利用大孔树脂从葛根中分离纯化总黄酮_英文_刘火安

JournalofChinesePhamraceuticalSciences2006,15(2):121-126121OptmiizationofPreparativeSeparationandPurificationofTotalFlavonoidsfromRadixPuerariaebyMacroporousResinMethodLIUHuo-an1,2,WANGBo-chu1,DAIChuan-yun1*,SHAOZhi-yong3,HECong-lin1,andJIAYun2(1.CollegeofBioengineering,ChongqingUniversity,Chongqing400030,China;2.CollegeofChemistryandBiology,ChongqingPolytechnicCollege,Chongqing400050,China;3.InterdepartmentofGenetics,IowaStateUniversity,Iowa50011,USA)Abstract:AimToscreentheoptimummacroporousresinandconditionsfortheisolationandpurificationofflavonoidsfromRadixPuerariae.MethodsThestaticanddynamicadsorption/desorp-tionmethodswereused,andtheseparationandpurificationprocesswasevaluatedbymeasuringtheconcentrationoftotalflavonoidinthefractionswithUVspectrophotometer.ResultsTheSP70macro-porousresinwasthemosteffectivecomparedwithothermacroporousresins.Theoptimumconditionswerescreened,whichwere0.5gmL-1correspondingtocrudedrugforconcentrationofextract,pH5-6,andappended60timesthevolumeoftheresinbed(BV)withtheadsorptionspeed2BVh-1,andthevolumeofaq.70%(V/V)ethanolaseluantwas5BVwithdesorptionspeed2BVh-1.Bythismethod,thefinalcontentsoftotalflavonoidsexceeded80%.ConclusionTheSP70macroporousresinisthemosteffectiveoneforlarge-scaleisolationandpurificationofflavonoidsfromRadixPuerar-ia,whichmeetsindustrialneeds.Keywords:RadixPuerariae;totalflavonoids;macroporousresin;separation;purificationCLCnumber:R284.2Documentcode:AArticleID:1003-1057(2006)2-121-06IntroductionPuerarialobata(Willd.)Ohwiisaperennialle-guminousplantnativetoeasternAsia.TheextractofRadixPuerariaeisrichinisoflavonesandhasbeenwidelyusedasantipyretic,antidiarrhoetic,diaphoret-ic,andantiemeticagentsintraditionalChinesemedi-cine[1],andhasrecentlybeenusedasapossibleremedyagainstalcoholism[2].Puerarin(daidzein8-C-glucoside),daidzin(daidzein7-O-glucoside),anddaidzeinarethemajorisoflavonoidsinRadixPuerari-ae[3].Inrecentyears,therehasbeenanincreasingin-terestinthestudyofpurifiedactivecomponents.How-ever,thisapproachhasbeenrestrictedprimarilybe-Receiveddate:2005-09-25.Foundationitem:ScienceandTechnologyCommitteeofChongqinginChina(CSTC.2004BB5122).*Correspondingauthor:Tel[phone],[e-mail]parationmethodsusedsofarwerelimitedtosimpletechniquessuchasextractionandprecipitationleadingtolowrecoveries.ThustherecoveryofthemaincomponentofRadixPuerarie,pu-erarin,wasreportedtobeverylow,about1%,al-thoughwithapurityashighas97%[4,5].Traditional-ly,thechromatographicproceduresusedwerebasedontheadsorbentswithlowpuritygrade,incombinationwithotherunitoperationssuchasfiltrationandextrac-tion[6].Thisiswhyamoreefficientandsimplepurifi-cationmethodforpuerarinisrequired.Macroporousresinisaporoushighmolecularma-terial,whichhasporosityandbiggishspecificsurfacearea.Forseparationandpurificationofnaturalcom-pounds,itadsorbsorganicsubstanceselectivelyfromsolutionthroughphysicalaction.Thepurificationmethodofmacroporousresinisanewpromisingtech-nologyinpharmaceuticalindustry,andhasbeensuc-122JournalofChinesePharmaceuticalSciences2006,15(2):121-126cessfullyappliedintheextraction,separationandpuri-A=0.0957+70.92C(r=0.9999).Samplewasficationofflavonoids,saponinsandalkaloids[7,8].Thequantifiedbyusingthesamemethods.resultsofsomeinvestigations[9,10]showedthatmacro-PreparationofcrudedrugsolutionporousresinAB-8canbeusedtoseparateandpurifythetotalflavonoidfromtheextractsofRadixPueraria.ThepowderedRadixPuerariae(500g)wasex-tracted9timeswith(WV/)30%aqueousethanolun-ThepresentreportshowsSP70toisolateandpurifythederrefluxfor1h×3toaffordextractsolutionsandwastotalflavonoidsfromtheextractsofRadixPuerariawassuperiortoAB-8andhassomeothermeritssuchassimpletechniquesandlowcos.tMaterialsandMethodsMaterialsandinstrumentsRadixPuerariaewasobtainedfromWukuaishiMedicinalMaterialsMarketofChengdu,China(iden-thenfiltered.Thefiltrateeachtimewascollected,con-centratedto0.5gmL-1crudedrug,andthencentrifugedat12000rmin-1for15min.Thesupernatantwasquantifiedfortotalflavonoids,whosecontentwas47.0mgmL-1.ResultsandDiscussionsScreeningofmacroporousresintifiedbyPro.fJiaMR,ChineseMedicineUniversityofChengdu,China).PuerarinwaspurchasedfromNa-Staticadsorption/desorptionThirtymillilitresofcrudedrugsolutionwasputinto10gofpretreatedtionalInstituteforControlofPharmaceuticalandBio-macroporousresinandshaked5minonetimefor2h,logicalProducts,Beijing,China(No:752-200108).Allreagentswereofanalyticalgradeandobtainedfromandtheresinwasthenfiltered.Thefiltratewascollect-edandtheresinwasdesorbedwith30mLof70%a-BeijingChemicalsIndustry,Beijing,China.Macro-porousresinwaspurchasedfromMitsubishiCo.Ltd.ofJapanforSP825,SP850,SP207,SP700,andSP70,andfromChengguangAcademyofChemicalIn-queousethanol.ThecontentoftotalflavonoidsofRa-dixPuerariaeinfiltratesafterbothadsorptionanddep-sorptionweredetermined.Resultsshowed.(Figure1)thatthereare8kindsofresin,AB-8,X-5,HP20,dustry,Chengdu,China,forD4020,AB-8,X-5,D141,D140,andD100,andfromNankaiUniversity,Tianjin,ChinaforHP20,HP2MG,HP2MGL,NKA-9,andNKA-12.Theultravioletabsorptionmeasurementswerecar-r