micrOTOF-Q应用培训培训目的:了解ESI-Q-TOF的基本原理掌握仪器的基本操作和维护数据处理软件的使用潘晨松博士布鲁克·道尔顿公司应用技术支持工程师培训安排第一天•基本理论简介及使用中应注意的要点•microTOFControl软件介绍(1)•测样练习(tunemix、小分子、多肽、蛋白质)•microTOFControl软件(2):Calibration;•DataAnalysis软件介绍(1)•练习DataAnalysis软件的使用•学员练习校正及测样第二天•复习及操作考查•HyStarLC/MS•练习小分子混合样品和蛋白/多肽样品测定•DataAnalysis软件(2)•练习处理LC/MS数据第三天•复习及操作考查•软件介绍与练习(LibraryEditor,IsotopePattern,GenerateMolecularFormular,ReportDesigner…..)•练习处理LC/MS数据•样品测定样品测定•答疑、讨论液质联用型高分辨串联质谱仪——电喷雾四级杆飞行时间(ESI-Q-TOF)商品名:micrOTOF-QDryGasHeaterOrthogonalAcceleratorGlassCapillaryCollisionGasSupplyHexapole电喷雾离子源去溶剂系统飞行管检测器反射镜四级杆Q碰撞池产生离子离子传输与聚焦分辨离子IonGenerationIonTransmissionandfocusIonResolutionmicrOTOF-QIonGeneration:AtmosphericPressureIonization(API)AtmosphericPressureIonization-ElectrosprayAtmosphericPressureIonization-ElectrosprayIonization(API-ESI)Ionization(API-ESI)IongenerationforMSanalysis•Nebulization•Desolvation•Coulombexplosions•DesorptionUnderpropersourceconditions,individualionsonlywillenterthecapillary.Anunstablesignalorcapillarycurrentmayindicateaneedforadjustmentofgases,flowrate,orsprayneedle.SeetheUser’sManualforTroubleshootingtips.NebulizerGasSampleDryGasCapillary(4kV)SprayNeedlegroundedDryGasGenerationofIons-NebulizationDropletformationinpresenceofelectricalfieldattheneedletipwithinthespraychamberHVHVGenerationofIons-DesolvationGenerationofIons-IonEvaporationToobtainspectralinformation,ionsneedtobecreatedandintroducedtotheinstrumentforinterrogationMustconsiderflowratesofSample:•Nebulizer•DrygasGenerationofIons-ESIsourceThesampleisintroducedviathenebulizer.TheESIprocessissupportedbynitrogengas.Thedryinggasheatsupthesourceanddriesthespray.Itactsasacountercurrentflowtothenebulizedsampleensuringcompleteevaporationofthesolventinordertopreventdropletsfromenteringtheglasscapillary.•Thedesolvationunitcontainsthedrygasheatingblockandcapillaryhousing.•Theglasscapillaryinterfacestheatmosphericpressureregionofthespraychamberwiththefistvacuumstageoftheionoptics•separatesatmosphericpressurefromfirstvacuumstage•parts:•dryinggasheater•glasscapillaryGenerationofIons–DesolvationunitElectrosprayFactorsAffectingIonizationNeedleset-up•InnerNeedlePosition•NebulizerPressure•NeedleConditionHighvoltageelectrodes•CapillaryVoltagesettings•ConditionofCapillaryandChamberHighVoltageElements•ConditionofInsulatorsSolutionChemistry•FlowRate•SolutionpH•SamplepKa•SolutionConductivityElectrospraySolutionChemistryMobilephasepHhasamajoreffectforanalytesthatareionsinsolution.BasicpH(>7.0;9preferred)fornegativeionsAcidpH(<7.0;5preferred)forpositiveions*ManipulationofpHcanenhanceperformanceforanalytesthatarenotnormallyionizedinsolution.ElectrospraySampleChemistryPositiveIonModeNegativeIonModeBase+acidSampleAcid+baseSampleRNRRRNRHRHAOCORHOCOR:B+H:B+++:-+A-ElectrosprayBuffersChoosebufferscarefullyforTOFinstrumentsElectrosprayBufferSelection:•VolatilebuffersareusedtomodifymobilephasepH.•Maybeaddedinmobilephaseasapost-columnaddition.•Acidicsolutionsfavorpositiveionmode.•Formicacid,0.1-1.0%•Aceticacid,0.1-1.0%•Ammoniumsaltsfav...