气相—质谱联用PolarisQ Basic Training 英文版.pdf

Thermo FisherS C I E N T I F I CThe World Leader in Serving SciencePolarisQPolarisQ Basic TrainingBasic TrainingKe kongThermo Fisher ScientificPolarisPolarisQ QAncestryAncestry1998-GCQ PlusTRACE GCXcalibur Data System1995-GCQExternal Ion Source1996-GCQ PlusMS/MS Standard1999-PolarisHigh Temp.Ion SourceOutline?Basics of Ion Trap Mass Spectrometry?Trapping Injected Ions?Helium buffer gas?Automatic Gain Control(AGC)?Generating a Mass Spectrum?Mass-selective instability?Resonant ejection?Full Scan MS Scan FunctionWhat is a Quadrupole Ion Trap?zoroEntranceEntranceEndcapEndcapExitExitEndcapEndcapRingRingElectrodeElectrode()tV cos(Ion-molecule reactions at low concentrations)Older ion traps used Internal Ionization+FilamentElectronLensIon TrapElectronMultiplierPolarisQuses External Ionization(Neutrals are kept outside the trap)+IonSourceIon Volume(El or Cl)Ions to TrapNeutralsto VacuumFilamentLensesElectronMiltiplierIon Trap15VConversionDynode+IonInjectionIonTrappingMass-selectiveEjectionBasic Steps77001-13839706027 7 0 0 1-1 3 8 39 7 0 6 0 2Stability Diagram0.00.20.40.60.81.01.21.4qz-0.8-0.6-0.4-0.20.00.2azrz0.10.20.30.40.50.60.70.81.00.0.10.20.30.40.50.60.70.80.91.00zoro22o2oz22o2oz)z2rmeU16a)z2r(meV8q+=+=()tcosVU0a0Uz=Stability Line1.00.0qzaz0.9080az/mVqzz=0.908qzMass-selective Ejection of Ions0.0MassMass-selective Instabilityselective InstabilityRamp RF voltage(V)to sequentiallyRamp RF voltage(V)to sequentiallyeject ions from low m/z to high m/z.eject ions from low m/z to high m/z.z/mVqz*Improved Method of Ejection0.908qz0.0MassMass-selectiveselectiveInstability ScanInstability Scan0.902qz0.0MassMass-selectiveselectiveInstability ScanInstability Scanwith Resonantwith ResonantEjectionEjection476 kHzResonant Ejection+-?Improves resolution,especially for high m/z ions?Amplitude(V)ramped with m/z to optimize resolutionVm/z()tcosV()tcosV7 7 0 0 1-1 3 8 39 7 0 6 0 2Helium Buffer GasBuffer GasBuffer GasDamps IonDamps IonMotionMotion77001-13839706027 7 0 0 1-1 3 8 39 7 0 6 0 2HecollisionHeHeHeHe+Helium Buffer GasWithout HeliumWithout HeliumWith HeliumWith Helium?Trap injected ions by removing KE?Damps ions to center of ion trap?Collision gas for MS/MSIncrease sensitivityIncrease resolution and sensitivityIncrease fragmentation efficiencyResults.Purposes of Buffer Gas+zoro+Trapping Injected Ions22o2zmz4eVD=()tcosV+Maximum Number of Ions22o2zmz4eVD=24o2maxmr4V3N=?Determine the number of analyte ions coming from the ion source?Calculate injection time to fill ion trap to capacityAutomatic Gain Control(AGC)Scale intensity dataTimeInjection escanPrTICescanPrTargetAGCTimeInjection=TimeInjection)z/m(I)z/m(I=110100100010000100000110100100010000100000100000010000000Amount of Sample(arbitrary units)Number of Ions Trapped(arbitrary units)Fixed Ion Injection TimeFixed Ion Injection Time10 ms0.1 msSpaceChargeEffectsDYNAMIC RANGE:103250 ms Injection Time5 s Injection TimeVariable Injection TimeInjection Time OptimizedSpace Charge Effects110100100010000100000110100100010000100000100000010000000Amount of Sample(arbitrary units)Number of Ions Trapped(arbitrary units)TimeUnscaled TIC(counts)TimeInjection Time(ms)Unscaled Electrometer DataTimeUnscaled TIC(counts)TimeInjection Time(ms)TimeScaled TIC(counts)Scale IntensityTimeInjection)z/m(I)z/m(I=?Full scan MS?Single ion monitoring(SIM)?Isolating ions of a particular m/z?MS/MS?Resonant excitation to fragment ionsOutlineVoltage Relationship During a Mass Scan(Voltage Relationship During a Mass Scan(QuadrupoleQuadrupole)VR FV+180R F+Vd c-Vd c+1500-1500Complete Mass ScanRF PotentialDC Potential+2500-250m/z77001-1380970608VEject(V)Gate LensMultiplierFull Scan MS Scan FunctionAGC PrescanMass Analysis ScanIon InjectionMass AnalysisIon InjectionMass AnalysisScan TerminologyPrescanMass AnalysisPrescanMass AnalysisPrescanMass Analysis1st Microscan2nd Microscan3rd MicroscanComplete Scan CycleSave DataFull Scan MS1.Inject2.Detecte-e-e-e-e-Injection RF Levels for Full Scan?Two RF levels are used to inject ions.?The high-mass adjust slider bar controls these two levels.?Custom levels can also be chosen(*).0%100%50%25%75%1st Level2nd Level21212821352135283535m/z Range 50-35050-700350-70010-601010600-10004545*Single-Ion Monitoring(SIM)1.Inject2.Isolate3.DetectHow Do We Isolate Ions?0.908qz0.0Ion we wish to isolateIon we wish to isolate7?Ions at different qzvalues oscillate at different frequencies(o)22qzo500 Hzt=8 msIsolation WaveformsqzqzVEject(V)Gate LensMultiplierSIM Scan FunctionAGC PrescanMass Analysis ScanIon InjectionMass AnalysisIon InjectionMass AnalysisIsolateIon IsolationIon IsolationRepeat for each SIM m/zInjection RF Levels for SIM?A single RF level is used to inject ions.?The RF level is chosen automatically.010203040506001002003004005006007008009001000m/zOptimum Ion RFMS/MS1.Inject2.Isolate3.Fragment4.DetectTriple-Stage Quadrupole“Tandem in Space”“Tandem in Time”MSMS-Resonant Excitation0.908qz0.0Time=2-30 msec0.908qz0.0Product ionsProduct ionsenergy0.01.02.03.04.0Excitation Voltage(V)77001-1285970219MS/MS Parameters?Precursor ion?Excitation voltage?Excitation qz?0.225,0.30,or 0.45IntensityPrecursor ion(Product ions)Resonant Excitation qzValue0.908qz0.00.2250.2250.908qz0.00.908qz0.00.300.300.450.45Fragment ionsFragment ionsnot trapped not trapped Product IonProduct Ionm/z Rangem/z RangeFragmentationFragmentationEnergyEnergy1/41/41/31/31/21/2VEject(V)Gate LensMultiplierMS/MS Scan FunctionAGC PrescanMass Analysis ScanIon InjectionMass AnalysisIon InjectionMass AnalysisIsolateIon IsolationIon IsolationResonantExcitationExciteGC/MS SpectrumGC/MS/MS Product Ion SpectrumIsolation ofPrecursor IonMS/MS Example-ChlordaneFragmentPrecursor IonAnalytical Benefits of MS/MS?Increased selectivity?Concentration of analyte too low?Chemical interferences too high(complex matrix)?Structural characterization?Provides information about chemical structure of unknown compounds?Ease of use?Routine GC/MS/MS is as practical as GC/MSPolarisQ Features?Computer interface uses Ethernet Card Can be used with a notebook?CI Reagent Gas is computer controlled for improved stability?New Analyzer with fewer parts.Easy to remove?High Temperature source up to 300C+/-0.5?New inlet valve with seal that can be replaced without breaking vacuum Electrically controlled evacuation Prevents ion removal tool from accidentally being removedInstallation and Tune?Required tools to perform a successful install?Manuals?Dos and Donts?Explanation of the PolarisQ tuneItems Required for a Successful Install?Various tools as noted in the installation manual?Flow Meter?Gloves for handling the trap and associated components?Cleaning abrasive?Cotton tip cleaners?Soap?Tooth Brush?Methanol Optional Items to Insure Success?Ultrasonic bath?Helium Leak detector?TRACE DPFC CalibratorTuning the PolarisQ?Verify auto tune default parameters are selected?Select tune parameters to calibrate?DFC for dampening gas is this configured?Start Tune?View Interpret tune report?Print tune report for future referenceGeneral Tune ParametersFilament default parametersLens default settingsWaveformsEnvironmental Tune ParametersAutomatic TuneAutomatic Tune Page(EI)?Apply Selection To All FlowsThe Variable dampening gas control option allows the operator toselect various dampening gas flow rates.If this option is present,it must be configured in the PolarisQ configuration section.If this selection in the automatic tune EI is selected,the PolarisQ will tune all selected parameters at each dampening gas setting.This will add as much as one hour+to a tune cycle.Automatic Tune(EI)Purpose:To adjust the RF Frequency to match the Trap Center Ring Electrode so reflected RF is minimal.Commonly referred as the RF dip.The RF dip is displayed by opening the diagnostic pageand selecting RF Dip Calibration.Frequency when matched should be above 1030kHz.1.Verify that the source temperature and vacuum are stable.2.Select Tune|Automatic Tune and uncheck all boxes except for the RF Frequency Calibration.3.Select OK.4.Always calibrate the RF frequency at the operating temperature of the source.?RF FrequencyAutomatic Tune(EI)?Multiplier GainPerformed to obtain the optimum S/N ratio.The gain of the dynode and electron multiplier(detector)must be approximately 300,000.During calibration the voltage on the gate lens(lens 2)is ramped from+20 to-30.At around“0”ions begin to pass to the electron multiplier.The system adjust the multiplier voltage until it gets“pulse counts”around 1000 per second.At this point the gain of the detector can be calculated.The results of this calibration is used in determining when the trap has received the correct number of ions.It is recommended to use this calculated voltage for analytical analysis.Increasing the multiplier voltage gain will cause the AGC to store fewer ions thus decreasing system sensitivity.Automatic Tune(EI)?Res.Ejection Amp.A low ac voltage at a frequency of approximately 1/2 the ring electrode RF is applied to the endcap electrodes of the ion trap.This RF serves to improve the ejection of heavier ions from the trap to improve high mass resolution and sensitivity.Automatic Tune(EI)?Mass CalibrationCalibrates the main RF voltage applied to the ring electrode that is required to eject the calibration gas ions(69,131,264,414,and 614)from the trap.In this manner main RF voltages are assigned to all masses in the scan range of the instrument(10-1000 amu).In the EI mode the ions 69,131,264,414,and 614 are used.During Mass Calibration,the rear end cap voltage is set so only those ions selected for tuning are ejected.Mass Calibration is not performed when Res.Ejection Calibrationis selected.Automatic Tune(EI)?WaveformThere are four calibration events:131(1),414 and 131(2)and 414(2).The first two events calibrate ion frequencies to allow ions to be efficiently isolated using waveforms.The second two calibrate ion frequencies so ions can be efficiently resonantly excited for MS/MS.Waveform is calibrated for each available buffer gas setting.Automatic Tune(EI)?Injection RFDetermines the optimum main RF voltage applied to the ring electrode to trap ions of different mass.Once completed,the instrument is able to efficiently trap any m/z ion.This calibration is used in full scan MS,SIM,and MS/MS when Automatic Injection RF mode is selected in Tune.Dos and Dont?Do go over site prep with customer to insure all is ready for installation?Do use gloves when handling the analyzer or its parts?Do pre-condition your column before inserting it into the analyzer?Do re-tighten the column connection at the transfer line after the first column temperature program?Do keep a printed copy of the tune report,Air/Water spectrum and back ground full scan spectrum.?Do use the preloaded sequence table and methods to perform an overnight bake out and to run specs.Dos and Donts?Dont stop column flow without cooling the transfer line firstTypical Sequence Table for Bake out and Spec DataAir Water SpectrumAir Water SpectrumAir Water SpectrumAir Water SpectrumAir Water SpectrumBackground Spectrum