简介:目的:建立党参的ISSR-PCR反应体系,为今后利用ISSR标记技术进行党参鉴定及种质遗传多样性分析提供一个标准化程序.方法:采用试剂盒法提取党参基因组DNA为模板,通过单因素实验分析了ISSR-PCR反应体系中MgCl2、dNTPs、引物浓度、TaqDNA聚合酶、模板DNA用量及退火温度对ISSR-PCR扩增的影响.结果:建立了重复性好,分辨率高的ISSR-PCR反应体系,即在25μL反应体系中,含有10×PCRBuffer缓冲液2.5μL,MgCl22.0mmol·L-1,dNTPs0.5mmol·L-1,引物0.4μmol·L-1,TaqDNA聚合酶1.0U,模板DNA为30ng;扩增程序为:94℃预变性5min,然后94℃变性30s,51.7℃退火1min,72℃延伸1.5min,共计35个循环,循环结束后在72℃延伸7min,4℃保存.结论:建立了适用于党参的ISSR-PCR反应体系,为应用ISSR技术鉴定党参种质资源、分子标记辅助选择育种及其遗传多样性研究奠定了基础.
简介:Si-Miao-Wan(SMW),atradiationalChinesemedicinalformulaconsistingofAtractylodisRhizoma,PhellodendriChinensisCortex,CoicisSemen,andAchyranthisBidentataeRadix,hasbeenusedforthetreatmentofgoutandgoutyarthritisformanyyears.Inthepresentstudy,aliquidchromatographyquadrupole-time-of-flightmassspectrometry(LC-Q-TOF/MS)methodwasestablishedtoidentifythemultipleconstituentsofSMWanditsmetabolitesinratbiologicalsamplesafteroraladministration.Atotalof48compoundsinSMW,including21alkaloids,12organicacids,2terpenes,3lactones,2phytosterols,and8othercompounds,weretentativelycharacterizedwiththediagnostic-ionfilteringstrategy.BasedonthediagnosticionsappliedtoidentifycompoundsinSMW,28prototypecompoundsand10metaboliccompoundsweredetectedinthebiologicalsamples.ThiswasthefirstcomprehensivedrugmetabolisminvestigationofSMWinrats.Thedevelopedmethodcouldbeausefulmeansforidentifyingthemulti-componentsinSMWandthemetaboliccomponents.TheresultsmayhelpexplorethepossiblemetabolicprocessesandmechanismofactionforSMWinvivo.
简介:TocharacterizeandidentifymultipleconstituentsinDanhonginjection(DHI),afastultra-highperformanceliquidchromatographycoupledtoelectrosprayionizationquadrupoletime-of-flighttandemmassspectrometry(UHPLC-ESI-QTOF/MS)methodwasestablishedandvalidatedinthepresentstudy.Atotalof63compounds,including33phenolicacids,2C-glycosylquinochalcones,6flavonoidO-glycosides,4iridoidglycosides,6organicacids,5aminoacids,and3nucleosides,wereidentifiedortentativelycharacterized.Inconclusion,theUHPLC-ESI-QTOF/MSmethodisusefulandefficientforin-depthstructuralelucidationofchemicalcompoundsincomplexmatricesofherbalmedicinessuchasDHI.
简介:Thepresentstudywasdesignedtodevelopandvalidateasensitiveandreliableultrahighperformanceliquidchromatographycoupledwithquadrupoletime-of-flightmassspectrometry(UPLC-QTOF/MS)methodtoseparateandidentifythechemicalconstituentsofQixueShuangbuTincture(QXSBT),aclassictraditionalChinesemedicine(TCM)prescription.UndertheoptimizedUPLCandQTOF/MSconditions,56componentsinQXSBT,includingchalcones,triterpenoids,protopanaxatriol,flavonesandflavanoneswereidentifiedandtentativelycharacterizedwithinarunningtimeof42min.Thecomponentswereidentifiedbycomparingtheretentiontimes,accuratemass,andmassspectrometricfragmentationcharacteristicions,andmatchingempiricalmolecularformulawiththatofthepublishedcompounds.Inconclusion,theestablishedUPLC-QTOF/MSmethodwasreliableforarapididentificationofcomplicatedcomponentsintheTCMprescriptions.