lekovito bilje i fitoterapija medicinal plants and ... br 3 2018 lekovito bilje i...

680

Lekovitobiljeifitoterapija

MedicinalPlantsandPhytotherapy

681

SADRŽAJ–CONTENTS

LB-P1

INVESTIGATIONOFANTIBACTERIALPOTENTIALOFTHEPHENOLICSDERIVEDFROMCISTUSINCANUSL.BYMEANSOFTLC‐DB

- Magdalena Knaś, Dariusz Szeremeta, Ewa Długosz, Peter Ott, Mieczysław Sajewicz, Teresa Kowalska, Agnes Moricz 685

LB-P2

HEMIJSKISASTAVIANTIOKSIDACIONAAKTIVNOSTEKSTRAKATAIVANJSKOGCVEĆA,GALIUMVERUML.

CHEMICALCOMPOSITIONANDANTIOXIDANTACTIVITYOFGALIUMVERUML.EXTRACTS

- Jovana Bradić, Anica Petković, Marijana Anđić, Ksenija Vučićević, Miroslav Sovrlić, Ana Barjaktarević, Vladimir Jakovljević, Marina Tomović 686

LB-P3

PHYTOCHEMICALSTUDYOFTHEGREEKENDEMICSPECIESINULASUBFLOCCOSARECH.F.(ASTERACEAE)

- Dafni Ioanna Diakaki, Aikaterini Koutsaviti, Efstathia Ioannou, Olga Tzakou 688

LB-P4

VOLATILECONSTITUENTSOFDIFFERENTPLANTPARTSOFJUNIPERUSOXYCEDRUSSSP.MACROCARPA(SIBTH.&SM.)BALL.FROMGREECE

- Nikolaos Armenis, Maria Couladis 689

LB-P5

ANATOMSKAGRAĐAIHEMIJSKAANALIZAETARSKOGULJAPIMPINELLASAXIFRAGAL.(APIACEAE)

ANATOMYANDCHEMICALANALYSISOFESSENTIALOILOFPIMPINELLASAXIFRAGAL.(APIACEAE)

- Milica Drobac, Bojan Zlatković, Nada Kovačević, Violeta Slavkovska 690

682

LB-P6

ANTIMIKROBNAAKTIVNOSTSOKAPLODAARONIJE(ARONIAMELANOCARPA(MICHX.)ELLIOTT)

ANTIMICROBIALACTIVITYOFCHOKEBERRYJUICE(ARONIAMELANOCARPA(MICHX.)ELLIOTT)

- Milica Milutinović, Katarina Šavikin, Tatjana Mihajilov Krstev, Milica Kostić, Bojana Miladinović, Suzana Branković, Dušanka Kitić 692

LB-P7

FITOHEMIJSKAANALIZAIODREĐIVANJESADRŽAJAFLAVONOIDAUALLIUMURSINUML.,ALLIACEAE

PHYTOCHEMICALANALYSISANDFLAVONOIDCONTENTDETERMINATIONINALLIUMURSINUML.,ALLIACEAE

- Haris Nikšić, Lejla Kadić, Kemal Durić, Irma Sijamić, Nadir Jordamović 694

LB-P8

ISPITIVANJEKVALITETABILJNIHDROGAKOJEULAZEUSASTAVMONOKOMPONENTNIHČAJEVA

QUALITYTESTINGOFHERBALDRUGSINCLUDEDINTHECOMPOSITIONOFMONOCOMPONENTTEAS

- Kemal Durić, Minela Guta-Mujezinović, Haris Nikšić, Irma Sijamić, Samija Muratović 696

LB-P9

FITOHEMIJSKOISPITIVANJETRITERPENSKIHSAPONINASAPOTENCIJALNIMFARMAKOLOŠKIMDJELOVANJEMUVRSTAMAPORODICELAMIACEAE

PHYTOCHEMICALINVESTIGATIONOFTRITERPENESAPONINSWITHPOTENTIALPHARMACOLOGICALACTIVITYINPLANTSPECIESBELONGINGTOLAMIACEAEFAMILY

- Nadir Jordamović, Haris Nikšić, Irma Sijamić, Kemal Durić 698

683

LB-P10

ISPITIVANJEMAKROSKOPSKIH,MIKROSKOPSKIHOSOBINAIETARSKOGULJALISTAMATIČNJAKA,MELISSAOFFICINALISL.

INVESTIGATIONOFMACROSCOPICANDMICROSCOPICCHARACTERISTICSANDESSENTIALOILOFTHELEMONBALMLEAF,MELISSAOFFICINALISL.

- Mirjana Marčetić, Nikola Bošković, Danilo Stojanović, Nada Kovačević 700

LB-P11

MASNEKISELINE,STEROLIITRITERPENIMASNIHULJAPLODOVAOSAMTAKSONARODAHERACLEUML.IZJUGOISTOČNEEVROPE

FATTYACIDS,STEROLSANDTRITERPENESOFTHEFRUITFATTYOILSOFEIGHTHERACLEUML.TAXAFROMSOUTHEASTERNEUROPE

- Ljuboš Ušjak, Ivana Sofrenić, Vele Tešević, Milica Drobac, Marjan Niketić, Silvana Petrović 702

LB-P12

ANTIINFLAMATORNIEFEKATEKSTRAKTAMUSKATNEŽALFIJE(SALVIASCLAREAL.)

THEANTI‐INFLAMATORYEFFECTOFTHECLARYSAGEEXTRACT(SALVIASCLAREAL.)

- Milica Kostić, Dušanka Kitić, Milica B. Petrović, Tatjana Jevtović-Stoimenov, Bojana Miladinović, Milica Milutinović, Bojan Zlatković 704

LB-P13

KONTROLAKVALITETAKOREISPRAŠENEKORECIMETANATRŽIŠTUSRBIJE

QUALITYCONTROLOFCINNAMON’SBARKANDPOWDEREDBARKFROMTHESERBIANMARKET

- Katarina Jeremić, Neda Gavarić, Nebojša Kladar, Nemanja Todorović, Nebojša Salaj, Maja Bekut, Biljana Božin 706

684

LB-P14

VAZORELAKSANTNAAKTIVNOSTTERPINOLENA

VASORELAXANTACTIVITYOFTERPINOLENE

- Relja Suručić, Dragana Drakul, Nada Kovačević, Tatjana Kundaković-Vasović 708

LB-P15

APITERAPIJAUMENOPAUZI

APITHERAPYINMENOPAUSE

- Lidija Jerotijević 710

LB-P16

ANTIMIKROBNAIANTIOKSIDATIVNAAKTIVNOSTMETANOLNOGEKSTRAKTAALCHEMILLAVIRIDIFLORAROTHM.(ROSACEAE)

ANTIMICROBIALANDANTIOXIDANTACTIVITYOFMETHANOLEXTRACTOFALCHEMILLAVIRIDIFLORAROTHM.(ROSACEAE)

- Jelena Radović, Marina Milenković, Tatjana Kundaković-Vasović 712

LB-P17

SADRŽAJESTRAGOLAUETARSKOMULJUIINFUZUHERBEBOSILJKA,OCIMUMBASILICUML.

THECONTENTOFESTRAGOLEINESSENTIALOILANDINFUSIONOFBASILHERB,OCIMUMBASILICUML.

- Jelena Arsenijević, Aleksandar Jovanović, Stefan Jevtić, Nada Kovačević, Milica Drobac 714

LB-P18

ISPITIVANJETRITERPENAUDIHLORMETANSKIMEKSTRAKTIMA28VRSTARODAHIERACIUML.(ASTERACEAE)SABALKANSKOGPOLUOSTRVA

INVESTIGATIONOFTRITERPENESINTHEDICHLOROMETHANEEXTRACTSOF28HIERACIUML.SPECIES(ASTERACEAE)FROMTHEBALKANPENINSULA

- Violeta Milutinović, Marjan Niketić, Miloš Petković, Silvana Petrović 716

685

Arh.farm 2018;68: 685 LB-P1

INVESTIGATIONOFANTIBACTERIALPOTENTIALOFTHE

PHENOLICSDERIVEDFROMCISTUSINCANUSL.BYMEANSOFTLC‐DB

MagdalenaKnaś1,DariuszSzeremeta1,EwaDługosz2,PeterOtt3,

MieczysławSajewicz1,TeresaKowalska1,AgnesMoricz3

1InstituteofChemistry,UniversityofSilesia(Poland),2JanDlugoszUniversityinCzestochowa(Poland),3HungarianAcademyofSciences(Hungary)

Thin‐layer chromatography (TLC) combined with direct bioautography (DB)

(i.e.,TLC‐DB) isaquickandeffectivemethod forpreliminaryassessmentofbiologicalactivityofthemedicinalplantextracts.CistusincanusL.(theCistaceaefamily)ispopularin eastern parts of theMediterranean basin and in theMiddle East, and it has beenknown for centuries as an importantmedicinal herb. Antimicrobial activity of theC.incanusextractsisattributedmainlytothephenolicscontainedinthisplant.Themainobjective of this study was to obtain the TLC fingerprint of fraction I (supposed tocontain flavonoid aglycons alone) derived by selective multi‐step extraction of thephenolicsfromtheplantsample,andthentoscreenitbymeansofTLC‐DBinthesearchforthefingerprintzoneswithantibacterialactivity.

The main analytical technique: thin layer chromatography hyphenated withdirectbioautography(TLC‐DB).Thesupplementarytechnique:HPLC‐DAD‐ESI‐MS.

The antibacterially active zones were found by means of TLC‐DB and provedinhibitingboth,theGrampositiveB.subtilisandtheGramnegativeA.fischeribacterialstrains.Thenanattemptwasundertakentoidentifyatleastsomephenolicsfromactivezones and it was revealed that fraction I contained not only flavonoid aglycons, butsurprisingly enough flavonoid glycosides as well. Among the identified antibacterialcomponents,apigenin,kaempferideandacylatedkaempferolglycosideswerefound.

Onceagain,thin‐layerchromatographywithdirectbioautography(TLC‐DB)hasprovedefficient in screeningextractsderived frommedicinalplants in thesearch fornaturalantibioticsofbotanicalorigin.

686

Arh.farm 2018;68: 686-687 LB-P2

HEMIJSKISASTAVIANTIOKSIDACIONAAKTIVNOSTEKSTRAKATA

IVANJSKOGCVEĆA,GALIUMVERUML.

JovanaBradić1,AnicaPetković1,MarijanaAnđić1,KsenijaVučićević1,MiroslavSovrlić1,AnaBarjaktarević1,

VladimirJakovljević2,3,MarinaTomović1

1Katedrazafarmaciju,UniverzitetuKragujevcu‐Fakultetmedicinskihnauka,2Katedrazafiziologiju,UniverzitetuKragujevcu‐Fakultetmedicinskihnauka(Srbija),3Katedrazapatologiju,PrvimoskovskidržavniUniverzitetI.M.

Sečenov(Rusija)

Ivanjsko cveće (Galium verum) je višegodišnja zeljasta biljka iz roda Galium,

poznata od davnina po terapijskom dejstvu. Međutim njen hemijski sastav, kao ibiološkaaktivnostnisuupotpunostirazjašnjeni.Stogajeciljnašegistraživanjabiodaseutvrdiukupansadržajfenolaiflavonoidaacetonskogivodenogekstraktaivanjskogcveća,kaoiprocenanjihoveantioksidacioneaktivnosti.

Acetonski i vodeni ekstrakti su zasebno pripremljeni ekstrakcijomnadzemnogdela (150 g) biljke sa 500ml odgovarajućeg rastvarača ekstrakcijom pod refluksom.Ukupan sadržaj fenola određen je metodom Folin‐Ciocalteu i izražen kao mgekvivalenta galne kiseline po gramu ekstrakta (mg GAE /g ekstrakta). Procenjen je iukupansadržajflavonoida,arezultatjeizraženkaomgekvivalentakvercetinanagramekstrakta (mg kvercetina/g ekstrakta). In vitro antioksidaciona aktivnost je testiranaspektrofotometrijski pomoću DPPH (1,1‐difenil‐2‐pikrilhidrazil) testa. SposobnostneutralisanjaDPPHradikalajeizraženakaoIC50vrednost.

Naši rezultati su u skladu sa rezultatima drugih studija i pokazali su da je daukupansadržajfenolaacetonskogekstraktaivanjskogcvećaiznosio101,58±7,96mgGAE/gekstrakta.Ovavrednost jenižaodvrednosti za vodeni ekstrakt, čiji je sadržajiznosio127,57 ± 11,25 mg GAE/g ekstrakta. Ukupan sadržaj flavonoida bio je 30,51±3,37 i 61,24 ± 6,11mg kvercetina/g acetonskog i vodenog ekstrakta, redom. Poredtoga, ispitivani ekstrakti su smanjiliDPPHuz IC5011,65±1,16μg/ml za acetonski i9,63±0,32μg/mlzavodeniekstrakt.

Naširezultatisupokazalidaobaekstraktaivanjskogcvećapredstavljajuvredanizvorfenolaiflavonoidaiukazalinaracionalnuosnovuzatradicionalnuupotrebuovebiljnevrsteuublažavanjuoksidativnogstresa.Ovastudijamožebitipolaznaosnovazadalja istraživanja koja bi u potpunosti ispitala hemijski sastav, kao i farmakološkeefekteivanjskogcveća.

687

CHEMICALCOMPOSITIONANDANTIOXIDANTACTIVITYOFGALIUMVERUML.EXTRACTS

JovanaBradić1,AnicaPetković1,MarijanaAndjić1,

KsenijaVučićević1,MiroslavSovrlić1,AnaBarjaktarević1,VladimirJakovljević2,3,MarinaTomović1

1DepartmentofPharmacy,UniversityofKragujevac‐FacultyofMedicalSciences,2DepartmentofPhysiology,UniversityofKragujevac‐Facultyof

MedicalSciences(Serbia),3DepartmentofHumanPathology,1stMoscowStateMedical,UniversityIMSechenov(Russia)

GaliumverumL.isaperennialherbaceousplantbelongingtothegenusGalium,renowned for its therapeutic potential since ancient times. However, its chemicalcompositionaswell asbiological activitieshaven’tbeen fully clarified.Therefore, theaim of our study was to determine the total content of phenols and flavonoids ofacetone and aqueous extracts of G. verum as well as to estimate their antioxidantactivity.

Aqueous and acetone extractswere prepared separately extracting aerial part(150 g)with 500ml of appropriate solvent by heat reflux extraction. Total phenoliccontentwasdeterminedbyFolin‐Ciocalteu'smethodandexpressedasmggallic acidequivalents per gramof extract (mgGAE/g extract). The total flavonoid contentwasestimatedaswellandresultwasexpressedasmgofquercetinequivalentpergramofextract (mg of quercetin/g of extract). In vitro antioxidant activity was evaluatedspectrophotometrically by DPPH (1,1‐diphenyl‐2‐picrylhydrazyl) assay. The DPPHscavengingabilitywasexpressedasIC50value.

Our results are in accordancewith the results of other studies, and illustratedthattotalphenolcontentofacetoneextractofaerialpartofG.verumwas101.58±7.96mg GAE/g extract. This value is lower when compared to aqueous extract, whosecontentwas127.57±11.25mgGAE/gextract.Totalflavonoidcontentwas30.51±3.37and 61.24 ± 6.11 mg of quercetin/g of acetone and aqueous extracts, respectively.Furthermore examined extracts reduced DPPH with an IC50 11.65 ± 1.16 μg/ml ofacetoneand9.63±0.32μg/mlofaqueousextract.

ThesefindingdemonstratedthatbothextractsofG.verumrespresentsavaluablesource of phenols and flavonoids and indicated the rational basis for the traditionalusesofthisplantspeciesinalleviationofoxidativestress.Thisstudymaybeastartingpointforfurtherresearcheswhichwouldfullyexaminetheirchemicalcompositionaswellaspharmacologicaleffects.

688

Arh.farm 2018;68: 688 LB-P3

PHYTOCHEMICALSTUDYOFTHEGREEKENDEMICSPECIES

INULASUBFLOCCOSARECH.F.(ASTERACEAE)

DafniIoannaDiakaki,AikateriniKoutsaviti,EfstathiaIoannou,OlgaTzakou

SectionofPharmacognosyandChemistryofNaturalProducts,DepartmentofPharmacy,SchoolofHealthSciences,NationalandKapodistrianUniversityof

Athens(Greece)

Thegenus InulaL. (Asteraceae)comprisesc.90species,about19ofwhichare

native to Europe. They are perennial herbs, rarely shrubs, sometimes with anunpleasantsmell.SeveralInulaspeciesareusedastraditionalherbalmedicinestotreata broad spectrum of disorders, mainly respiratory, digestive, inflammatory,dermatological, aswell asmicrobial infections. Sesquiterpenes,mostly sesquiterpenelactonesincludingeudesmane,guaiane,pseudoguaianeandgermacranederivativesarecharacteristiccomponentsofInulaspecies,manyofwhichhaveexhibitedawiderangeof biological activities, particularly anti‐tumor and anti‐inflammatory. The chemicalprofile of Inula subfloccosaRech. f., an endemic plant of Greece that is restricted tocipolinandmarbles,hasnotbeenpreviouslystudied.

AerialpartsofI.subfloccosawerecollectedduringthefloweringperiod.Theair‐dried plant material was exhaustively extracted with CH2Cl2/MeOH at roomtemperature and the resulting crude extract was fractionated with a series ofchromatographicseparations.

So far, 9 sesquiterpene lactones have been isolated from the aerial parts of I.subfloccosa.Detailedanalysesof1Dand2DNMRandMSdataledtotheidentificationoffoureudesmanolides,fourguaianolides,andonepseudoguaianolide.

Amongtheisolatedcompounds,thebioactivemetabolites4α,5α‐epoxy‐10α,14H‐1‐epi‐ inuviscolide and isoalantolactone were the major constituents, whereas thepseudoguainolidederivativeisanewnaturalproduct.

689

Arh.farm 2018;68: 689 LB-P4 VOLATILECONSTITUENTSOFDIFFERENTPLANTPARTSOF

JUNIPERUSOXYCEDRUSSSP.MACROCARPA(SIBTH.&SM.)BALL.FROMGREECE

NikolaosArmenis,MariaCouladis

SectionofPharmacognosyandChemistryofNaturalProducts,DepartmentofPharmacy,SchoolofHealthSciences,NationalandKapodistrianUniversityof

Athens(Greece)

The family Cupressaceae is represented in Europe by five genera namely,

Cupressus, Chamaecyparis, Thuja, Tetraclinis and Juniperus. Their representatives aremonoecious or dioecious resiniferous trees or shrubs. Juniperus berries havetraditionallybeenusedfordyspepsia,acuteandchroniccystitis,arteriosclerosis,goutand inflammations, while essential oils of Juniperus species are used as fragranceingredientsincosmetics.Theaimofthepresentstudywastheanalysisofessentialoilsobtained from different plant parts of Juniperus oxycedrus ssp.Macrocarpa (Sibth. &Sm.)Ball.

Aerial parts of J. oxycedrus ssp. macrocarpa were collected from the Greekislands Chryssi andElafonissos. After hydrodistillation of the berry‐like fruits, leavesandbranches,theessentialoilswereobtained,respectively.Thechemicalcompositionofeachessentialoilwasdeterminedbygaschromatography(GC‐FID,GC‐MS).

Atotalof87componentswereidentifiedintheessentialoilofberry‐likefruits,representing 91.7% of the total oil composition. Themajor compoundwasα‐pinene(30.5%),followedbyα‐cedrol(10.4%),myrcene(10.1%)andgermacreneD(9.7%).Intheessentialoilof leaves113constituentswere identified,representing97.3%of thetotaloilcomposition.Themainconstituentwasα‐pinene(28.2%),followedbymanooloxide (12.5%) and α‐cedrol (9.8%). Finally, 107 compounds were identified in theessentialoilofthebranches,representing92.5%ofthetotaloilcomposition.Themajorcompound was manool oxide (20.5%), followed by α‐cedrol (14.4%) and α‐pinene(11.2%).Comparisonofourresults to the literaturedatashowedmainlyquantitativedifferences.

690

Arh.farm 2018;68: 690-691 LB-P5

ANATOMSKAGRAĐAIHEMIJSKAANALIZAETARSKOGULJA

PIMPINELLASAXIFRAGAL.(APIACEAE)

MilicaDrobac1,BojanZlatković2,NadaKovačević1,VioletaSlavkovska3

1Katedrazafarmakognoziju,UniverzitetuBeogradu‐Farmaceutskifakultet,2Departmanzabiologijuiekologiju,UniverzitetuNišu‐Prirodno‐matematičkifakultet,3Katedrazabotaniku,UniverzitetuBeogradu‐Farmaceutskifakultet

(Srbija)

Koren vrste Pimpinella saxifraga L. se tradicionalno koristi kao ekspektorans,

bronhosekretolitikiantiflogistik.Ciljovogradajeispitivanjeanatomskegrađe,sadržajai sastava etarskog ulja vegetativnih organa i ploda P. saxifraga. Biljni materijal jesakupljan u jugoistočnoj Srbiji: Bojanine vode (Suva planina) i Ostrovica (Sićevačkaklisura).Anatomskaanalizaizvršenajenatrajnimpreparatimadobijenimstandardnommetodom pripreme za posmatranje pod svetlosnim mikroskopom. Etarska ulja suizolovana iz herbe u cvetu, ploda, korena u fazi cvetanja i plodonošenja, destilacijomvodenomparomianaliziranaGC‐FIDiGC‐MSmetodama.

Rezultatianatomskeanalizesupokazalida jekorensekundarnegrađeastabloprimarnesazatvorenimkolateralnimsprovodnimsnopićima.Listovisudorziventralni,hipoamfistomatski, lisna drška je sa lučno raspređenim sprovodnim snopićima, aplodovi (merikarpi) su polukružnog oblika sa slabo izraženim rebrima, bez trihoma.Retke, nežlezdane, višećelijske, uniserijatne trihome sa oštrim vrhom su uočene nastablu,listovimailisnojdršci.Sekretornikanalisubrojniusvimorganima.

Najviši sadržaj etarskog ulja određen je u plodu (1,48‐1,52%), dok su herba(0,13‐0,21%)ikoren(0,50‐0,53%)sadržalimanjukoličinuulja(v/m).Etarskauljaistihorganasarazličitihlokalitetasusličnogkvalitativnogsastavasamanjimkvantitativnimrazlikama.Uuljimaherbeiplodadominantnajedinjenjasuβ‐bisabolen(28,8‐76,0%)iepoksi‐pseudoizoeugenil‐2‐metilbutirat (7,7‐21,8 %). Najzastupljenija jedinjenja uuljima korena, u obe faze, su azuleni (31,4‐35,7%) i pregeijeren (13,7‐18,8%). Uispitivanim etarskim uljima različitih organa P. saxifraga utvrđeno je prisustvotrinorseskviterpena (azulena i geijerena) i fenilpropanoidnih jedinjenjapseudoizoeugenil tipa,koja sukarakterističnihemijskimarkeri zaetarskauljabiljakaovogroda.

Istraživanje je podržalo Ministarstvo prosvete, nauke i tehnološkog razvoja

(ProjekatON173021).

691

ANATOMYANDCHEMICALANALYSISOFESSENTIALOILOF

PIMPINELLASAXIFRAGAL.(APIACEAE)

MilicaDrobac1,BojanZlatković2,NadaKovačević1,VioletaSlavkovska3

1DepartmentofPharmacognosy,UniversityofBelgrade‐FacultyofPharmacy,2DepartmentofBiologyandEcology,UniversityofNiš‐FacultyofSciencesand

Mathematics,3DepartmentofBotany,UniversityofBelgrade‐FacultyofPharmacy(Serbia)

Root of Pimpinella saxifraga L. is traditionally used as expectorant,

bronchosecretolyticandantiphlogistic.Theaimwasinvestigationofanatomy,contentandcompositionoftheessentialoilsofvegetativeorgansandfruits.Plantmaterialwascollected in South‐Eastern Serbia: Bojanine vode (Mt. Suva planina) and Ostrovica(Sićevogorge).Theanatomicalanalysiswasconductedonpermanentslidesobtainedby standard method of preparation. The essential oils obtained by hydrodistillationfromfloweringaerialparts,fruitsandroots(fromfloweringandfruitingperiod)wereanalysedbyGC‐FIDandGC‐MS.

The anatomical analysis revealed a secondary structure of root and primarystructureofstemwithclosedcollateralvascularbundles.The leavesaredorsiventral,hypoamphistomaticandpetioleiswitharchedvascularbundles.Thefruit(mericarp)issemi‐circular and lightly ribbed in cross section without trichomes. Rare, non‐glandular,multicellular, uniseriate, cuneate trihomeswere noted on the stem, leavesandpetiole.Secretorychannelsarenumerousinallorgans.

Thehighest contentof essentialoilwasdetermined in the fruits (1.48‐1.52%),whileaerialparts(0.13‐0.21%)androots(0.50‐0.53%)containedaloweramountofoil(v/w).Theoilsfromthesameorgansoriginatedfromdifferentlocalitiesweresimilarinqualitative composition with some quantitative differences. The most dominantconstituentsintheoilsfromaerialpartsandfruitswereβ‐bisabolene(28.8‐76.0%)andepoxy‐pseudoisoeugenyl‐2‐methylbutyrate(7.7‐21.8%),whereasthoseoftheoilsfromroots, in both stages, were azulenes (31.4‐35,7%) and pregeijerene (13.7‐18.8%).Trinorsesquiterpenes (azulenes and geijerenes) and phenylpropanoid compounds ofthe pseudoisoegenyl type,which are characteristic chemicalmarkers of the essentialoilsoftheplantsofthisgenus,werepresentintheallanalyzedoils.

ThestudywassupportedbytheMinistryofEducation,ScienceandTechnological

Development(ProjectON173021).

692

Arh.farm 2018;68: 692-693 LB-P6

ANTIMIKROBNAAKTIVNOSTSOKAPLODAARONIJE

(ARONIAMELANOCARPA(MICHX.)ELLIOTT)

MilicaMilutinović1,KatarinaŠavikin2,TatjanaMihajilovKrstev3,MilicaKostić1,BojanaMiladinović1,SuzanaBranković4,DušankaKitić1

1KatedrazaFarmaciju,UniverzitetuNišu‐Medicinskifakultet,2Institutzaproučavanjelekovitogbilja„DrJosifPančić”,3DepartmanzaBiologijui

Ekologiju,UniverzitetuNišu‐Pririodnomatematičkifakultet,4KatedrazaFiziologiju,UniverzitetuNišu‐Medicinskifakultet(Srbija)

Neosporna je činjenica da se otpornost mikroorganizama na poznate

antimikrobne lekove povećava. Stoga su različiti biljni proizvodi i njihovi aktivnisastojci u fokusu istraživanja kao potencijalni prirodni antimikrobni agensi. Plodovibiljne vrste Aronia melanocarpa (Michx.) Elliott (aronija) predstavljaju bogat izvorfarmakološki aktivnih jedinjenja kao što su polifenoli, što se povezuje saantimikrobnomaktivnošćukoju ispoljavaju. Cilj radabio jeutvrđivanje antimikrobneaktivnostisokaaronije.

Plodovi aronije sakupljeni su sa plantaža na planini Suvobor u Srbiji. Svežebobicesuzdrobljeneiisceđeneidobijenisokjefiltriran.Antimikrobnaaktivnostsokaaronijeprocenjenajekorišćenjemlaboratorijskihkontrolnihsojevamikroorganizama.Određivanje antimikrobne i antifungalne aktivnosti (minimalna inhibitornakoncentracija [MIC] i minimalna baktericidna/fungicidna koncentracija [MBC/MFC])dobijenajemikrodilucionommetodompremaNCCLS(2003).Minimalnakoncentracijaukojojnijebilovidljivog rastadefinisana jekaominimalna inhibitornakoncentracija(MIC).Minimalnabaktericidna/fungicidnakoncentracija(MBC/MFC)jedefinisanakaonajnižakoncentracijauzorkakoja jeubila99,9%ćelijamikroorganizama.Kontrolesuuključivale: medijum sa rastvaračem (negativnom kontrolom) i medijum sahloramfenikolom,streptomicinoministatinom(pozitivnakontrola).

Rezultati pokazuju da je sok aronije ispoljio antibakterijsku aktivnost protivGram‐pozitivnih bakterija Listeria monocytogenes, Bacillus cereus i Staphylococcusaureus,kaoiGram‐negativnihbakterijaSalmonellaenteritidisiEscherichiacoli.SoknijeimaouticajanaPseudomonasaeruginosa i sojevegljiva(Candidaalbicans iAspergillusniger).MinimalneinhibitorneimikrobicidnekoncentracijevarirajuodMIC/MBC=12,5do100mg/mL.PodaciizliteraturepotvrđujuantibakterijskuaktivnostplodovaaronijeprotivEscherichiacoli,Bacilluscereus,StaphylococcusaureusiPseudomonasaeruginosa.Sok aronije može biti koristan u prevenciji i lečenju bolesti izazvanih bakterijskiminfekcijama.

693

ANTIMICROBIALACTIVITYOFCHOKEBERRYJUICE

(ARONIAMELANOCARPA(MICHX.)ELLIOTT)

MilicaMilutinović1,KatarinaŠavikin2,TatjanaMihajilovKrstev3,MilicaKostić1,BojanaMiladinović1,SuzanaBranković4,DušankaKitić1

1DepartmentofPharmacy,UniversityofNiš‐FacultyofMedicine,2Institutefor

MedicinalPlantsResearch„DrJosifPancic”,3DepartmentofBiologyandEcology,UniversityofNiš‐FacultyofScienceandMathematics,4Departmentof

Physiology,UniversityofNiš‐FacultyofMedicine(Serbia)

It is an indisputable fact that themicrobial resistance to known antimicrobialdrugsincreases.Therefore,thevariousplantproductsandtheiractiveingredientsarein the focus of the research as a possible natural antimicrobial agent. The berries ofAronia melanocarpa (Michx.) Elliott (chokeberry) are the rich source ofpharmacological active compounds such as polyphenols and that may indicate theirantimicrobialactivity.Theaimoftheresearchwastoevaluatetheantimicrobialactivityofthechokeberryjuice.

Fruits of black chokeberrywere collected froma plantation field onmountainSuvobor,Serbia.Freshberrieswerecrushedandsqueezedand the juicewas filtered.The antimicrobial activity of black chokeberry juice was assessed using laboratorycontrol microorganism strains. The determination of antimicrobial and antifungalactivity (the minimum inhibitory concentration [MIC] and minimumbactericidal/fungicidal concentration [MBC/MFC]) was carried out by microwelldilution method according to the NCCLS (2003). The minimal concentration wheretherewasnovisiblegrowthwasdefinedastheminimalinhibitoryconcentration(MIC).The minimal bactericidal/fungicidal concentration (MBC/MFC) was defined as thelowestconcentrationof thesamplethathadkilled99.9%ofmicroorganismcells.Thecontrolsincluded:mediumwithsolvent(negative)andmediumwithchloramphenicol,streptomycinandnystatin(positive).

Results showed that chokeberry juice exhibited antibacterial activity againstGram‐positive bacteria Listeria monocytogenes, Bacillus cereus and Staphylococcusaureus, aswell as Gram–negative bacteria Salmonella enteritidis andEscherichia coli.The juice did not have influence on Pseudomonas aeruginosa and the fungal strains(Candida albicans and Aspergillus niger). The minimum inhibitory and microbicidalconcentrations ranged fromMIC/MBC=12.5 to 100mg/mL. Literaturedata showedantibacterial activity of chokeberries against Escherichia coli, Bacillus cereus,StaphylococcusaureusandPseudomonasaeruginosa.Chokeberry juicecouldbeusefulinthepreventionandtreatmentofthediseaseinducedbybacterialinfections.

694

Arh.farm 2018;68: 694-695 LB-P7

FITOHEMIJSKAANALIZAIODREĐIVANJESADRŽAJAFLAVONOIDAU

ALLIUMURSINUML.,ALLIACEAE

HarisNikšić,LejlaKadić,KemalDurić,IrmaSijamić,NadirJordamović

UniverzitetuSarajevu‐Farmaceutskifakultet(BosnaiHercegovina)

Kako bi se odredio sadržaj flavonida i antioksidativna svojstva etanolnih i

vodenih ekstrakata suhog i svježeg lista sa dva različita staništa srijemuša Alliumursinum L. u Bosni i Hercegovini, biljni uzorci su sakupljeni na dvije lokacije: Rogoj(1212mnadmorskevisine)iNeretvica(850mnadmorskevisine).

Esktrakti su pripremljeni od suhog i svježeg lista koristeći ekstrakciju napovišenoj temperaturi i ultrazvučnu ekstrakciju. Voda i 80% etanol su korišteni kaootapala.KorištenesuAlCl3iDPPHmetodazaodređivanjeantioksidativnogpotencijala.Nađeno je da sadržaj flavonoida ovisi o vrsti ekstrakcije i korištenom otapalu. Zadetekciju flavonoida korištena je TLC analiza. TLC analiza je potvrdila prisustvokvercetina,rutina,hiperozida,izokvercetinaikempferola.

TLCanalizapotvrdilajeznačajnijeprisustvokvercetinauetanolnimekstraktima.Sadržaj ukupnih flavonida kretao se u rasponu 11,31‐79,60 mg QE/g (suhog lista) i0,023‐32,054 mg QE/g (svježeg lista). Najveći sadržaj flavonoida pronađen je uetanolnom ekstraktu (suhog lista) 79,6 mgQE/g. Uzorak je prikupljen u Rogoju ipripremljen ultrazvučnom ekstrakcijom. Najmanji sadržaj flavonoida pronađen je uvodenom ekstraktu (svježeg lista) 0,023mgQE/g. Uzorak je prikupljen uNeretvici ipripremljenekstrakcijomnapovišenojtemperaturi.

Etanolni ekstrakti suhog lista ispoljili su najveći postotak inhibicije DPPH(30,80%). Uzorak je prikupljen u Rogoju. Etanolni ekstrakt svježeg lista ispoljio jenajmanjipostotakinhibicijeDPPHradikala(11,49%).UzorakjeprikupljenuRogoju.

695

PHYTOCHEMICALANALYSISANDFLAVONOIDCONTENTDETERMINATIONINALLIUMURSINUML.,ALLIACEAE

HarisNikšić,LejlaKadić,KemalDurić,IrmaSijamić,NadirJordamović

UniversityofSarajevo‐FacultyofPharmacy(BosniaandHerzegovina)

In order to estimate the amount of flavonoids and antioxidative properties of

ethanolicandwaterextracts fromdriedand fresh leavesof twodifferentecotypesofwildgarlic(AlliumursinumL.)inBosnaandHerzegovinaplantsampleswerecollectedontwoloctionsRogoj(altitude1212m)andNeretvica(altitude850m).

Extractswereprepared fromdry and fresh leavesusing extractionat elevatedtemperatureandultrasoundextraction.Waterand80%ethanolwereusedassolvents.AlCl3 and DPPH radical scavenging assay were used. It was found that the type ofextractionandsolventaffectthevalueofflavonoids.Fordeterminationofflavonoidsisused TLC. TLC analysis confirmed the presence of quercetin, rutin, hyperoside,isoquercetin,kaempherol.

The spot of quercetin was more intense in etanolic extracts. Total flavonoidcontentwas inrange11.31‐79.60mgQE/g(dried leaves)and0.023‐32.054mgQE/g(freshleaves).Thebiggestflavonoidcontentwasfoundinethanolicextract(dryleaves)79.6mgQE/g.Samplewascollected inRogojandpreparedbyultrasoundextraction.The smallest flavonoid content was found in water extract (fresh leaves) 0.023 mgQE/g. Sample was collected in Neretvica and prepared by extraction at elevatedtemperature.Nomatterwhichmethodofextractionwasused,higherflavonoidcontentwereinethanolicextracts.

Ethanolic extract of dry leaves had the biggest percent of inhibition of DPPHradical(30.80%).SamplewascollectedinRogoj.Ethanolicextractof freshleaveshadthe smallest percent inhibiton of DPPH radical (11.49 %). Sample was collected inRogoj.

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Arh.farm 2018;68: 696-697 LB-P8

ISPITIVANJEKVALITETABILJNIHDROGAKOJEULAZEUSASTAV

MONOKOMPONENTNIHČAJEVA

KemalDurić,MinelaGuta‐Mujezinović,HarisNikšić,IrmaSijamić,SamijaMuratović


Shodno propisima i metodama datim u važećoj Evropskoj farmakopeji u

poglavljuMetodefarmakognozijskeanalize,kaoipojedinimmonografijamaispitivanihdroga,predmetovestudijebio je ispitatikvalitetčetiribiljnedrogakojesekoristezaizradu raznih čajnih smjesa, a mogu biti samostalne, kao monokomponentni čajevi,shodno propisima i metodama datim u Evropskoj farmakopeji u poglavljuFarmakognostičkaispitivanja.

Ispitivanjejeprovedenonačetribiljnedroge,nabavljeneizdvarazličitaizvora,tenabiljnommaterijalukojesemoženaćiuslobodnojprodajipodidentičnimimenomkao ranije spomenute biljne droge. Ispitivane biljne droge bile su: Absinthii herba(Artemisia absinthium L.), Melissae folium (Melissa officinalis L.), Menthae piperitaefolium (MenthapiperitaL.) iThymiherba (Thymus vulgarisL.). Općemetode koje sukorištenezaanalizubilesu:makroskopskaanaliza,mikroskopskaanaliza,određivanjesadržaja stranih supstanci, određivanje sadržaja vode i određivanje sadržaja pepela.Specijalnemetodekojesukorištenezaanalizubilesu:određivanjesadržajaeteričnihulja,određivanjegorčinetetankoslojnahromatografija.

Ispitivanebiljnedroge,nabavljeneuzvaničnojprodaji,pokazalesuse izuzetnokvalitetnim, s obzirom da su vrijednosti dobijene ispitivanjem parametara kvalitetezadovoljavale farmakopejom definirane kriterijume. Biljni materijal nabavljen uslobodnoj, nezvaničnoj prodaji pokazao je najviše propusta u pogledu ispitivanjakvalitete.

Dobijenirezultatiukazujunaopravdanostivažnostfarmakognozijskeanalizeupogledukvalitetebiljnihmaterijalakojisekoristeuslužbenojfarmacijiimedicini.

697

QUALITYTESTINGOFHERBALDRUGSINCLUDEDINTHE

COMPOSITIONOFMONOCOMPONENTTEAS

KemalDurić,MinelaGuta‐Mujezinović,HarisNikšić,IrmaSijamić,Samija Muratović


The subject of this studywas in accordancewith the regulations andmethods

givenbythecurrentlyvalidEuropeanPharmacopoeiainthechapterPharmacognosticexaminations,aswellascertainmonographsofinvestigateddrugs:Examinethequalityofthefourtypesofdrugs,whichareincompositionofvariousteamixturesandmaybeself‐contained as mono‐constituent teas, according to the regulations and methodsgivenbyEuropeanPharmacopoeiainthechapterPharmacognosy.

Thestudywasconductedonfourherbaldrugsfromtwodifferentsourcesandonherbalmaterialwhichcanbeobtainedonafreemarketbythenameequivalenttothepreviously mentioned herbal drugs. Tested drugs were: Absinthii herba (ArtemisiaabsinthiumL.),Melissaefolium(MelissaofficinalisL.),Menthaepiperitaefolium(Menthapiperita L.) andThymiherba (Thymusvulgaris L.).Generalmethodsused for analysiswere:macroscopicalanalysis,microscopicalanalysis,determinationofforeignmatters,determinationofwatercontentanddeterminationofash.Specificmethodsusedwere:determination of essential oils content, determination of bitterness and thin layerchromatography.

Herbal drugs from official sources shown a very good quality rate, since theobtained values by examination of quality parameters met pharmacopoeias definedcriteria. Herbal material from free sale and non‐official source, shown the lowestqualityandwasnotinaccordancewithamostofPharmacopoeialdemands.Obtainedresultsindicatesthevalidityandimportanceofpharmacognosticanalysisregardingthequalityofherbalmaterialsanddrugsusedinofficialpharmacyandmedicine.

698

Arh.farm 2018;68: 698-699 LB-P9

FITOHEMIJSKOISPITIVANJETRITERPENSKIHSAPONINASA

POTENCIJALNIMFARMAKOLOŠKIMDJELOVANJEMUVRSTAMAPORODICELAMIACEAE

NadirJordamović,HarisNikšić,IrmaSijamić,KemalDurić


Triterpenskiderivatiprisutniubiljkamasučestospojevinosiocifarmakološkog

djelovanja. Ova grupa obuhvata različite spojeve od kojih su najvažniji pentacikličnidrivatioleana,ursana, lupanaitetracikličnidamarani.Ispitivanebiljnevrstesudobropoznate ikoristese jersadržeeteričnoulje, fenolnespojeve idrugemetabolitealinezbogprisustvabetulina,betulinske,oleanolne,ursolnekiselineilupeola.Prisustvoovihvažnih spojeva bi moglo odrediti neke plejotropne farmakološke učinke ovih biljnihdroga.

U ovom radu je provedena kvalitativna ii kvantitativna analiza triterpenskihspojevaubiljnimvrstamaporodiceLamiaceae.Ciljstudijebiopronaćinajboljeuslovekvantitativne ekstrakcije ispitivanih triterpenskih supstanci što je postignutosukcesivnomekstrakcijomotapalimarazličitepolarnosti. Identifikacija ikvantifikacijaciljnih spojeva u različitim frakcijama je postignuta primjenom metoda TLC i HPLC.Standardi triterpenskih supstanci (betulin, betulinska, oleanolna, ursolna kiselina ilupeol) i pojedine frakcije su nanošne koristeći CAMAG Linomat 5 aparat. Sistem zarazvijanje (benzen: etilacetat: mravlja kisleina – 36:12:5) i uslovi HPLC metode(ConstaMetric®3000sistem,HypersilODS(AgilentTechnologies)kolona,4.6x250mm,5μm, mobilna faza acetonitril/voda (700/300) acidifikovana sa orto‐fosfornomkiselinom)susepokazalioptimalnizaseparaciju triterpenskihsupstanci,koji suvrlosličnehemijskestrukture.

Dobijeni rezultati potvrđujuprisustvo ovih važnih supstanci u biljnimvrstamakoje pripadaju porodici Lamiaceae, sakupljene na području Bosne i Hercegovine.Najvećisadržajbetulina(4,19µg/ml)ibetulinskekiseline(51,58µg/ml)pronađenisuuheksanskojfrakcijilistaruzmarina.Hloroformskiekstraktcvijetalavandejepokazaonajvećukoncentracijuursolnekiseline(0,14mg/ml).Oleanolnakiselinajebilaprisutnasamouheksanskomekstraktulistaruzmarina(1,01µg/ml).Listruzmarinapredstavljapotencijalnufarmakognostičkudrogusastanovištaprisustvatriterpenskihsaponina.

699

PHYTOCHEMICALINVESTIGATIONOFTRITERPENESAPONINS

WITHPOTENTIALPHARMACOLOGICALACTIVITYINPLANTSPECIESBELONGINGTOLAMIACEAEFAMILY

NadirJordamović,HarisNikšić,IrmaSijamić,KemalDurić


Triterpene derivatives in plants are compounds that are frequently bearersof

pharmacologicalactivities.Thisgroupcomprisedifferentcompounds,amongwhichthemost important are pentacyclic derivatives of olean, ursane, lupane and tetracyclicdamaranes. Investigated plant species are well known and used for their content inessentialoil, phenol compounds, andothermetabolomes,butnot for thepresenceofbetulinic, oleanolic, lupeol, and ursolic acid. Identification of this compounds coulddeterminatesomepleiotropicpharmacologicalactivityofthisherbaldrugs.Qualitativeandquantitativeanalysisoftriterpenes,presentinplantspeciesbelongingtoLamiacaefamily was performed in this study. The aim of this study was to find out the bestconditions forquantitativeextractionof investigatedtriterpenesubstanceswhichhasbeenachievedbysuccessiveextractionusingdifferentpolarsolvents.

Identificationandquantificationoftargetcompoundsindifferent fractionswasobtained using TLC and HPLC method. Triterpene standards (betulin, betulinic,oleanolic, lupeol, andursolicacid)anddifferent fractionswereappliedonTLCplatesusing»CAMAGLinomat5«device.TheSystemsused fordeveloping inTLC(benzene:ethylacetate:formicacid(36:12:5)andHPLCmethod(ConstaMetric®3000systemforsolvent release, Hypersil ODS (Agilent Technologies) column, 4.6 x 250 mm, 5μm,mobile phase acetonitril/aqua (700/300) acidifiedwith ortho‐phosphoric acid)wereprovedtobeoptimalforseparationoftestedtriterpenesubstances,otherwisehavingverysimilarchemicalstructure.

The results confirm the presence of this important compounds in the plantspecies belonging to Lamiaceae family collected from the Bosnia and Herzegovinaregion.Thehighestamountofbetulin (4.19µg/ml)andbetulinicacid (51.58µg/ml)werefoundinhexaneextractofRosmarinifolium.ChloroformextractofLavandulaeflosshowthehighestamountofursolicacid(0.14mg/ml).OleanolicacidwasfoundonlyinthehexaneextractofRosmarinifolium(1.01µg/ml).DuetothepresenceoftriterpenesubstancesRosmarinifoliumcouldrepresentpotentialpharmacognosticdrug.

700

Arh.farm 2018;68: 700-701 LB-P10

ISPITIVANJEMAKROSKOPSKIH,MIKROSKOPSKIHOSOBINAIETARSKOGULJALISTAMATIČNJAKA,MELISSAOFFICINALISL.

MirjanaMarčetić1,NikolaBošković1,DaniloStojanović2,NadaKovačević1

1Katedrazafarmakognoziju,UniverzitetuBeogradu‐Farmaceutskifakultet,2Katedrazabotaniku,UniverzitetuBeogradu‐Farmaceutskifakultet(Srbija)

Tradicionalno list matičnjaka,Melissae folium, koristi se za ublažavanje blagih

simptomamentalnogstresaiolakšavanjeuspavljivanja,kaoiusimptomatskojterapijiblagih gastrointestinalnih poremećaja. Etarsko ulje matičnjaka je veoma cenjeno uprehrambenoj i kozmetičkoj industriji. Cilj rada je bio ispitivanje makroskopskih imikroskopskih osobina uzoraka lista samoniklog i gajenog matičnjaka sa različitihlokaliteta i komercijalno dostupnih čajeva, kao i ispitivanja sastava njihovih etarskihulja.

Za analizu etarskih ulja (16 uzoraka), dobijenih destilacijom vodenom parom,primenjena je gasna hromatografija (GC‐FID) i gasna hromatografija sa masenomspektrometrijom(GC‐MS).Klasteranalizomispitivanesurazlikeizmeđuetarskihulja.

Makroskopske i mikroskopske odlike svih uzoraka su odgovarale monografijiMelissae folium u Ph. Eur. 7.0, ali je u 2 od 5 komercijalnih uzoraka bio veći sadržajstranih materija. U svim ispitivanim etarskim uljima glavne komponenate supredstavljali: geranial (15,58‐53,33%), neral (10,73‐36,64%), kariofilen oksid (0,76‐35,57%),(E)‐kariofilen(tragovi‐16,72%)icitronelal(0,00‐12,64%).Sadržajgeranialai nerala u ispitivanim uzorcima je bio u opsegu vrednosti dobijenih u prethodnimispitivanjima.

Klaster analizom uzorci etarskog ulja su podeljeni na dva klastera. U prvomklasteru subila etarskaulja sa visokim sadržajemgeraniala i nerala (31,87‐53,33% i19,81‐36,64%):etarskaulja lista iherbesamoniklog igajenogmatičnjakaurazličitimfazama razvoja i sa različitih lokaliteta i dva komercijalna uzorka.U drugi klaster susvrstana etarska ulja sa nižim sadržajem geraniala i nerala (15,58‐26,67% i 10,73‐15,48%) ivećimudelomkariofilen‐oksida(17,79‐35,57%):trikomercijalnauzorka ietarskouljeherbe.Listsamoniklogmatičnjakaurazličitimfazamarazvoja,sadrživisokprocenatgeranialaineralauetarskomulju,nerazlikujeseznačajnoodgajenihuzorakaimožepredstavljatiizvorkvalitetnedroge.


(ProjekatON173021).

701

INVESTIGATIONOFMACROSCOPICANDMICROSCOPIC

CHARACTERISTICSANDESSENTIALOILOFTHELEMONBALMLEAF,MELISSAOFFICINALISL.

MirjanaMarčetić1,NikolaBošković1,DaniloStojanović2,NadaKovačević1

1DepartmentofPharmacognosy,UniversityofBelgrade‐FacultyofPharmacy,2DepartmentofBotany,UniversityofBelgrade‐FacultyofPharmacy(Serbia)

Traditionalherbalremediesbasedonlemonbalmleaf,Melissaefoliumareused

for relief of mild symptoms of mental stress, to aid sleep and for symptomatictreatmentofmildgastrointestinalcomplaints.Theessentialoilof lemonbalm isveryappreciated in foodand cosmetic industry.The aimwas to examine themacroscopicandmicroscopiccharacteristicsoflemonbalmleavesofcultivatedandwildplantsfromdifferent localities, commercially available tea samples and to determinate thecompositionoftheiressentialoils.Theessentialoilsobtainedbyhydrodistillation(16samples), were analysed by gas chromatography (GC‐FID) and gas chromatographywith mass spectrometry (GC‐MS). The differences between essential oils wereexaminedbyclusteranalysis.

Macroscopic and microscopic characteristics corresponded to the monographMelissae folium in Ph. Eur. 7.0, while 2 of 5 commercial samples contained highercontentofforeignmatter.

Themaincomponentsininvestigatedessentialoilswere:geranial(15.58‐53.33%), neral (10.73‐36.64 %), caryophyllene oxide (0.76‐35.57 %), (E)‐caryophyllene(traces‐16.72%)andcitronellal(0.00‐12.64%).Thecontentofgeranialandneralwasintherangeofvaluesobtainedinpreviousinvestigations.

Clusteranalysisdividedsamplesofessentialoilsintotwoclusters.Essentialoilswithhighgeranialandneralcontent(31.87‐53.33%and19.81‐36.64%)were in thefirst cluster: essential oils of leaf and aerial parts ofwild and cultivated lemon balmfrom different development phases and localities and two commercial samples. Theessentialoilswithlowergeranialandneralcontent(15.58‐26.67%and10.73‐15.48%)andhighercaryophylleneoxide(17.79‐35.57%)wereseparatedtothesecondcluster:threecommercialsamplesandessentialoilofaerialparts.Thewildlemonbalmleafindifferent phases of development contains essential oilwith high amounts of geranialandneral.Itisnotsignificantlydifferentfromcultivatedsamplesandcouldrepresentasourceofqualityherbalsubstance.



702

Arh.farm 2018;68: 702-703 LB-P11

MASNEKISELINE,STEROLIITRITERPENIMASNIHULJAPLODOVAOSAMTAKSONARODAHERACLEUML.IZJUGOISTOČNEEVROPE

LjubošUšjak1,IvanaSofrenić2,VeleTešević2,MilicaDrobac1,

MarjanNiketić3,SilvanaPetrović1

1Katedrazafarmakognoziju,UniverzitetuBeogradu‐Farmaceutskifakultet,2UniverzitetuBeogradu‐Hemijskifakultet,3Prirodnjačkimuzej(Srbija)

TaksonirodaHeracleumL.surasprostranjenipredstavnicifamilijeApiaceae,koji

bi mogli biti od značaja u raznim granama industrije. Predmet ovog ispitivanja sumasnekiseline,steroliitriterpenimasnihuljaplodovaH.sphondyliumL.,H.sibiricumL.,H.montanumSchleich.exGaudin,H.ternatumVelen.,H.pyrenaicumsubsp.pollinianum(Bertol.) F. Pedrotti & Pignatti, H. pyrenaicum subsp. orsinii (Guss.) F. Pedrotti &Pignatti,H.verticillatumPančić iH.orphanidisBoiss.,sakupljenihuSrbiji,Makedoniji,CrnojGoriiSloveniji.

Osušeni i samleveni plodovi ekstrahovani su dihlormetanom postupkombimaceracije na sobnoj temperaturi. Rastvarač je uklonjen pod sniženim pritiskom,ekstrakti filtrirani i dobijeni supernatanti podvrgnuti postupku saponifikacije.Saponifikovanefrakcije,bogatemasnimkiselinama,sumetilovane,anesaponifikovanefrakcije, koje prvenstveno sadrže sterole i triterpene, silanizovane, u cilju dobijanjaisparljivih derivata, koji su analizirani gasnom hromatografijom (GC‐FID i GC‐MS).Komponente su identifikovane pomoćukomercijalnih standarda i bibliotekamasenihspektara.

Šesnaestodukupnoosamnaestidentifikovanihmasnihkiselinasudetektovaneusvim saponifikovanim frakcijama, a dominantne su bile petroselinska (42,8‐56,5%),linolna (20,3‐33,3 %) i oleinska (12,3‐13,7 %) kiselina. U svakoj nesaponifikovanojfrakcijiidentifikovanisutriterpenα‐amirin(0,8‐6,0%)idevetistihsterola,odkojihjenajzastupljeniji bioβ‐sitosterol (44,9‐56,9%), a sledili su ga po količini stigmasterol(15,7‐25,0 %), Δ7‐stigmastenol (6,6‐12,5 %) i kampesterol (5,2‐8,1 %). Kaonajznačajnije može se istaći prisustvo petroselinske kiseline, potencijalno važnesirovinezafarmaceutsku,kozmetičku,prehrambenuihemijskuindustriju.

Uovomradu,masnekiseline,steroliitriterpenimasnihuljaplodovaH.ternatum,H.pyrenaicum subsp.pollinianum,H.verticillatum iH.orphanidis ispitivanisupoprviput,auslučajuH.sphondylium,H.sibiricum,H.montanumiH.pyrenaicumsubsp.orsinii,postojeći podaci o ovim sastojcima masnih ulja su značajno dopunjeni. Dobijenirezultati pružaju dobru osnovu za dalja istraživanja, u cilju primene ovih biljaka kaopotencijalnihnovihizvoraindustrijskiznačajnihmasnihulja.


(ProjektiON173021;ON172053).

703

FATTYACIDS,STEROLSANDTRITERPENESOFTHEFRUITFATTY

OILSOFEIGHTHERACLEUML.TAXAFROMSOUTHEASTERNEUROPE

LjubošUšjak1,IvanaSofrenić2,VeleTešević2,MilicaDrobac1,MarjanNiketić3,SilvanaPetrović1

1DepartmentofPharmacognosy,UniversityofBelgrade‐FacultyofPharmacy,

2UniversityofBelgrade‐FacultyofChemistry,3NaturalHistoryMuseum(Serbia)

HeracleumL. taxa arewidespreadmembers of Apiaceae family,with potential

application in different industries. Focus of this studywason fatty acids, sterols andtriterpenes of the fruit fatty oils ofH. sphondylium L.,H. sibiricum L.,H.montanumSchleich. exGaudin,H. ternatum Velen.,H.pyrenaicum subsp.pollinianum (Bertol.) F.Pedrotti & Pignatti, H. pyrenaicum subsp. orsinii (Guss.) F. Pedrotti & Pignatti, H.verticillatum Pančić and H. orphanidis Boiss., collected in Serbia, Macedonia,MontenegroandSlovenia.

Air‐driedandpowderedfruitswerebimaceratedwithdichloromethaneatroomtemperature. Solventwas evaporated under reduced pressure, extractswere filteredandobtainedoilyresiduesweresubjectedtosaponification.Saponifiablefractions,richin fatty acids, were further subjected to methylation, and unsaponifiable fractions,mostlycontainingsterolsandtriterpenes,tosilanization,toobtainvolatilederivativesthat were analyzed by GC‐FID and GC‐MS. Compounds were identified usingcommercialstandardsandmassspectralibraries.

Sixteen of the total of eighteen identified fatty acids were detected in allsaponifiablefractions,withpetroselinic(42.8‐56.5%),linoleic(20.3‐33.3%)andoleic(12.3‐13.7 %) acids being the most abundant. In every unsaponifiable fraction, atriterpeneα‐amyrin(0.8‐6.0%)andthesameninesterols,predominantlyβ‐sitosterol(44.9‐56.9%), followed by stigmasterol (15.7‐25.0%), Δ7‐stigmastenol (6.6‐12.5%)andcampesterol(5.2‐8.1%),wereidentified.Themostnotably,petroselinicacidcanbeutilizedinpharmaceutical,cosmetic,foodandchemicalindustries.

In this research, fatty acids, sterols and triterpenes of the fruit fatty oils ofH.ternatum,H. pyrenaicum subsp. pollinianum,H. verticillatum andH. orphanidis wereinvestigated for the first time, while in the case of H. sphondylium, H. sibiricum, H.montanum and H. pyrenaicum subsp. orsinii, the knowledge about their fatty oilsconstituents was enhanced. Obtained results provide a good basis for furtherinvestigations, aiming to establish theseplants aspotential sourcesof valuablenovelrawmaterials.


Development(ProjectsON173021;ON172053).

704

Arh.farm 2018;68: 704-705 LB-P12 ANTIINFLAMATORNIEFEKATEKSTRAKTAMUSKATNEŽALFIJE

(SALVIASCLAREAL.)

MilicaKostić1,DušankaKitić1,MilicaB.Petrović2,TatjanaJevtović‐Stoimenov3,BojanaMiladinović1,

MilicaMilutinović1,BojanZlatković4

1Katedrazafarmaciju,UniverzitetuNišu‐Medicinskifakultet,2UniverzitetuNišu‐Medicinskifakultet,3Katedrazabiohemiju,Medicinskifakultet,UniverzitetuNišu,4Departmanzabiologijuiekologiju,Univerzitetu

Nišu‐Prirodno‐matematickifakultet(Srbija)

SalviasclareaL.jearomatičnabiljnavrstačijaselekovitostzasnivanaodličnim

antimikrobnim, antiinflamatornim, antioksidativnim i spazmolitičkim efektima.Najčešćeseusuvomilisvežemoblikukoristikaostomahik,kaoiutretmanugingivitisa,stomatitisaiafti.CiljradajeutvrditiantiinflamatornuaktivnostekstraktaS.sclareanamodelueksperimenatalnoindukovanogperiodontitisapacova.

Inflamacija periodoncijuma indukovana je lipopolisaharidom (LPS) u prostoruizmeđuprvogidrugogdesnogmaksilarnogmolarapacova.Etanolniekstraktmuskatnežalfije, pripremljen ultrazvučnom ekstrakcijom herbe, primenjivan jeper os (200mg/kg) dva puta dnevno tokom deset dana. Grupu I i II činili su pacovi kojima jeubrizganfiziološkirastvor,atretiranivodomiliekstraktom,dokjegrupamaIII,IViVubrizganLPSrastvor,a tretiranisuvodom,ekstraktom iliekstraktompreventivnotridana pre indukovanja inflmacije, redom. Status inflamacije praćen je merenjemproinflamatornihcitokinaugingivalnomtkivuELISAtestovima:interleukina‐1beta(IL‐1beta), interleukina‐6 (IL‐6) i faktora tumorske nekroze‐alfa (TNF‐alfa). Ekstrakt jehemijskiokarakterisanHPLChromatografijom.

U grupi pacova koja je bila tretirana ekstraktom (IV i V) je značajno došlo doredukovanjanivoacitokinaupoređenjusanetretiranomgrupom(III):IL‐1beta(1,15i0,54vs.2,16pg/mg), IL‐6(5,58i5,05vs.6,93pg/mg)iTNF‐alfa(0,27i0,29vs.4,35pg/mg). Preventivna aplikacija ekstrakta nije bila od značaja. U grupama I i II nisuzabeleženi povećani nivoi citokina. U ekstraktu je ruzmarinska kiselina bilapredominantna(165,30ug/mg),aumanjojkoličinisubilezastupljeneikafenakiselina,luteolin, luteolin‐7‐O‐glukozid, apigenin i apigenin‐7‐O‐glukozid. Ekstrakt muskatnežalfijeispoljavaantiinflamatorniefekatnamodeluLPS‐indukovanogperiodontitisakodpacova, pa stoga može imati ulogu potencijalnog i značajnog terapijskog sredstva utretmanuperiodontalnihbolesti.


(III41018iIII46013).

705

THEANTI‐INFLAMATORYEFFECTOFTHECLARYSAGEEXTRACT

(SALVIASCLAREAL.)

MilicaKostić1,DušankaKitić1,MilicaB.Petrović2,TatjanaJevtović‐Stoimenov3,BojanaMiladinović1,

MilicaMilutinović1,BojanZlatković4

1DepartmentofPharmacy,UniversityofNiš‐FacultyofMedicine,2UniversityofNiš‐FacultyofMedicine,3DepartmentofBiochemistry,UniversityofNiš‐FacultyofMedicine,4DepartmentofBiologyandEcology,UniversityofNiš‐

FacultyofSciencesandMathematics(Serbia)

SalviasclareaL.isanaromaticplantspecieswhosehealingpropertiesarebased

onexcellentantimicrobial,anti‐inflammatory,antioxidantandspasmolyticeffects.Itismost often used dry or fresh as a stomachic, but also in gingivitis, stomatitis andaphthaetreatment.Theaimofthepaperistodeterminetheanti‐inflammatoryactivityofaS.sclareaextractontheexperimentallyinducedperiodontitisinrats.

Periodontitiswas inducedby lipopolysaccharide(LPS) in theareabetweenthefirstandthesecondrightmaxillaryratmolars.Anethanolextractofsage,preparedbyultrasonicextractionoftheherb,wasadministeredperos (200mg/kg)twicedailyfortendays.GroupIandIIwereinjectedwithsalineandtreatedwithwaterortheextract,whilegroupsIII,IVandVwereinjectedwithLPSandtreatedwithwater,theextractorthe extract threedays before inflammation induction, respectively. The inflammationstatus was monitored by proinflammatory cytokines measurements (ELISA) in thegingival tissue: interleukin‐1beta(IL‐1beta), interleukin‐6 (IL‐6) and tumor necrosisfactor‐alfa(TNF‐alfa).TheextractwaschemicallycharacterizedbyHPLC.

Asignificantreduction incytokine levelswasobserved intheratstreatedwiththeextract(IVandV),comparedtotheuntreatedgroup(III):IL‐1beta(1.15and0.54vs.2.16pg/mg), IL‐6 (5.58and5.05vs.6.93pg/mg)andTNF‐alfa (0.27and0.29vs.4.35pg/mg).Thepreventiveapplicationoftheextractwasnotsignificant.InthegroupIandIIcytokineslevelswerenotincreased.Rosmarinicacidwaspredominant(165.30ug/mg) intheextract,whilecaffeicacid, luteolin, luteolin‐7‐O‐glucoside,apigeninandapigenin‐7‐O‐glucoside were present in smaller amounts. The clary sage extractexhibitsananti‐inflammatoryeffectontheLPS‐inducedperiodontitis inrats,andcanthereforebeusedasapotential andsignificant therapeuticagent in the treatmentofperiodontaldiseases.


Development(III41018andIII46013)

706

Arh.farm 2018;68: 706-707 LB-P13

KONTROLAKVALITETAKOREISPRAŠENEKORECIMETANA

TRŽIŠTUSRBIJE

KatarinaJeremić,NedaGavarić,NebojšaKladar,NemanjaTodorović,NebojšaSalaj,MajaBekut,BiljanaBožin

Katedrazafarmaciju,UniverzitetuNovomSadu‐Medicinskifakultet(Srbija)

Invitroiinvivoistraživanjacimetaukazujunavisokmedicinskipotencijalovog

začina.Značajnadelovanjasuantimikrobno,antiinflamatorno,uticajnasniženjenivoaholesterola,analaziseikaosastojakdijetetskihsuplemenatanamenjenihzakoterapijui prevenciju šećernebolesti. Zvanični ispitani izvor ovedroge jeCinnamomumverum(zeylanicum)‐cejlonski cimet, dok se na tržištu najčešće nalazi Cinnamomum cassia‐kineskicimetiCinnamomumburmanii‐ indonežanskicimet.Ovevrsteserazlikujupokvalitetuibezbednostiupotrebe.CiljovogradabiojedaseispitakvalitetcimetakojijedostupannatržištuSrbijepremaPravilnikuokvalitetuzačina.

UovomraduzaizolacijuetarskoguljaprimenjenajemetodahidrodestilacijepoPh.Jug.V.Količinaetarskoguljaizrazisekaoprosečnavrednostprinosaiztrimerenjapomasisuvogbiljnogmaterijala(g/100g).UkupanpepeoipepeonerastvorljivuHClodređen je prema Ph. Jug. V. Izražen je u procentima. Rezultati su poređeni sapropisimavažećegPravilnikaokvalitetuzačinaRepublikeSrbije.

Odsedamispitivanihuzorakasamosprašenakoraorganskigajenogcejlonskogcimeta zadovoljava propis u pogledu sadržaja etarskog ulja, dok s druge strane koracejlonskog cimeta odstupa od propisanog minimalnog sadržaja cimeta čak 7,5 puta,najviše od svih ostalih uzoraka (kora i sprašena kora indonežanskog cimeta korakineskogcimeta).PrimetnajerazlikausadržajuuljakodIndonežanskogcimetauprahukoji ječuvanunajlonskojambalaži(0,36%) ionogkoji ječuvanupapirnojambalaži(0,15 %). Svih sedam uzoraka zadovoljava propis za maksimalan sadržaj ukupnogpepelaipepelanerstvornoguHCl.

Sobziromnaširokuupotrebucimeta,kakokaozačina,takoikaododatkaishranisciljempostizanjaodređenihzdravstvenihbenefita,njegovuvoziidentifikacijatrebalabidapodležestrožijimkriterijumimakakobiseizbegaopotencijalanrizikpozdravlje.

707

QUALITYCONTROLOFCINNAMON’SBARKANDPOWDEREDBARK

FROMTHESERBIANMARKET

KatarinaJeremić,NedaGavarić,NebojšaKladar,NemanjaTodorović,NebojšaSalaj,MajaBekut,BiljanaBožin

DepartmentofPharmacy,UniversityofNoviSad‐FacultyofMedicine(Serbia)

Invitroandinvivoresearchindicatethehighmedicalpotentialofthisspice.The

problemwiththeuseofthisplantisthattherearehundredsofspeciesandsubspeciesof cinnamon plant. The official source is Cinnamomum verum (zeylanicum) ‐ Ceyloncinnamon,whileCinnamomumcassia‐ChinesecinnamonandCinnamomumburmanii‐Indonesiancinnamon,arecommonlyfoundonthemarket.Itisimportanttoemphasizethattheydiffer inqualityandsafetyofuse.TheaimofthispaperwastoexaminethequalityofcinnamonthatisavailableontheSerbianmarketaccordingtotheRulesforthequalityofspices.

InthisstudyessentialoilwasisolatedusinghydrodestilationmethodaccordingtoPh.Yug.V,whosequantityisexpressedinpercentages.TotalashandashinsolubleinHClwasdeterminedaccordingtoPh.Yug.V.Itisexpressedinpercentages,too.

Only the powder of the organic Ceylon cinnamon meets the standard of theregulationforthecontentofessentialoil,whileontheotherside,barkoftheCeyloniancinnamonhasthebiggestdeviationfromtheprescribedminimumcontent,mostofallothersamples(IndonesianpowderandbarkandChinesebark).There isasignificantdifferenceintheoilcontentofIndonesiancinnamonpowderstoredinnylonpackaging(0.36 %) and the one stored in paper packaging (0.15 %). All seven samples havesatisfiedtheregulationforthemaximumcontentoftotalashandashinsolubleinHCl.

Cinnamoniswidelyusedasanadditiontonutritionforachievinghealthbenefits.Therefore,itsimportsandidentificationshouldbesubjecttostrictercriteriainordertoavoidpotentialhealthrisks.

708

Arh.farm 2018;68: 708-709 LB-P14

VAZORELAKSANTNAAKTIVNOSTTERPINOLENA

ReljaSuručić1,DraganaDrakul2,NadaKovačević3,

TatjanaKundaković‐Vasović3

1Katedrazafarmakognoziju,UniverzitetuBanjojLuci‐Medicinskifakultet,RepublikaSrpska,2UniverzitetuIstočnomSarajevu‐Medicinskifakultet,RepublikaSrpska,(BosnaiHercegovina),3Katedrazafarmakognoziju,

UniverzitetuBeogradu‐Farmaceutskifakultet(Srbija)

Terpeni predstavljaju veoma važnu grupu hemijskih jedinjenja kako zbogindustrijske upotrebe, tako i zbog svojih značajnih bioloških efekata koji se moguiskoristitiumedicini.Jednasuodnajbrojnijihgrupasekundarnihmetabolitabiljaka,anazivoveklase jedinjenjapotičeodterpentina(terpentinskogulja), tečnogproizvodadestilacijeoleorezinabora.Terpinolen(p‐menta‐2,4(8)‐dien)jebezbojnailibledožutoobojenatečnostaromatičnogmirisa.Ovajmonociklični,monoterpenskialkenješirokozastupljen u biljnim tkivima različitih četinara, paškanata, konoplje, čajevca, listukurkumeiperšuna,ačestjesastojaketarskoguljaCitrus,Mentha,JuniperusiMyristicavrsta. Jedna je od glavnih komponenata etarskog ulja nadzemnih delova endemičnevrste Seseli gracile Waldst. & Kit. (6,1‐57,5 %). Terpinolen poseduje potvrđenuantiradikalsku, antihiperalgezijsku i antiedematoznu aktivnost. S obziromda su nekastrukturno slična monoterpenska jedinjenja pokazala vazorelaksantnu aktivnost, ciljovogistraživanjajedaseistraživazorelaksantnipotencijalterpinolena.

U ovom istraživanju ispitivan je efekat standarda terpinolena (43905 Sigma‐Aldrich) na izolovanoj renalnoj arteriji pacova Wistar soja. Korišćeni su prstenastisegmentirenalnearterijedužine3‐5mmsaočuvanimendotelom.Arterijskisegmentisu postavljani u vodeno kupatilo u Krebs‐Ringerov rastvor, na 37C i aerisanikontinuiranim dovođenjem smješe 95%O2 i 5% CO2. Kontrakcija krvnih sudova jeizazvanaprimjenomfenilefrina(10‐6M).Integritetendotelajepotvrđenfarmakološki,primjenomacetilholina(10‐6M).Nastabilantonuskrvnogsudaizazvanfenilefrinomsudodavane rastuće koncentracije terpinolena, pripremljenog od standardarazblaživanjem u 5% karboksi‐metil celuloze (0,2% ‐ 33,3%, kumulativno). Svakasledeća koncentracija je dodavana tek nakon što se završi efekat prethodnoprimjenjene koncentracije, tj. otprilike nakon 15minuta od prethodne. U kontrolnojseriji eksperimenata jena isti načindodavan rastvarač (6dodavanjakumulativno, savremenskimrazmakomod15minuta).

Utvrđenojedaterpinolenukocentracijiod11,1%dovodidostatističkiznačajnevazorelaksacije u odnosu na rastvarač kao kontrolu (F=2,584, p<0,05; Two‐wayANOVA). Naši rezultati opravdavaju nastavak ispitivanja potencijalne primeneterpinolenauterapijikardiovaskularnihoboljenja.

Istraživanje je podržalo Ministarstvo prosvete, nauke i tehnološkog razvoja(ProjekatON173021).

709

VASORELAXANTACTIVITYOFTERPINOLENE

ReljaSuručić1,DraganaDrakul2,NadaKovačević3,TatjanaKundaković‐Vasović3

1DepartmentofPharmacognosy,UniversityofBanjaLuka‐FacultyofMedicine,

RepublicofSrpska,2UniversityofEasternSarajevo‐FacultyofMedicine,RepublicofSrpska(BosniaandHerzegovina),3DepartmentofPharmacognosy,

UniversityofBelgrade–FacultyofPharmacy(Serbia) Terpenesrepresentavery importantgroupofchemicalcompoundsbecauseof

their industrial use, aswell asmedical usewhich is possible due to their significantbiological effects. They are one of the largest groups of secondary plantmetabolites.Thenameofthisclassofcompoundscomesfromturpentine(oilofturpentine),aliquidproductofpineoleoresindistillationprocess.

Terpinolene(p‐menth‐2,4(8)‐diene)isacolorlessorpaleyellowcoloredliquidwith aromatic odor. This monocyclic, monoterpenic alkene is widely represented inplanttissuesofvariouspines,pashkans,cannabis,teatree,turmericandparsleyleaves,anditisoftenacomponentofessentialoilsobtainedfromCitrus,Mentha,JuniperusandMyristicaspecies.Itisoneofthemaincomponentsoftheessentialoilofaerialpartsofthe endemic species Seseli gracile Waldst. & Kit. (6.1‐57.5%). Terpinolen possessesconfirmed antiradical, antihyperalgesic and antiedematous activity. Since somestructurally similar monoterpenic compounds have previously shown vasorelaxantactivity, the aim of this study was to investigate the vasorelaxant potential ofterpinolene.

Inthepresentstudy,theeffectsofstandardterpinolene(43905Sigma‐Aldrich)on the isolated renal artery ofWistar ratswere investigated. Three to fivemm longarterial ring tissue segments with preserved endothelium were used. The arterialsegments were placed in water bath in the Krebs‐Ringer solution, at 37° C andcontinuouslyaeratedby95%O2and5%CO2.Contractionsof thebloodvesselswereinduced with 10‐6 M of phenylephrine. The endothelium integrity was confirmedpharmacologically,byusingtheacetylcholine(10‐6M).Afteraccomplishingtonicphaseof the contraction, increasing concentration of terpinolene (0.2 % ‐ 33.3 %,cumulatively)in5%carboxymethylcellulose(CMC)solutionwereaddedtotheorganbath. Each subsequent concentration was added only after the end of effect of thepreviouslyappliedconcentration,i.e.approximately15minutesfromthepreviousone.Inthecontrolseriesofexperiments,asolventwasaddedinthesamemanner.

Itwas foundthatterpinoleneat theconcentrationof11.1%exibitsstatisticallysignificant vasorelaxation in comparison to the solvent control (F = 2.584, p <0.05;Two‐way ANOVA). Our results justify a continuation for the further studies ofterpinolene’spotentialinthetreatmentofcardiovasculardiseases.



710

Arh.farm 2018;68: 710-711 LB-P15

APITERAPIJAUMENOPAUZI

LidijaJerotijević

ApiterapeutskodruštvoSrbijeApiBeo(Srbija)

Apiterapijapredstavljaprimenupčelinjihproizvodasaciljemočuvanjazdravlja

ljudii/ilinjihovoglečenja.Svipčelinjiproizvodiimajuširokuupotrebuutradicionalnojmedicini.Menopauzajefiziološkostanježenekojepredstavljakrajnjenereproduktivnefaze.Domenopauzedolazizbognaglogpadanivoaestrogenauorganizmu.Ovostanječesto jepraćenobrojnimneprijatnimsimptomima:vazomotornanestabilnost,atrofijasluzokože i kože urogenitalnog trakta, promene u skeletnom sistemu, promene ukardiovaskularnom sistemu, psihički problemi, promena kvaliteta kože, estetskepromene. Konvencionalnamedicina i farmakologija kao rešenje ovog problema nudehormonsku supstitucionu terapiju. Ova terapija sa sobom nosi višestruke rizike ineželjene efekte koji često prevazilaze dobit za pacijentkinju. Cilj ovog rada je daizučavanjem dostupne literature potvrdi delotvornost i neškodljivost apiterapije urešavanjuproblemakojiprateovostanje.

U simptomatskoj terapiji menopauze opravdana je upotreba svih pčelinjihproizvoda. Matični mleč i pčelinji polen mogu se koristiti kao kauzalna terapija zaotklanjanjeiublažavanjemenopauzalnihproblema.Matičnimlečsadrži10‐hidroksi‐2‐decensku kiselinu i njene derivate. Ovi molekuli imaju mogućnost vezivanja zaestrogenskereceptore.Pri tomemoguda ispoljeagonistički iliantagonističkiefekatuodnosunahumaniestrogenzavisnoodkoncentracijeestrogenauorganizmu.Ovajradnavodi mehanizme agonističkog i antagonističkog dejstva ovih molekula. U radu jepreporučeninačinupotrebematičnogmleča,doza,navedenisumogućineželjeniefekti,interakcijeikontraindikacije.Pčelinjipolenusvomsastavusadržifitoestrogene.Uraduse obrađujumehanizmi dejstva fitoestrogena, navodi se preporučeni način upotrebepolena, mogući neželjeni efekti do kojih dovodi, interakcije sa lekovima ikontraindikacije.Modulatorniefekatmatičnogmlečaipčelinjegpolenanaestrogenskereceptoreomogućavanjihovuefikasnuineškodljivuupotrebuuterapijimenopauzalnihsimptoma. Kao pomoć u menopauzi apiterapeuti preporučuju ženama upotrebumatičnogmlečaipolenazasebnoiliukombinacijisadrugimpčelinjimproizvodima,kaoikombinacijusafitoterapijom.

711

APITHERAPYINMENOPAUSE

LidijaJerotijević

ApitherapysocietyofSerbiaApiBeo(Serbia)

Apitherapy is the application of bee products with the goal of preserving the

health of people and/or their treatment. All bee products are used in traditionalmedicine.Menopauseisthephysiologicalstateofwomananditrepresentstheendofher reproductivephase.Menopause isdue toa suddendrop inestrogen levels in thebody. This condition is often accompanied by numerous unpleasant symptoms:vasomotorinstability,mucosalandskinatrophyintheurogenitaltract,changesintheskeletalsystem,changesinskinquality,aestheticchanges.Conventionalmedicineandpharmacologyofferhormonreplacement therapyasasolution for thisproblem.Thistherapy carriesmultiple risks and side effects that often outweigh the benefits for apatient. The aim of this paper is to study the available literature to confirm theeffectiveness and harmlessness of apitherapy in solving problems that follow thiscondition.

Allbeeproductscouldbeused insymptomaticmenopausetherapy.Royal jellyand bee pollen can be used as causal therapy for the elimination and alleviation ofmenopausal problems. Royal jelly contains 10‐hydroxy‐2decenoic acid and itsderivatives.Thesemoleculeshavetheabilitybindingtoestrogenreceptors.Indoingso,they may exhibit agonistic or antagonistic effects in relation to human estrogen,depending on the estrogen concentrations in the body.Mechanisms of agonistic andantagonistic actionof thesemolecules areprocessed in zhis paper.Also, it iswrittenhow to use royal jelly, in which dose, possible side effects, drug interactions andcontraindicationsarelistedaswell.Themodulatoryeffectofroyaljellyandbeepollenon estrogen receptors enables their effective and innocuous use in the treatment ofmenopausalsymptoms.Asanaidtomenopause,apitherapyrecommendstowomentheuse of royal jelly, pollen and bee pollen separately or in combinationwith other beeproductsorcombinationswithphytotherapy.

712

Arh.farm 2018;68: 712-713 LB-P16

ANTIMIKROBNAIANTIOKSIDATIVNAAKTIVNOSTMETANOLNOGEKSTRAKTAALCHEMILLAVIRIDIFLORAROTHM.(ROSACEAE)

JelenaRadović1,MarinaMilenković2,TatjanaKundaković‐Vasović1

1Katedrazafarmakognoziju,UniverzitetuBeogradu‐Farmaceutskifakultet,

2Katedrazamikrobiologijuiimunologiju,UniverzitetuBeogradu‐Farmaceutskifakultet(Srbija)

Rod Alchemilla L. (Rosaceae) čine višegodišnje zeljaste biljke koje rastu navlažnim livadama širom Evrope, Azije i Severne Amerike. Do sada, najdetaljnijeistraženavrstajevirak,A.vulgarisL.,čijaseherbananašimprostorimatradicionalnokoristi za lečenje digestivnih i ginekoloških problema. Ova droga sadrži značajnukoličinutanina, istogaposedujeadstrigentno,antioksidativno,antimutagenodejstvoiostvaruje pozitivan efekat na zarastanje rana dokazan in vitro.Međutim, ostale vrsteovogrodasuznatnomanjeispitane.Stoga,ciljovogradaje ispitivanjeantimikrobneiantioksidativneaktivnostmetanolnogekstraktavrsteA.viridifloraRothm.kojadosadanijeistražena.

NadzemnideloviA.viridifloraprikupljenisuujulu2013.godinenaSuvojPlanini(Srbija), nakon čega su osušeni prirodnim putem (320 g), ekstrahovani 70%metanolom,pričemujedobijenmetanolniektrakt(80g).Sadržajukupnihpolifenolaumetanolnom ekstraktu određen je spektrofotometrijski Folin‐Ciocalteu metodom.Sposobnost uklanjanja slobodnih radikala metanolnog ekstrakta je ispitivanastandardnimDPPHtestom,iizraženakaokoncentracijakojadovodidouklanjanja50%DPPHradikala(IC50)ipoređenasastandardnimsupstancama.Antimikrobnaaktivnostmetanolnog ektrakta određena je bujon dilucionom metodom na devet bakterijskihsojeva.

Utvrđeni sadržaj ukupnih polifenola u metanolnom ekstraktu iznosi 205,8 µggalne kiseline/g suvog ektrakta, dok je dobijena IC50 vrednost u DPPH testu bila 7,8µg/ml. Vrednosti IC50 za vitamin C iznosi 3,8 µg/ml i za rutin 5,75 µg/ml.Metanolniekstrakt inhibira rast svih testiranih bakterijskih sojeva pri minimalnoj inhibitornojkoncentraciji (MIC)od125µg/ml,osimbakterijskogsojaSalmonellaabony (MIC62,2µg/ml).

MetanolniekstraktvrsteA.viridifloraispoljavasnažanantioksidativnipotencijal,približan standardnim antioksidansima, koji je u korelaciji sa visokim sadržajemukupnih polifenola u ekstraktu. Međutim, ispitivani ekstrakt pokazuje umerenuantimikrobnuaktivnostnatestiranebakterijskesojeve.


(ProjekatON173021).

713

ANTIMICROBIALANDANTIOXIDANTACTIVITYOFMETHANOLEXTRACTOFALCHEMILLAVIRIDIFLORAROTHM.(ROSACEAE)

JelenaRadović1,MarinaMilenković2,TatjanaKundaković‐Vasović1

1DepartmentofPharmacognosy,UniversityofBelgrade‐FacultyofPharmacy,2DepartmentofMicrobiologyandImmunology,UniversityofBelgrade‐Faculty

ofPharmacy(Serbia)

PlantsfromgenusAlchemillaL.(Rosaceae)areaperennialherbaceousthatgrow

throughoutEurope,AsiaandNorthAmerica.ThemostthoroughlystudiedspeciesistheA.vulgarisL.,whoseherbisusedtraditionallyfordigestiveandgynecologicalproblems.This drug contains tannins, and therefore possesses an astringent, antioxidant,antimutageniceffectandproducesapositiveeffectonwoundhealing.However,otherspecies of this genus are less investigated. Therefore, the aim of this study is toinvestigate the antimicrobial and antioxidant potential of the methanol extract of A.viridifloraRothm.whichhasnotbeenstudiedsofar.

ArealpartsofA.viridiflorawerecollectedinJuly2013atMt.SuvaPlanina,driednaturally, extracted with 70%methanol, where methanol extract was obtained. Thecontent of total polyphenols was determined by spectrophotometric Folin‐Ciocalteumethod.TheabilityoftheextracttoremovefreeradicalswastestedbystandardDPPHtest, and expressed as a concentration that removes50%ofDPPH radical (IC50) andcompared to standard substances. The antimicrobial activitywas determined by thebrothmicrodilutionmethodonninebacterialstrains.

The determined content of the total polyphenolic compounds in themethanolextractwas205.8μgofgallicacid/gofdryextract,whileobtainedIC50inDPPHtestwas7.8 μg/mL. IC50 values for vitamin C were 3.8 μg/ml and for rutin 5.75 μg/mL. Theextract inhibits the growth of all tested bacterial strains with minimum inhibitoryconcentration(MIC)of125μg/mL,apartfromthebacterialstrainofSalmonellaabony(MICof62.2μg/mL).

Methanol extract of A. viridiflora exhibits a strong antioxidant potential,comparablewithstandardantioxidants,whichwascorrelatedwiththehighcontentofpolyphenolsintheextract.However,extractshowsamildantimicrobialactivityagainsttestedbacterialstrains.



714

Arh.farm 2018;68: 714-715 LB-P17

SADRŽAJESTRAGOLAUETARSKOMULJUIINFUZU

HERBEBOSILJKA,OCIMUMBASILICUML.

JelenaArsenijević,AleksandarJovanović,StefanJevtić,NadaKovačević,MilicaDrobac

Katedrazafarmakognoziju,UniverzitetuBeogradu–Farmaceutskifakultet

(Srbija)

Herbabosiljkasetradicionalnokoristikaokarminativ,spazmolitik,blagsedativi

laktagog.Etarskouljeherbebosiljkasadržifenilpropanoidestragol,kojijekancerogenigenotoksičan, a čija količina zavisi od hemotipa bosiljka. Količina estragola koja seunosiputembiljnihlekovitihproizvoda,premapreporuciEvropskeagencijezalekove(EMA), ograničena je na 0,5 mg dnevno tokom 14 dana. Cilj rada bio je ispitivanjehemijskogsastavaetarskihulja,sadržajaestragolauetarskimuljimai infuzimaherbebosiljka,dostupneuSrbiji.

Ispitivani su komercijalni uzorci monokomponentnih čajeva i uzorci herbebosiljka gajenogudomaćinstvimau različitimdelovima Srbije. Infuzi supripremljeniprelivanjem uzoraka ključalom vodom (2 g/150 ml) i ceđenjem nakon 15 min.Izolovanje etarskog ulja iz herbe, kao i izolovanje estragola iz infuza izvršeno jedestilacijom vodenom parom u aparaturi po Klevendžeru. Analiza etarskih ulja idestilatainfuzaizvršenajeGC‐FID‐MSmetodom.Sadržajestragolauetarskimuljimaiinfuzimaodređenjemetodomeksternogstandarda.

Sastav svih ispitivanih etarskih ulja odgovara evropskom hemotipu bosiljka,kojeg karakteriše visok sadržaj linalola ili linalola i estragola. Sadržaj estragola uispitivanim etarskimuljima iznosio je 2,1‐565,4mg/ml, a u infuzima0,5‐11,3 µg/ml.Primenom ispitivanih infuza prema preporuci Deutsche Arzneimittel‐Codex (DAC) (2g/150ml,triputadnevno),dnevniunosestragolaiznosiobi0,2‐5,1mg.

Primenom infuza pojedinih uzoraka herbe bosiljka postoji mogućnostprekoračenjа dozvoljenоg dnevnog unosa estragola. Dobijeni rezultati ukazuju naznačaj utvrđivanja sadržaja estragola u herbi bosiljka koja se koristi u obliku biljnihlekovitihproizvoda.


(ProjekatON173021).

715

THECONTENTOFESTRAGOLEINESSENTIALOILANDINFUSIONOF

BASILHERB,OCIMUMBASILICUML.

JelenaArsenijević,AleksandarJovanović,StefanJevtić,NadaKovačević,MilicaDrobac

DepartmentofPharmacognosy,UniversityofBelgrade–FacultyofPharmacy

(Serbia)

Basil herb is traditionally used as carminative, spasmolytic,mild sedative and

lactagogue. Basil essential oil contains phenylpropanoide estragole, which iscarcinogenicandgenotoxic,anditsamountdependsonBasilchemotype.Theamountof estragole administered through herbal medicinal products, according to therecommendationof theEuropeanMedicinesAgency(EMA), is limited to0.5mgdailyduring 14 days. The aim of this work was examination of chemical composition ofessential oils, as well as determination of estragole content in essential oils andinfusionsofBasilherbavailableinSerbia.

WehaveanalyzedcommercialsamplesofmonocomponentBasilherbalteasandsamplesofBasilherbcultivatedindifferentpartsofSerbia.Infusionswerepreparedbysteeping samples in boilingwater (2 g/150ml) for 15min and subsequent filtering.Isolationof theherbessentialoil, aswellas isolationofestragole from infusionswascarriedoutbyhydrodistillationusingaClevenger‐typeapparatus.AnalysesofessentialoilsandinfusiondistillateswereperformedusingGC‐FID‐MS.Thecontentofestragoleinessentialoilsandinfusionswasdeterminedusingexternalstandardmethod.

ChemicalcompositionofallinvestigatedessentialoilscorrespondstoEuropeanchemotype of Basil that is characterized by high content of linalool or linalool andestragole. The content of estragole in essential oils ranged from2.1 to 565.4mg/ml,whereas infusions contained 0.5‐11.3 µg/ml of this compound. If the examinedinfusionsareadministeredaccordingtotherecommendationofDeutscheArzneimittel‐Codex(DAC)(2g/150ml,threetimesdaily),thedailyintakeofestragolewouldbe0.2‐5.1mg.

ByadministrationofinfusionsofsomeBasilherbsamples,thereisapossibilityofexceedingthepermitteddaily intakeofestragole.Theobtainedresults indicatetheimportanceofdetermining the contentof estragole inBasil herbused in the formofherbalmedicinalproducts.



716

Arh.farm 2018;68: 716-717 LB-P18

ISPITIVANJETRITERPENAUDIHLORMETANSKIMEKSTRAKTIMA

28VRSTARODAHIERACIUML.(ASTERACEAE)SABALKANSKOGPOLUOSTRVA

VioletaMilutinović1,MarjanNiketić2,MilošPetković3,SilvanaPetrović1

1Katedrazafarmakognoziju,UniverzitetuBeogradu‐Farmaceutskifakultet,2Prirodnjačkimuzej,3Katedrazaorganskuhemiju,UniverzitetuBeogradu‐

Farmaceutskifakultet(Srbija)

Rod Hieracium L. s. str. (Asteraceae) predstavlja jedan od najbrojnijih i

najkompleksnijih rodova skrivenosemenica. Na Balkanskom poluostrvu, najveći brojvrsta ovog roda zabeležen je na Dinaridima, naročito na Durmitoru u Crnoj Gori. Ciljovogistraživanjabiojedaseutvrdiimeđusobnouporediprofiltriterpenskihjedinjenjaudihlormetanskimekstraktimanadzemnihdelovaucvetu28vrstaovogrodaporeklomizCrneGoreiSrbije,ito:H.gymnocephalumGriseb.exPant.,H.orieniA.Kern.,H.bleciciiNiketić,H. paratrichum Niketić,H. spirocaule Niketić,H.mokragorae (Nägeli & Peter)Nägeli & Peter, H. pannosum Boiss. s.l., H. plumulosum A. Kern., H. villosum Jacq., H.pilosum Froel.,H. pseudoschenkii (Rohlena & Zahn) Niketić,H. naegelianum Pančić,H.anastrum(Degen&Zahn)Niketić,H.calophyllumR.Uechtr.,H.scheppigianumFreyn,H.durmitoricum (Rohlena & Zahn) Niketić, H. guentheri‐beckii Zahn, H. mirificissimumRohlena & Zahn, H. coloriscapum Rohlena & Zahn, H. pyricephalum Niketić, H.albopellitum(Zahn)Niketić,H.glabratumWilld.,H.scorzonerifoliumVill.s.l.,H.dentatumHoppe s.l.,H. valdepilosum Vill. s.l.,H. neilreichii Beck,H. tommasinianum K. Malý iH.macrodontoides(Zahn)Zahn.

Usuvimdihlormetanskimekstraktimanadzemnihdelovabiljakaucvetuvršenaje komparativna kvalitativna i kvantitativna GC‐FID‐MS analiza triterpena. Strukturatriterpenautvrđenajepoređenjemsakomercijalnimstandardima,kaoinaosnovu1Di2DNMRspektaraodgovarajućihfrakcija.

Uispitivanimvrstamaidentifikovanojeukupnopettriterpena,itodvaderivataursana(α‐amirininjegovacetat),dvaderivataoleana(β‐amirininjegovacetat)ijedanderivatlupana(lupeol‐acetat).Dominantnitriterpeni,uvećiniispitivanihvrsta,bilisuderivatiursana,apraćenisujedinjenjimatipaoleana.

U ovom radu je, za većinu (26) odabranih biljnih vrsta, ispitivanje triterpenasprovedenopoprviput.Naosnovudobijenihrezultata,značajnosuupotpunjenipodacioprisustvuikompozicijitriterpenskihjedinjenjauvrstamarodaHieracium.


(ProjekatON173021).

717

INVESTIGATIONOFTRITERPENESINTHEDICHLOROMETHANE

EXTRACTSOF28HIERACIUML.SPECIES(ASTERACEAE)FROMTHEBALKANPENINSULA

VioletaMilutinović1,MarjanNiketić2,MilošPetković3,SilvanaPetrović1

1DepartmentofPharmacognosy,UniversityofBelgrade‐FacultyofPharmacy,2NaturalHistoryMuseum,3DepartmentofOrganicChemistry,Universityof

Belgrade‐FacultyofPharmacy(Serbia)

ThegenusHieraciumL.s.str.(Asteraceae)representsoneofthemostnumerous

and most complex genera of flowering plants. In the Balkan Peninsula the largestnumber of species of this genus are recorded on Mt Durmitor in Montenegro. Theobjective of this study was to determine and compare triterpene profile of floweringaerial parts dichloromethane extracts of 28 Hieracium species from Montenegro andSerbia: H. gymnocephalum Griseb. ex Pant., H. orieni A. Kern., H. blecicii Niketić, H.paratrichum Niketić, H. spirocaule Niketić, H.mokragorae (Nägeli & Peter) Nägeli &Peter,H.pannosumBoiss.s.l.,H.plumulosumA.Kern.,H.villosumJacq.,H.pilosumFroel.,H.pseudoschenkii(Rohlena&Zahn)Niketić,H.naegelianumPančić,H.anastrum(Degen& Zahn) Niketić, H. calophyllum R. Uechtr., H. scheppigianum Freyn, H. durmitoricum(Rohlena&Zahn)Niketić,H.guentheri‐beckiiZahn,H.mirificissimumRohlena&Zahn,H.coloriscapumRohlena&Zahn,H.pyricephalumNiketić,H.albopellitum(Zahn)Niketić,H.glabratumWilld.,H.scorzonerifoliumVill.s.l.,H.dentatumHoppes.l.,H.valdepilosumVill.s.l.,H.neilreichiiBeck,H.tommasinianumK.MalýandH.macrodontoides(Zahn)Zahn.

In dried dichloromethane extracts of flowering aerial plants parts comparativequalitative and quantitative GC‐FID‐MS analysis of triterpenes was performed. Thestructuresoftriterpeneswereelucidatedcomparingtocommercialstandards,andonthebasisof1Dand2DNMRspectraofcorrespondingfractions.

In investigated species, overall five triterpenes, including two of ursane(α‐amyrinanditsacetate),twoofoleane(β‐amyrinanditsacetate)andoneoflupane(lupeolacetate)type,wereidentified.Dominanttriterpenes,forthemajorityofspecies,wereofursanetype,followedbyrepresentativesofoleanetype.

In this paper, for themajority (26) of selected plant species, triterpenesweretested for the first time. Based on obtained results, the data on the presence andcomposition of triterpen compounds in Hieracium species have been significantlysupplemented.



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