超临界二氧化碳梯度结晶穿心莲内酯的纯化工艺研究

2004,37(12):2047-2050ScientiaAgriculturaSinica12300692230026CO2CO220MPa5590minCO215L·min-180%StudyonPurifyingTechnologyofAndrographolidebySupercriticalCarbonDioxideGradientCrystallizationZHANGWen-cheng1,PANJian1XIEHui-ming1,CHENKe-xun2(1KeyLaboratoryofBioprocessofStateEducationMinistryofHefeiUniversityofTechnology,Hefei230069;2SchoolofChemistryandMaterial,UniversityofScienceandTechnologyofChina,Hefei230026)AbstractEffectsofgradientcrystallizingpressure,temperatureandtimeonthepurityofandrographolidewereinvestigatedviasinglefactorexperiments,andthepurityofandrographolidewasbyHPLC.TheresultshowedthatandrographolidecouldbesynchronouslyextractedandcrystallizedinextractioncontainerbysupercriticalCO2extraction.Moreover,thepurityofandrographolidecrystalwasdistributedgradientlyoncrystallizationboard.Whentheoptimizedparametersofpressure20MPa,temperature55andtime90minwereselected,thecontentofobtaineandrographolidewasmorethan80%.Keywords:Supercriticalcarbondioxide;Gradientcrystallization;Andrographolide;Highperformanceliquidchromatography(HPLC)[Andrographispaniculata(Burm.f.)Nees]andrographolide[12](supercriticalfluidextraction,SCFE)[34]SCFE[5~7]SCFECO2[8]40MPa11.6L101WatersHPLC5152487Rheodyne7725iMillennium3220503730%99.8%31.2.3.4.5.6.7.8.(I)9.(II)10.11.12.1.Gasvase,2.Filter,3.Compressor,4.Liquidmaterialvessel,5.Hotexchanger,6.Extractionandcrystallizationcontainer,7.Crystalware,8.SeparationcontainerI,9.SeparationcontainerII,10.Waterbath,11.Flowmeter,12.BoosterFig.1TheapparatusofsupercriticalCO2extractionandcrystallization1.82mg5ml1.0ml5ml26101418µl[9]WatersSymmetryshield5µmC183.9mm150mm-65%35%1.00ml·min-1205nmµg0.141.31µgY=3472+673X,r=0.99762.00mg25ml[9]30%5590minCO215L·min-115MPa5-2Fig.2Distributionofandrographolidecrystalonthecrystallizationboard812162024MPa3-3Fig.3Effectofgradientcrystallizationpressureonthecontentofandrographolideatthesamepositionofcrystalboard1220491015202530MPa2-1Table1AndrographolidemassoncrystalboardversusdifferentpressureMassofcrystal(g)Relativemassofcrystal%Pressure(MPa)UpsideTotalUpside100.12130.376732.2150.42531.239934.3200.47221.220238.7250.26360.794033.2300.17070.607528.1CO231SC-CO2[10]20MPa80%38.7%-20MPa90minCO215·30%354555653-4-55520MPa55CO215L·min-145607590105120min2-6)Fig.4EffectofgradientcrystallizationtemperatureonthecontentofandrographolideatthesamepositionofcrystalFig.5EffectoftemperatureongradientcrystallizationtimeFig.6Effectoftimeonthecontentofandrographolide675min90min80.0%50g135020MPa,55205037CO2·(7)782.3%A.B.30%C.82.3%A.StandardcontrastofandrographolideB.30%andrographolideC.82.3%drographolideFig.7HPLCgraphsshowingthepurityofandrographolidecrystalCO2CO2·%References[1]DuQZ,JerzG,WinterhalterP.SeparationofandrographolideandneoandrographolidefromtheleavesofAndrographispaniculatausinghigh-speedcounter-currentchromatography.JournalofChromatographyA,2003,984:147-151.[2]JainDC,GuptaMM,SaxenaS,KumarS.LCanalysisofhepatoprotectivediterpenoidsfromAndrographispaniculata.JournalofPharmaceuticalandBiomedicalAnalysis,2000,22:.705-709.[3]LlomartM.P,LorenzoR.A,CelaR.Optimizationofsupercriticalfluidextractionofphenolandcresolsinsoilsamples.JournalofChromatographyA,1996,723:123-134.[4]ChengJS,TangM,ChenYP.Correlationofsolidsolubilityforbiologicalcompoundsinsupercriticalcarbondioxide:comparativestudyusingsolutionmodelandotherapproaches.FluidPhaseEquilibria,2002,197:483-491.[5]..:,2000:9.ZhuZQ.SupercriticalFluidTechnique-PrincipleandApplication.Beijing:ChemicalIndustrialPress,2000:9.(inChinese)[6]HansB,CharlottaT,LenaD,LennartM.DeterminationoffoodconstituentsbasedonSFE:applicationstovitaminsAandEinmeatandmilk.JournalofBiophysicsMethods,2000,43:391-401.[7]RussellT,FaribaD,NeiR.Currentissuesrelatingtoanti-solventmicronisationtechniquesandtheirextensiontoindustrialscales.JournalofSupercriticalFluid,2001,21:159-177.[8]..1220906A.1999-06-30.PanJ,ZhuJZ.SeparationMethodforMaterialCompositionbySupercriticalFluidCrystallization.ChinesePatent:1220906A.1999-06-30.(inChinese)[9]..(),2001,24(6):1083-1086.HuXQPanJZhangWC,ZengQM.ThedeterminationoflactonesconcentrationinandrographlisbyRP-HPLCmethod.JournalofHefeiUniversityofTechnology,2001,24(6):1083-1086.(inChinese)[10]KröberH,TeipelU.Matericalsprocessingwithsupercriticalantisolventprecipitation:processparametersandmorphologyoftartaricacid.JournalofSupercriticalFluids,2002,22:229-235.