BS ISO 11023-2001 Liquorice extracts (Glycyrrhiza glabra L ) - Determination of glycyrrhizic acid content - Methods using high-performance liquid chromatography《甘草提取物 甘草酸含量的测定 高效液相.pdf

1、BRITISH STANDARD BS ISO 11023:1999 Liquorice extracts (Glycyrrhiza glabra L.) Determination of glycyrrhizic acid content Method using high-performance liquid chromatography ICS 71.100.60; 71.040.40 NO COPYING WITHOUT BSI PERMISSION EXCEPT AS PERMITTED BY COPYRIGHT LAWBS ISO 11023:1999 This British S

2、tandard, having been prepared under the direction of the Consumer Products and Services Sector Committee, was published under the authority of the Standards Committee and comes into effect on 15 September 2001 BSI 08-2001 ISBN 0 580 38314 8 National foreword This British Standard reproduces verbatim

3、 ISO 11023:1999 and implements it as the UK national standard. The UK participation in its preparation was entrusted to Technical Committee AW/54, Essential Oils, which has the responsibility to: A list of organizations represented on this committee can be obtained on request to its secretary. Cross

4、-references The British Standards which implement international or European publications referred to in this document may be found in the BSI Standards Catalogue under the section entitled “International Standards Correspondence Index”, or by using the “Find” facility of the BSI Standards Electronic

5、 Catalogue. A British Standard does not purport to include all the necessary provisions of a contract. Users of British Standards are responsible for their correct application. Compliance with a British Standard does not of itself confer immunity from legal obligations. aid enquirers to understand t

6、he text; present to the responsible international/European committee any enquiries on the interpretation, or proposals for change, and keep the UK interests informed; monitor related international and European developments and promulgate them in the UK. Summary of pages This document comprises a fro

7、nt cover, an inside front cover, the ISO title page, the ISO foreword page, pages 1 to 8, an inside back cover and a back cover. The BSI copyright date displayed in this document indicates when the document was last issued. Amendments issued since publication Amd. No. Date CommentsA Referencenumbe r

8、 ISO11023:1999 (E) INTERNATIONAL STANDARD ISO 11023 Firstedition 19991015 Liquoriceextracts( Glycyrrhizaglabra L.) Determinationofglycyrrhizicacid contentMethodusinghighperformance liquidchromatography Extraitsderglisse( Glycyrrhizaglabra L.)Dterminationdelateneuren acideglycyrrhiziqueMthodeparchrom

9、atographieliquidehaute performanceISO11023:1999(E) OSI OSI9991 rllAightsreserved.Unelssotherwisespeceifid,noparotfhtsipubcilatoinamybereproducedorutziliednianyofrmorbyan ymsnae,lecetrinoc .rehsilbupehtmorfgnitirwninoissimreptuohtiw,mliforcimdnagniypocotohpgnidulcni,lacinahcemro noitazidradnatSrofnoi

10、tazinagrOlanoitanretnI 65elatsopesaC 02evneG1121HC dnalreztiwS tenretnI hc.osiosi dnalreztiwSnidetnirP ii Foreword ISO(theInternationalOrganizationforStandardization)isaworldwidefederationofnationalstandardsbodies(ISO memberbodies).TheworkofpreparingInternationalStandardsisnormallycarriedoutthroughI

11、SOtechnical committees.Eachmemberbodyinterestedinasubjectforwhichatechnicalcommitteehasbeenestablishedhas therighttoberepresentedonthatcommittee.Internationalorganizations,governmentalandnongovernmental,in liaisonwithISO,alsotakepartinthework.ISOcollaboratescloselywiththeInternationalElectrotechnica

12、l Commission(IEC)onallmattersofelectrotechnicalstandardization. InternationalStandardsaredraftedinaccordancewiththerulesgivenintheISO/IECDirectives,Part3. DraftInternationalStandardsadoptedbythetechnicalcommitteesarecirculatedtothememberbodiesforvoting. PublicationasanInternationalStandardrequiresap

13、provalbyatleast75%ofthememberbodiescastingavote. InternationalStandardISO11023waspreparedbyTechnicalCommitteeISO/TC54, Essentialoils . AnnexAofthisInternationalStandardisforinformationonly.INTERNATIONALSTANDARD OSI ISO11023:1999(E) 1 Liquoriceextracts( Glycyrrhizaglabra L.)Determinationof glycyrrhiz

14、icacidcontentMethodusinghighperformanceliquid chromatography 1Scope ThisInternationalStandarddescribesamethodfordeterminingtheglycyrrhizicacidcontentofliquoriceextract (Glycyrrhizaglabra L.)byhighperformanceliquidchromatography. Themethodisnotapplicabletoraworgroundliquoriceroot. 2Principle Thesampl

15、eandstandardsolutionsareprepared,thentheglycyrrhizicacidcontentisdeterminedbyhigh performanceliquidchromatographyusingthemethoddescribedinthisInternationalStandard. 3Reagents Useonlyreagentsofrecognizedanalyticalgrade,unlessotherwisespecified. 3.1 Water,HPLCgrade. 3.2 Referencesubstance ,monoammonia

16、calglycyrrhizate(GMA). Ifareferencematerialofguaranteedpurityisnotavailable,itisrecommendedthattheusersofthisInternational Standardcometoanagreementbetweentheinterestedpartiesonthepurityofthereferencesubstance. 3.3Acetonitrile ,HPLCgrade. 3.4 Aceticacid ,analyticalgrade. 3.5 Elutionsolvent(mobilepha

17、se), composedofthefollowing: 38volumesacetonitrile(3.3), 61volumeswater(3.1), 1volumeaceticacid(3.4). Usingthemeasuringcylinder(4.3),preparetheelutionsolventasfollows. Mix1volumeofaceticacidwith61volumesofwater,thenfilterthemixturethroughafilterforaqueoussolvents (4.5). Filter38volumesofacetonitrile

18、throughafilterfororganicsolvents(4.4). Addthefilteredacetonitriletothefilteredwater/aceticacidmixture.Mix,thendegastheelutionsolventintheultra soundcell(4.8.3)oranysuitablesystem. Donotkeepthissolventmorethan48hatambienttemperature.ISO11023:1999(E) OSI 2 4Apparatus Usuallaboratoryapparatusand,inpart

19、icular,thefollowing. 4.1 Pipettes,ofcapacities5mland10ml. 4.2 Volumetricflasks ,ofcapacities50mland100ml. 4.3 Measuringcylinder . 4.4 Filterfororganicsolvents ,0,5 m mporesize. 4.5 Filterforaqueoussolvents ,0,45 m mporesize. 4.6 Oven,capableofbeingmaintainedat105C+ 2C. 4.7 Analyticalbalance ,capable

20、ofweighingtothenearest0,0001g. 4.8 Separationsystem ,asfollows. 4.8.1 Chromatograph,highperformanceliquidphase. 4.8.2 Pumpingsystem ,forobtainingandmaintainingaconstantorprogrammedhighpressureflow. 4.8.3 Solventdegassingsystem ,suchasanultrasoundcelloranysuitablesystem. 4.8.4 Ultravioletdetectionsys

21、tem ,adjustabletoawavelengthof254nm. 4.9 Recorder orintegrator ,ofcompatibleperformancewithalltheapparatus. 4.10 Column,asfollows: material:stainlesssteelorglass; length:10cmto25cm; internaldiameter:0,4cmto0,5cm; stationaryphase:bondedphasesilicawithoctadecylC18derivedfunctionalgroup,maximumparticle

22、size 5 mm; columnefficiency:thenumberrecommendedoftheoreticalplatesis7000to10000. 4.11 Precolumn ,asfollows: material:stainlesssteel; length:10mmor25mm; internaldiameter:2mmor4mm; stationaryphase:bondedphasesilicawithoctadecylC18derivedfunctionalgroupOSI ISO11023:1999(E) 3 5Procedure 5.1 Preparation

23、ofstandardsolutions 5.1.1 Preparationofthe0,5mg/mlsolution Ina100mlvolumetricflask(4.2)weigh,tothenearest10 4 g,50mgofmonoammoniacalglycyrrhizate(3.2). Addtheelutionsolvent(3.5)anddissolvethemonoammoniacalglycyrrhizate(3.2).Makeuptothemarkwitha furtherquantityofelutionsolvent. Filterifnecessarythrou

24、ghtheorganicsolventfilter(4.4). Itisessentialtoprepareanewsolutioneveryday. 5.1.2 Preparationofdilutesolutionscontaining0,05mg/ml,0,075mg/mland0,1mg/ml 5.1.2.1 0,05mg/mlstandardsolution Usingapipette(4.1),transfer5mlofthe0,5mg/mlstandardsolution(5.1.1)toa50mlvolumetricflask(4.2).Make uptothemarkwith

25、theelutionsolvent(3.5). 5.1.2.2 0,075mg/mlstandardsolution Usingapipette(4.1),transfer15mlofthe0,5mg/mlstandardsolution(5.1.1)toa100mlvolumetricflask(4.2). Makeuptothemarkwiththeelutionsolvent(3.5). 5.1.2.3 0,1mg/mlstandardsolution Usingapipette(4.1),tranfer10mlofthe0,5mg/mlstandardsolution(5.1.1)to

26、a50mlvolumetricflask(4.2).Make uptothemarkwiththeelutionsolvent(3.5). Itisessentialtoprepareanewsolutioneveryday. 5.2 Preparationofthesolutionforanalysis 5.2.1 Preparethesolutionforanalysissuchthattheareaoftheglycyrrhizicacidpeakobtainedisbetweenthe peaksfortheothertwostandardsolutions,onthebasisoft

27、herecommendedmassesandvolumesspecifiedin Table1,accordingtotheassumedglycyrrhizicacidcontentofthesample. Table1Examplesofquantitiesandvolumestobeusedaccordingtotheassumedglycyrrhizicacid contentoftheliquoriceextracts Assumed glycyrrhizicacid content Massoftest portion Volumeofflask Volumeofpipetted

28、solution Volumeofflask V 1 V 2 V 3 %m gm lm lm l 3to5 1 50 5 50 5to10 1 100 5 50 10to15 0,50 100 5 50 20to25 0,25 100 5 50ISO11023:1999(E) OSI 4 5.2.2 Weigh,tothenearest10 -4 g,atestportionofmass m 0 intoavolumetricflaskofvolume V 1 ml. Addanappropriatequantityofelutionsolvent(3.5)todissolvethesampl

29、e.Thenmakeuptothemarkwithelution solvent. Usingapipette,transferavolume( V 2 )ofthissolutiontoavolumetricflaskofvolume V 3 ml. Addanappropriatequantityofelutionsolvent(3.5),mixandmakeuptothemarkwithelutionsolvent. Filterthislastsolutionthroughthefilterfororganicsolvents(4.3)priortoinjecting. 5.3 Det

30、erminationofthewaterandvolatilemattercontentofthesample Weigh,tothenearest10 -4 g,atestportion( m 0 )ofexactly2gandplaceinacalibratedwatchglass. Placeitintheoven(4.6)setat105Cfor24h,untilaconstantmassisreached.Coolthenweightheresidue( m 1 ). Thewaterandvolatilemattercontentofthesample, w,expressedas

31、apercentagebymass,isequalto: w mm m = - () % 01 0 100 where m 0 isthemassofthetestportion,ingrams; m 1 isthemassoftheresidueobtained,ingrams. 6Determination 6.1 Inject10 m lofsolution5.1.2.1. 6.2 Inject10 m lofsolution5.1.2.2. 6.3 Inject10 m lofsolution5.1.2.3. 6.4 Inject10 m lofthesolutionforanalys

32、is(5.2.2). 6.5 Carryouttheoperationsdescribedin6.1to6.4twice. NOTE Iftheinjectionanddetectionsystemsallowthis,itispossibletoinjectquantitiesfrom5 m lto10 m l. 6.6 Measuretheareaoftheglycyrrhizicacidpeakonthechromatogramsobtainedforthestandardsolutions(see annexA). 6.7 Plotacurveofglycyrrhizicacidpea

33、kareaagainsttheconcentrationofmonoammoniacalglycyrrhizateinthe standardsolutions. Acalibrationlinepassingthroughtheoriginisobtained. 6.8 Usingthechromatogramforthesamplesolution,measuretheareaoftheglycyrrhizicacidpeak. Fromthecalibrationline,readthemonoammoniacalglycyrrhizateconcentration, c,inmilli

34、gramspermillilitre.OSI ISO11023:1999(E) 5 7 Expressionofresults Theglycyrrhizicacidcontent,w G , expressedasapercentagebymassondrymatter,iscalculatedusingthe formula: w cVV p mV w G () = - 13 02 100 822 100 839 where m 0 isthemassofthetestportion,ingrams; c isthemeanvalueoftheconcentrationsobtained,

35、inmilligramspermillilitre,ofmonoammoniacal glycyrrhizatereadfromthecalibrationline; p isthepurityofthereferencesubstance,inpercent; w isthewaterandvolatilemattercontentofthesampleforanalysis,asapercentagebymass,determinedin 5.3; 822 isthemolarmassofglycyrrhizicacid; 839 isthemolarmassofmonoammoniaca

36、lglycyrrhizate; V 1 , V 2 , V 3 havethesamemeaningsasindicatedinTable1. 8Test report Thetestreportshallspecify: allinformationnecessaryforthecompleteidentificationofthesample; thesamplingmethodused,ifknown; thetestmethodused,withreferencetothisInternationalStandard; alloperatingdetailsnotspecifiedin

37、thisInternationalStandard,orregardedasoptional,togetherwithdetailsof anyincidentswhichmayhaveinfluencedthetestresult(s); thetestresult(s)obtained;or iftherepeatabilityhasbeenchecked,thefinalquotedresultobtained.ISO11023:1999(E) OSI 6 AnnexA (informative) Examplesofchromatograms FigureA.1Chromatogram

38、ofaliquoriceextractOSI ISO11023:1999(E) 7 FigureA.2ChromatogramofthereferencesubstanceISO11023:1999(E) OSI 8 Bibliography 1 ISO8432, EssentialoilsAnalysisbyhighperformanceliquidchromatographyGeneralmethod.BS ISO 11023:1999 BSI 389 Chiswick High Road London W4 4AL BSI British Standards Institution BS

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