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2September1,2008©2008AppliedBiosystemsInc.andMDSInc.JointOwnersABOVE&BEYOND4000QTRAP®LC/MS/MSSystem3200QTRAP®LC/MS/MSSystemQSTAR®ELITELC/MS/MSSystemAPI4000™LC/MS/MSSystemAPI2000™LC/MS/MSSystemAPI3200™LC/MS/MSSystemQTRAP™5500LC/MS/MSSystemTripleQuad™5500LC/MS/MSSystemVersion2.0DiscoveryDiscoveryLeadIDLeadIDLeadOptimizationLeadOptimizationPrePre--clinicalclinicalClinicalClinicalDevelopmentDevelopmentAPI5000™LC/MS/MSSystem3September1,2008©2008AppliedBiosystemsInc.andMDSInc.JointOwnersABOVE&BEYOND4000QTRAP®QTRAP®:-3200QTRAP®QTRAP55004September1,2008©2008AppliedBiosystemsInc.andMDSInc.JointOwnersABOVE&BEYOND3200QTRAP4000QTRAP-1msQQQQqLIT-MS/MS↑500xQQQQ35September1,2008©2008AppliedBiosystemsInc.andMDSInc.JointOwnersABOVE&BEYOND•MRM•••••SIMQtrap•(EPI)•MS3•(EMS)•(EMC)••(TDF)•MRM-EPI•MRM-MS3•-EPI•-EPI•EMS–EPI•……6September1,2008©2008AppliedBiosystemsInc.andMDSInc.JointOwnersABOVE&BEYOND…MRMÆEnhancedProductIon(EPI)MultipleMRM’sIDACriteriaLevel1EPIAddtoExclusionList•MRMIDA:•MRM•LINAC(~300)•MRMEPIMS/MS•MRMIDA(InformationDependentAcquisition)7September1,2008©2008AppliedBiosystemsInc.andMDSInc.JointOwnersABOVE&BEYONDMRM-IDA-EPI-TICof-MRM(2pairs):Exp1,fromSample5(OA-1)ofOA_EPI.wiff(TurboSpray)Max.6.5e4cps.0.51.01.52.02.53.03.54.04.55.05.56.06.57.07.58.0Time,min0.01.0e42.0e43.0e44.0e45.0e46.0e46.5e43.91TICof-EPI:Exp2,fromSample5(OA-1)ofOA_EPI.wiff(TurboSpray)Max.1.2e6cps.0.51.01.52.02.53.03.54.04.55.05.56.06.57.07.58.0Time,min0.001.00e52.00e53.00e54.00e55.00e56.00e57.00e58.00e59.00e51.00e61.10e61.18e63.95-EPI:Exp2,3.774to4.103minfromSample7(OA-3)ofOA_EPI.wiff(TurboSpray)Max.2.4e4cps.100150200250300350400450500550600650700750800850900m/z,Da0.01000.02000.03000.04000.05000.06000.07000.08000.09000.01.0e41.1e41.2e41.3e41.4e41.5e41.6e41.7e41.8e41.9e42.0e42.1e42.2e42.3e42.4e4803.7255.2563.4293.3151.1321.3191.3785.5121.2545.4303.3133.0167.2125.0343.4219.2161.0307.3643.5385.3421.4457.4477.3237.1739.6MRMEPIMS/MSofOA8September1,2008©2008AppliedBiosystemsInc.andMDSInc.JointOwnersABOVE&BEYOND●MRM334--117334—145●MRM-IDA-EPI(QTRAP)9September1,2008©2008AppliedBiosystemsInc.andMDSInc.JointOwnersABOVE&BEYOND•0.034mg/kg•0.01mg/kg•=0.850•5.78min•=0.909•5.845.87min246Time,min5.7865.8410September1,2008©2008AppliedBiosystemsInc.andMDSInc.JointOwnersABOVE&BEYONDEPI80100120140160180200m/z,amu10%20%30%40%50%60%70%80%90%100%Rel.Int.(%)14511717114711910520013211814691172Solvent-matchedstandardofTebufenpyrad80100120140160180200m/z,amu10%20%30%40%50%60%70%80%90%100%Rel.Int.(%)15117719114613616012120518917611715114710716191Gingerextract11September1,2008©2008AppliedBiosystemsInc.andMDSInc.JointOwnersABOVE&BEYOND●●LC/MS/MSMRM,QTRAP,MRM.12September1,2008©2008AppliedBiosystemsInc.andMDSInc.JointOwnersABOVE&BEYONDXICof+MRM(2pairs):Exp1,277.1/203.0DaID:Clenbuterol_2fromSample5(STD_0.2pg_IDA)of04192009.wiff(TurboSpray)Max.8.8e4cps.1.02.03.04.05.06.07.08.09.010.011.012.0Time,min0.05.0e48.8e44.50XICof+MRM(2pairs):Exp1,286.1/204.2DaID:ISfromSample5(STD_0.2pg_IDA)of04192009.wiff(TurboSpray)Max.1.4e5cps.1.02.03.04.05.06.07.08.09.010.011.012.0Time,min0.05.0e41.0e51.4e54.48+EPI(277.10)Charge(+0)CE(40)CES(15)FT(238.095):Exp2,4.481minfromSample5(STD_0.2pg_IDA)of04192009.wiff(TurboSpray)Max.9.8e4cps.6080100120140160180200220240260280300320m/z,Da0.05.0e49.8e4203.2168.2132.1277.4235.4149.2259.3159.2+EPI(286.10)Charge(+0)CE(40)CES(15)FT(87.108):Exp3,4.492minfromSample5(STD_0.2pg_IDA)of04192009.wiff(TurboSpray)Max.3.6e5cps.6080100120140160180200220240260280300320m/z,Da0.03.6e5204.1169.1133.1286.3268.2241.3141.1199.1227.3185.0136.9152.0181.0215.3111.1245.0105.166.0MRMEPIEPI289/204m/zEPI277/203m/z13September1,2008©2008AppliedBiosystemsInc.andMDSInc.JointOwnersABOVE&BEYONDXICof+MRM(2pairs):Exp1,277.1/203.0DaID:Clenbuterol_2fromSample11(Sample_006)of04192009.wiff(TurboSpray)Max.6.8e4cps.1.02.03.04.05.06.07.08.09.010.011.012.0Time,min0.05.0e46.8e44.347.71XICof+MRM(2pairs):Exp1,286.1/204.2DaID:ISfromSample11(Sample_006)of04192009.wiff(TurboSpray)Max.9.8e4cps.1.02.03.04.05.06.07.08.09.010.011.012.0Time,min0.05.0e49.8e44.404.03+EPI(277.10)Charge(+0)CE(40)CES(15)FT(3.63694):Exp2,4.324minfromSample11(Sample_006)of04192009.wiff(TurboSpray)Max.7.7e6cps.6080100120140160180200220240260280300320m/z,Da0.05.0e67.7e6130.184.070.0277.4120.4105.8186.0180.2236.3125.273.9260.3+EPI(286.10)Charge(+0)CE(40)CES(15)FT(36.8189):Exp2,4.397minfromSample11(Sample_006)of04192009.wiff(TurboSpray)Max.6.5e5cps.6080100120140160180200220240260280300320m/z,Da0.05.0e56.5e5286.3204.1133.2169.0268.3105.080.9200.3137.0115.2240.170.0212.2187.1147.2100.8263.5273.5MRMEPIEPI289/204m/zEPI277/203m/z14September1,2008©2008AppliedBiosystemsInc.andMDSInc.JointOwnersABOVE&BEYOND(4.5min)EPI4.5+EPI(277.10)Charge(+0)CE(40)CES(15)FT(238.095):Exp2,4.481minfromSample5(STD_0.2pg_IDA)of04192009.wiff(TurboSpray)Max.9.8e4cps.6080100120140160180200220240260280300320m/z,Da0.02.0e44.0e46.0e48.0e49.8e4203.2168.2132.1277.4235.4167.2149.2159.2195.1175.1151.3217.885.0259.1133.2213.0124.1103.898.9232.3241.074.0+EPI(286.10)Charge(+0)CE(40)CES(