-step gradient with a duration of 6.8 h. Zellmer et al. [2] and Farwanah et al. [3] did not consider these substances in their methods.

A chromatogram comparison of the 17-step gradi-ent by Farwanah et al. [3] and the optimized 8-step gradient shows a comparable resolution despite the reduced number of steps. The gradient time was reduced by one hour (from 2.5 h to 1.5 h) and the derivatization time was reduced from 20 min (at 150 °C) to 8 min (at 170 °C).

Solvent mixture

Further information is available from the authors on request.

[1] F. Bonté et al. J Chromatogr B 664 (1995) 311

[2] S. Zellmer et al. J Chromatogr B 691 (1997) 321

[3] H. Farwanah et al. J Chromatography B 780 (2002) 443

Contact: Dr. Kathrin Kabrodt, Prof. Dr. Ingo Schellenberg, Anhalt University of Applied Sciences, Center of Life Sciences, Institute of Bioanalytical Sciences, Strenzfelder Allee 28, 06406 Bernburg, Germany, [email protected]

Chromatogram comparison of the 17-step gradient by Farwa-nah et al. ([3], left) and the optimized 8-step gradient (right)

At present this method is in the validation process and in vivo-extraction of defined skin zones is opti-mized to separate and quantify the most important skin lipids.

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Determination of additives in plastic foils

Elisabeth Dytkiewitz, Prof. Dr. Wolfgang Schwack

At the Institute of Food Chemistry, University of Hohenheim, Stuttgart, Germany, efficient analyti-cal work is conceptualized and then developed and implemented. In doing so, planar chromatography is often used due to its pragmatism in solving ana-lytical tasks. The following example concerning the analysis of food packaging materials is part of the doctoral thesis of Elisabeth Dytkiewitz.

IntroductionPlastic foils of polyvinyl chloride (PVC) used for pack-aging meat, cheese, fresh vegetables, etc., contain plasticizers and other additives in high percentages. Most of them have in common that they are not chemically bonded to the polymer, making them potentially free for migration into the packaged food. According to the European legislation, migrat-ing constituents must not endanger human health [1]. Therefore migrating studies with food simulat-ing solvents must be performed to check that the composition is compliant. An effect directed analysis (EDA) brings the bioactivity of substances into focus. Using EDA for the analysis of migrating additives was the aim of this study.

After separation by HPTLC, additives present in migrates are solvent-free accessible, making detection with bioluminescent Vibrio fischeri

bacteria practicable. Therefore many samples can be run in parallel, revealing not only the presence of toxicologically relevant compounds but also information as to their strength of effect. In contrast to common mixed mode cuvette tests, HPTLC-EDA enables the detec-tion of both inhibition and enhancement of luminescence. Detected additives can be identified from the same plate by coupling of HPTLC with mass spectrometry via the TLC-MS Interface. MassWorks software delivers exact masses and molecular formulae even from a mass spectrometer of low resolution. Hence, HPTLC-bioactivity-MS enables rapid analysis of plastic migrate components based on their toxicological effects.

Concerning rapid screenings with focus on major components, the TLC-MS Interface is also suitable for the direct extraction of additives from the packaging films followed by online mass spectrometric analysis.

Sample preparationFoil samples (0.8 g) were extracted by 150 mL etha-nol (95 % vol) in a screw capped bottle for 4 h at 60 °C. After rotary evaporation of the solvent, the residues were taken up in 2 mL toluene. The solu-tions were directly used for HPTLC after membrane filtration.

LayerHPTLC plates silica gel 60 F254, Merck, 20 x 10 cm, prewashed by development with methanol, dried for 15 min in a drying oven at 100 °C.

Sample applicationBandwise with ATS4, band length 6 mm, track distance 18 mm, distance from lower plate edge 8 mm, distance from the edges min. 20 mm, ap-plication volumes of sample solutions 5–10 µL.

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ChromatographyIn the ADC2 with isooctane – toluene – diethyl ether – ethyl acetate 8:7:4:1 after chamber saturation for 10 min up to a migration distance of 65 mm. Plate activity was controlled by saturated magnesium chloride solution for 3 min (42 % relative humid-ity). Plate drying was automatically performed for 30 min.

Detection with biological activityUsing the TLC Chromatogram Immersion Device (vertical speed 3 cm/s, immersion time setting 0 s), the plate was dipped in a suspension of Vibrio fischeri bacteria (BioLuminex kit, ChromaDex, Boul-der, USA) and documented with the Bioluminizer (exposure time 50 s) directly after dipping and after 5 and 10 min.

Mass spectrometryTLC-MS Interface (oval elution head 4 x 2 mm) cou-pled to an Agilent 1100 LC/MSD system, operating in positive ESI mode, zone elution with ethanol (95 %vol) at a flow rate of 0.2 mL/min during 10 s. Exact masses were calculated with MassWorks soft-ware (Cerno Bioscience, Danbury, CT, USA).

Results and discussionEthanol (95 %vol) as a fatty food simulating sol-vent was used for extraction of additives from plastic foils [2]. These migrates were directly used for HPTLC, which was performed by automated standardized development in the ADC 2 to ensure repeatable results. After some modification of the mobile phase [3] a good separation was obtained. Detection with bioluminescent bacteria showed numerous substances with inhibiting effects in the samples. Even in a plastic wrap of polyethylene (PE), luminescence inhibiting substances were detected. Remarkably, some of the zones caused an activat-ing effect on the bacteria’s luminescence, a notable advantage given by the previous chromatography. A cuvette test of the whole migrate would not reveal this property.

Separation and detection of 6 packaging foils; inhibition of bioluminescence displayed by dark zones, enhancement of bioluminescence displayed by light zones. Track assignment: blank: ethanol, P1 and P2: commercial PVC cling film, P3: com-mercial PE cling film, P4: foil of a mushrooms’ packaging, P5: packaging foil for meat, and P6: packaging foil for cheese.

For the identification of EDA detected zones, mass spectra were recorded using the TLC-MS Interface. Ethanol was used to elute the compounds into the ion source. With an elution time of just 10 s, the mass spectrum of the sample zone was achieved im-mediately. Sodium adducts are caused by the saline bacteria suspension remaining at the HPTLC plate surface. After calculation by MassWorks software, the signals from a low resolution mass spectrometer were improved. Thus, exact masses and molecular formula were obtained enabling the identification of additives.

HPTLC-ESI/MS spectra of the light zone at hRF 84 of a mushroom packaging foil (P4).

For a rapid screening of additives extractable from packaging foils, the TLC-MS Interface was directly applied for the transfer of migrates from the foil to the MS, i.e. without chromatography. To ac-complish this, the sample foil was placed on the back of a TLC aluminum foil. After tightening the

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elution head, the mass spectrum was recorded within 10 s.

Direct extraction of a PVC plastic wrap by the TLC-MS Interface. The film was placed planar on the back of a TLC aluminum foil.

Using this short elution time, the highly soluble and more concentrated additives could easily be identified by mass spectrometry, protonated molecules being generally detected.

ESI-MS spectrum of compounds directly extractable from a foil by the TLC-MS Interface (P4)

Applying mass spectrometry, bis(2-ethylhexyl) adi-pate could exemplarily be identified with marginal deviations from the theoretical masses.

MS-Signals of

Mass determined

Theoreticalmass

(ppm) Molecular formula

Assigned to

Plastic foil 371,3174 371,3161 -3,4071 C22H43O4 [M+H]+

HPTLC plate

393,2985 393,2981 -1,0691 C22H42O4Na [M+Na]+

763,6077 763,6064 -1,7164 C44H84O8Na [2M+Na]+

Other substances showing inhibiting effects by Vibrio fischeri are not present in this spectrum. The low con-centration of some substances and the short time of just 10 s for direct foil extraction are the reasons for the reduced elution. Of course, substances eluted into the food simulating solvent could be measured more intensely after a longer contact time simulated by an intermediate flow stop; however, this would also reduce sample throughput.

Direct foil extraction is useful for rapid screening, but the ultimate procedure which produces additional infor-mation is chromatography of migrates, obtained after prolonged heat treatment (4 h at 60 °C) and concentra-tion (by a factor of 75). Thus, information is more com-prehensive. Vibrio fischeri is ideal for detecting other components with toxicological potential, not just major components. It is a specific benefit that from the wealth of these migrating compounds just bio-actives can be detected and identified.

Bis(2-ethylhexyl) adipate

[1] Regulation (EC) No. 1935/2004 as amended on 18.06.2009

[2] Attachment 10 for §11 BedarfsgegenständeVO as amended on 23.09.2009

[3] H. Chen, Y. Wang, R. Zhu, Chinese J Chromatogr A 24 (2006) 69

Further information is available from the author on request.

Contact: Prof. Dr. W. Schwack, University of Hohenheim, Institute of Food Chemistry, Garbenstrasse 28, 70599 Stuttgart, [email protected]

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NEW TLC-MS InterfaceWorld wide used for identification and elucidation of unknown substances in research, forensic and environmental fields

Identification of alkaloids oxymatrine, sophoridine and matrine in Sophora flavescens extract*

Cut and milled Sophora flavescens root parts

HPTLC plate after extraction of zones with the TLC-MS Interface Confirmation of oxymatrine, m/z 265 [M+H]+

*Diploma thesis R. VizziniFurther information at: www.camag.com/tlc-ms

september 2010 - uni-giessen.de · september 2010 the versatility of hptlc – from bio analysis to...

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