电喷雾四级杆飞行时间Qtof理论

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-ElectrosprayIonization(API-ESI)IongenerationforMSanalysis•Nebulization•Desolvation•Coulombexplosions•DesorptionUnderpropersourceconditions,individualionsonlywillenterthecapillary.Anunstablesignalorcapillarycurrentmayindicateaneedforadjustmentofgases,flowrate,orsprayneedle.SeetheUser’sManualforTroubleshootingtips.NebulizerGasSampleDryGasCapillary(4kV)SprayNeedlegroundedDryGasGenerationofIons-NebulizationDropletformationinpresenceofelectricalfieldattheneedletipwithinthespraychamberHVGenerationofIons-DesolvationGenerationofIons-IonEvaporationToobtainspectralinformation,ionsneedtobecreatedandintroducedtotheinstrumentforinterrogationMustconsiderflowratesofSample:•Nebulizer•DrygasGenerationofIons-ESIsourceThesampleisintroducedviathenebulizer.TheESIprocessissupportedbynitrogengas.Thedryinggasheatsupthesourceanddriesthespray.Itactsasacountercurrentflowtothenebulizedsampleensuringcompleteevaporationofthesolventinordertopreventdropletsfromenteringtheglasscapillary.•Thedesolvationunitcontainsthedrygasheatingblockandcapillaryhousing.•Theglasscapillaryinterfacestheatmosphericpressureregionofthespraychamberwiththefistvacuumstageoftheionoptics•separatesatmosphericpressurefromfirstvacuumstage•parts:•dryinggasheater•glasscapillaryGenerationofIons–DesolvationunitElectrosprayFactorsAffectingIonizationNeedleset-up•InnerNeedlePosition•NebulizerPressure•NeedleConditionHighvoltageelectrodes•CapillaryVoltagesettings•ConditionofCapillaryandChamberHighVoltageElements•ConditionofInsulatorsSolutionChemistry•FlowRate•SolutionpH•SamplepKa•SolutionConductivityElectrospraySolutionChemistryMobilephasepHhasamajoreffectforanalytesthatareionsinsolution.BasicpH(7.0;9preferred)fornegativeionsAcidpH(7.0;5preferred)forpositiveions*ManipulationofpHcanenhanceperformanceforanalytesthatarenotnormallyionizedinsolution.ElectrospraySampleChemistryPositiveIonModeNegativeIonModeBase+acidSampleAcid+baseSampleRNRRRNRHRHAOCORHOCOR:B+H:B+++-+A-ElectrosprayBuffersChoosebufferscarefullyforTOFinstrumentsElectrosprayBufferSelection:•VolatilebuffersareusedtomodifymobilephasepH.•Maybeaddedinmobilephaseasapost-columnaddition.•Acidicsolutionsfavorpositiveionmode.•Formicacid,0.1-1.0%•Aceticacid,0.1-1.0%•Ammoniumsaltsfavorproductionofsingleammoniumadducts.•Generalbuffers•Ammoniumacetate•Ammoniumformate•Triethylamine•Othervolatilesolvents(0.5%)•Basicsolutionsfavornegativeionmode.•Ammoniumhydroxide(pH10-11)**TFAcanactasanionsuppressant—avoiduse.APCIAPCILC/MSAdvantages•ComplementarytoAPI-Electrosprayforlesspolaranalytes.•GoodsensitivityforcompoundsofintermediateMWandpolarity.•LesssensitivetosolutionchemistryeffectsthanAPI-ES.•Tolerateshigherflowrateswithoutdecreaseinsensitivity.Disadvantages•Lessusefulforthermallylabilecompounds.•Requiressomecompoundvolatility.ClassicalAPCIprocess:1.Themobilephaseandanalytearenebulized.2.Thedropletsarevaporized.3.Themobilephasemoleculesareionizedbyelectronsfromthecoronadischarge.4.Theanalytemoleculesareionizedbythemobilephaseions.AtmosphericPressureIonizationAtmosphericPressureIonization(API)isafamilyoftechniqueswhichinclude:Electrospray(ESI)–Solventandsamplearenebulizedbyuseofagasstream.–Smalldropletsaredriedandundergoelectrosprayionformation.–Ionsinsolutionaredesolvatedanddesorbedundertheinfluenceofhighpotentialelectrostaticfields.AtmosphericPressureChemicalIonization(APCI)–Solventandsamplearenebulizedandcompletelyvaporizedbyaheater.–IonizationoccursbothduringthevaporizationprocessandasthesolventandsampleenterthecoronaregionoftheAPCIsource.RelativeApplicabilityofLC/MSTechniquesMolecularWeightAnalytePolarityverypolarnonpolar100,00010,0001,000APCIElectrosprayElectrosprayLC/MSAdvantages•Softestionizationavailable•LC/MSinterfacewithbestsensitivity•Extendsmassrangeformultiplychargedanalytes•Workswithawiderangeofmoderatetohighpolaritycompounds•LowmaintenanceDisadvantages•Solutionchemistryinfluencesionizationprocess•Worksonlyformoderatetohighpolarityanalytes•Adductions(otherthanM+H)possiblewithsomeanalytes•Somesensitivitylossathigherflowrates(~1ml/min)DryGasHeaterOrthogonalAcceleratorGlassCapillaryCollisionGasSupplyHexapole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IonGenerationIonTransmissionandfocusIonResolutionIonTransmissionandFocus布鲁克公司独家专利设计：离子漏斗式传输系统离子流方向•离子传输率大大提高，从而提高灵敏度。•增宽传输离子质量范围离子流方向传统的锥式离子传输DCA