QTrap-GuidePowerPoint

TMGoStraighttotheAnswers…ThispresentationisintendedasaselfstudyguidetothenewscanfeaturesavailableontheQTrapTM.The“Enhanced”ScanTypes•Introducing6newscantypes:–EnhancedMS(EMS)–EnhancedMultiplyCharged(EMC)–EnhancedResolution(ER)–EnhancedProductIon(EPI)–TimeDelayedFragmentation(TDF)–MS/MS/MS(MS3)EnhancedMS(EMS)-theenhancedsingleMSscan,usedasasurveyscanEnhancedMS(EMS)Q0Q1Q2Q3/LITNoFragmentationRFOnlyTrap&Scan1.IonsarepassedthroughQ1andQ2withoutfragmentation2.Ionsarecollected(trapped)intheLITforaspecifiedtime3.IonsarescannedoutoftheLITanddetected.3+2+EnhancedMS500fmol/µLAngiotensinISampleExperiment-EMSRequiredMaterials:•ReninSubstrateprovidedininstallchemicalkit(suggestedconcentration5pmol/uL)•100uLsyringeprovidedininstallkit•TurboIonSpraySourceEMSRenin,5pmol/uL123451.Fillthesyringewithreninsubstrateandinfuseatarateof5-10uL/min.2.SelectEnhancedMS(EMS)fromthedrop-downscantypelist3.Setthesourceconditionsasdisplayedinthefigureabove(CUR10,CADHigh,IS5000,TEM0,GS120,GS20,iheON)4.Setthemassrangefrom300-1200amu5.Setthenumberofscanstosumto1EMSRenin,5pmol/uL1231.SettheDPbetween20and50V2.ThedefaultEPof10Visgood3.SettheCEto10eVHint-ifyoursampleisfragile(I.e.fragmentseasily),decreasetheCEto5eV.ThiswillminimizefragmentationBUTwillalsoaffectyoursensitivity.44.LeaveIE1atthesetdefaultEMSRenin,5pmol/uL123451.ScaninProfileMode2.Chooseascanspeedwhichwillautomaticallydeterminethestepsize.ForEMS,chooseeither1000or4000amu/s3.Q0TrappingshouldgenerallynotbeusedwithEMSscansinordertopreventpossiblespacecharging.InIDAsituations,youmaychoosetouseQ0trappingtoincreasethedutycycleforyourdependentscan.4.Forbestpeakshapeandresolution,usealowLITfilltime-between5and20ms.5.ThedefaultQ3Entrybarrierof8Visappropriate.Hint-ifyoursampleisfragile(i.e.fragmentseasily),decreasetheQ3EntryBarrierto5eV.ThiswillminimizefragmentationBUTwillalsoaffectyoursensitivity.EMSRenin,5pmol/uLPressStarttoacquireamassspectrum.Thepeaksm/z880,587and436aretheReninSubstrate2+,3+and4+ionsrespectively.EnhancedMultiply-ChargedScan(EMC)-reducingtheamountofsinglychargedionsinyoursingleMSscanEnhancedMultiplyCharged(EMC)Q0Q1Q2Q3/LITNoFragmentationRFOnlyTrapEmpty&ScanQ0Trapping1.IonsarepassedthroughQ1andQ2withoutfragmentation2.Ionsaretrappedinthelineariontrap3.Singlychargedionsareremoved4.RemainingMulitply-ChargedIonsarescannedoutanddetected5.Asionsarebeingscannedoutofthetrap,incomingionsarestoredintheQ0region(Q0trapping)3+2+EnhancedMultiplyCharged500fmol/µLAngiotensinISampleExperiment-EMCRequiredMaterials:•ReninSubstrateprovidedininstallchemicalkit(suggestedconcentration5pmol/uL)•100uLsyringeprovidedininstallkit•TurboIonSpraySourceEMCRenin,5pmol/uL123451.Fillthesyringewithreninsubstrateandinfuseatarateof5-10uL/min.2.SelectEnhancedMulti-Charge(EMC)fromthedrop-downscantypelist3.Setthesourceconditionsasdisplayedinthefigureabove(CUR10,CADHigh,IS5000,TEM0,GS120,GS20,iheON)4.Setthemassrangefrom300-1200amu5.Setthenumberofscanstosumto1EMCRenin,5pmol/uL1231.SettheDPbetween20and50V2.ThedefaultEPof10Visgood3.SettheCEto10eVHint-ifyoursampleisfragile(I.e.fragmentseasily),decreasetheCEto5eV.ThiswillminimizefragmentationBUTwillalsoaffectyoursensitivity.44.LeaveIE1atthesetdefaultEMCRenin,5pmol/uL1.ScaninProfileMode2.Chooseascanspeedwhichwillautomaticallydeterminethestepsize.ForEMC,chooseeither1000or4000amu/s3.UseQ0trapping.4.UseaLITfilltimebetween20and50ms.5.ThedefaultQ3Entrybarrierof8Visappropriate.6.Useafilltimeof50-100ms.123456EMCRenin,5pmol/uLPressStarttoacquireamassspectrum.Thepeaksm/z880,587and436aretheReninSubstrate2+,3+and4+ionsrespectively.Notethatthemultiply-chargedpeaksarethedominantpeaksinthespectrum.Thelowmasssingly-chargedionsareminimalEMCRenin,5pmol/uLEMCEMS2+3+4+2+3+4+EnhancedResolutionScan(ER)-theconfirmationscan,usedtodeterminechargestates,isotopepatternsandconfirmmassassignmentEnhancedResolution(ER)Q0Q1Q2Q3/LITNoFragmentationResolvingTrap&ScanQ0Trapping1.IonsarefilteredthroughQ1(Openresolution,5-7amu)2.IonspassthroughQ2withoutfragmentation3.Ionsaretrappedinthelineariontrap4.Ionsarescannedslowlyoutofthetrapanddetected5.Asionsarebeingscannedoutofthetrap,incomingionsarestoredintheQ0region(Q0trapping)EnhancedResolutionAngiotensinI3+2+SampleExperiment-ERRequiredMaterials:•ReninSubstrateprovidedininstallchemicalkit(suggestedconcentration5pmol/uL)•100uLsyringeprovidedininstallkit•TurboIonSpraySourceERRenin,5pmol/uL123451.Fillthesyringewithreninsubstrateandinfuseatarateof5-10uL/min.2.SelectEnhancedResolution(ER)fromthedrop-downscantypelist3.Setthesourceconditionsasdisplayedinthefigureabove(CUR10,CADHigh,IS5000,TEM0,GS120,GS20,iheON)4.Setthecentermassto436,587and880tomonitorthe4+,3+and2+chargestatesrespectively5.Setthenumberofscanstosumto1ERRenin,5pmol/uL1231.SettheDPbetween20and50V2.ThedefaultEPof10Visgood3.TheCEdefaultsto10eVandisnotuseraccessibleHint-ifyoursampleisfragile(I.e.fragmentseasily),decreasetheCEto5eV.ThiswillminimizefragmentationBUTwillalsoaffectyoursensitivity.Todothis,clicktherightmousebuttononthemasstable.Fromthepop-upwindow,chooseCE.CEwillnowbeuser-editableinthemasswindow