DevelopmentandvalidationofanHPLC–MS/MSmethodtodetermineclopidogrelinhumanplasma

摘要： Aquantitativemethodforclopidogrelusingonline-SPEtandemLC–MS/MSwasdevelopedandfullyvalidatedaccordingtothewell-establishedFDAguidelines.Themethodachievesadequatesensitivityforpharmacokineticstudies,withlowerlimitofquantifications(LLOQs)aslowas10pg/mL.ChromatographicseparationswereperformedonreversedphasecolumnsKromasilEternity-2.5-C18-UHPLCforbothmethods.Positiveelectrosprayionizationinmultiplereactionmonitoring(MRM)modewasemployedforsignaldetectionandadeuteratedanalogue(clopidogrel-d_4)wasusedasinternalstandard(IS).Adjustmentsinsamplepreparation,includingintroductionofanonline-SPEsystemprovedtobethemosteffectivemethodtosolvetheanalyteback-conversioninclinicalsamples.Pooledclinicalsamples(twolevels)werepreparedandsuccessfullyusedasreal-samplequalitycontrol(QC)inthevalidationofback-conversiontestingunderdifferentconditions.Theresultshowedthattherealsampleswerestableinroomtemperaturefor24h.Linearity,precision,extractionrecovery,matrixeffectonspikedQCsamplesandstabilitytestsonbothspikedQCsandrealsampleQCsstoredindifferentconditionsmettheacceptancecriteria.Thisonline-SPEmethodwassuccessfullyappliedtoabioequivalencestudyof75mgsingledoseclopidogreltabletsin48healthymalesubjects. ... （共9頁）