Agilent 6510 Q-TOF LC/MS Techniques and Operation

仪器资料 · 发表于 2018-4-26 13:20:46

Table Of Contents
Q-TOF THEORY ...........................................................................................................................1
IN THIS SECTION ...........................................................................................................................2
MASS ANALYSIS ...........................................................................................................................3
MASS ANALYZER PERFORMANCE CHARACTERISTICS...................................................................5
EXACT MASS ..............................................................................................................................13
MASS ACCURACY .......................................................................................................................14
PERFORMANCE CHARACTERISTICS - SUMMARIZED.....................................................................17
QUADRUPOLE OPERATION..........................................................................................................18
TANDEM MASS ANALYSIS ..........................................................................................................25
COLLISION INDUCED DISSOCIATION ...........................................................................................27
TANDEM MASS ANALYSIS ..........................................................................................................29
Q-TOF........................................................................................................................................30
ADVANTAGES OF Q-TOF............................................................................................................31
Q-TOF OPERATIONAL MODES....................................................................................................32
TIME-OF-FLIGHT.........................................................................................................................36
VARIATIONS IN FLIGHT TIME......................................................................................................39
VARIATIONS IN KINETIC ENERGY ...............................................................................................40
STABLE FLIGHT TIMES................................................................................................................41
TIME TO MASS CONVERSION ......................................................................................................43
TIME TO MASS CALIBRATION .....................................................................................................44
TDC VERSUS ADC .....................................................................................................................45
AGILENT 6510 Q-TOF HARDWARE OVERVIEW...............................................................53
IN THIS SECTION .........................................................................................................................54
Q-TOF APPLICATIONS................................................................................................................55
Q-TOF SPECIFICATIONS .............................................................................................................57
Q-TOF SYSTEM DIAGRAM .........................................................................................................59
Q-TOF ION SOURCES..................................................................................................................61
Q-TOF ION OPTICS.....................................................................................................................63
QUADRUPOLE .............................................................................................................................71
COLLISION CELL.........................................................................................................................72
SLICER ASSEMBLY......................................................................................................................78
FLIGHT ASSEMBLY .....................................................................................................................79
PULSER ASSEMBLY.....................................................................................................................81
MIRROR ASSEMBLY....................................................................................................................82
DETECTOR ASSEMBLY................................................................................................................83
VACUUM SYSTEM.......................................................................................................................85
MASSHUNTER WORKSTATION ...................................................................................................91
IONIZATION AND SOURCES ..................................................................................................93
IN THIS SECTION .........................................................................................................................94
Q-TOF ION SOURCES..................................................................................................................95
LC/MS TECHNIQUES ..................................................................................................................96
ELECTROSPRAY ..........................................................................................................................98
APCI ........................................................................................................................................109
MULTIMODE .............................................................................................................................116
APPI.........................................................................................................................................124
CHIP CUBE................................................................................................................................129
PDF-MALDI............................................................................................................................132
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LABORATORY ...........................................................................................................................138
AGILENT Q-TOF CALIBRATION AND TUNING ..............................................................139
IN THIS SECTION .......................................................................................................................140
CALIBRATION AND TUNING ......................................................................................................141
CALIBRANT AND TUNING SOLUTION INTRODUCTION ...............................................................142
Q-TOF TUNE SOLUTION - ESI ..................................................................................................143
Q-TOF TUNE SOLUTION – APCI ..............................................................................................144
Q-TOF TUNE SOLUTION – MMI...............................................................................................145
SOLVENT SELECTION VALVE PLUMBING ..................................................................................146
ENTERING CALIBRATION AND TUNING .....................................................................................147
BEST RESULT: CALIBRATION AND TUNING..............................................................................148
CALIBRATIONS PRIOR TO A RUN...............................................................................................149
CALIBRATION RESULT- MASS CALIBRATION CURVE................................................................150
CALIBRATION RESULT – DETAILED RESIDUAL PLOT ................................................................151
AUTOMATIC AND MANUAL TUNING .........................................................................................153
ORDER OF TUNING CHOICES .....................................................................................................154
AUTOTUNES – CHECK TUNE .....................................................................................................155
CHECK TUNE REPORT...............................................................................................................156
AUTOTUNES – QUICK TOF TUNE..............................................................................................157
AUTOTUNES – TOF AND QUAD AUTOTUNE..............................................................................158
AUTOTUNE REPORT ..................................................................................................................160
INITIAL TUNES ..........................................................................................................................161
SET DETECTOR GAIN ................................................................................................................163
Q-TOF TUNE FILE STRUCTURE ................................................................................................164
SAVE TUNE FILE WHEN CHANGING CONTEXT..........................................................................165
MANUAL TUNING .....................................................................................................................166
CREATE AND SAVE A NEW MASS LIST IF DESIRED ...................................................................167
MANUAL TUNE – FIRST STEPS ..................................................................................................168
MANUAL TUNING – RAMP PARAMETERS ..................................................................................170
MANUAL TUNE – RAMP PARAMETERS......................................................................................174
HOW DO QUAD WIDTH GAIN AND OFFSET AFFECT PEAK WIDTHS?.........................................176
SUGGESTED OPTIMIZATION ......................................................................................................178
RECALIBRATE FOR LOW MASS RANGE APPLICATIONS .............................................................180
LABORATORY ...........................................................................................................................182
LABORATORY EXERCISE: Q-TOF CALIBRATION AND TUNING.............................183
IN THIS LABORATORY EXERCISE, YOU WILL NEED: .................................................................184
ENTER TUNE CONTEXT.............................................................................................................185
TOF CALIBRATION ...................................................................................................................186
AUTOTUNES..............................................................................................................................188
MANUAL TUNING .....................................................................................................................189
IMPORTANT HPLC TIPS AND MAINTENANCE...............................................................191
IN THIS SECTION .......................................................................................................................192
1200 STACK..............................................................................................................................193
FITTINGS...................................................................................................................................195
CAPILLARIES.............................................................................................................................197
EXTRA-COLUMN VOLUME........................................................................................................198
SOLVENT RECOMMENDATIONS .................................................................................................199
CLEANING SOLVENT RESERVOIRS ............................................................................................204
CLEANING THE HPLC...............................................................................................................205
SOLVENT MISCIBILITY..............................................................................................................207
MOBILE PHASE PREPARATION ..................................................................................................208
KEYS TO SUCCESS.....................................................................................................................210
SOLVENT COMPRESSIBILITY .....................................................................................................211
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PRIMING THE HPLC..................................................................................................................213
SAMPLE PREPARATION .............................................................................................................214
COLUMNS .................................................................................................................................220
1200 ROUTINE MAINTENANCE..................................................................................................223
1200 TESTS...............................................................................................................................225
PUMP MAINTENANCE................................................................................................................226
1200 HPLC DIAGNOSTIC SOFTWARE .......................................................................................231
EVALUATE PUMP ......................................................................................................................232
HIGH PERFORMANCE AUTOSAMPLER .......................................................................................233
THERMOSTATTED COLUMN COMPARTMENT.............................................................................236
COLUMN SWITCHING VALVE....................................................................................................238
PRESSURE PROBLEMS ...............................................................................................................240
LABORATORY ...........................................................................................................................244
METHODS AND COLUMNS FOR LC/MS ............................................................................245
IN THIS SECTION .......................................................................................................................246
CONVERTING METHODS ...........................................................................................................247
MOBILE PHASE SELECTION.......................................................................................................249
COLUMNS – BONDED PHASES ...................................................................................................252
COLUMN CONFIGURATION - ID.................................................................................................257
COLUMN CONFIGURATION – PARTICLE SIZE.............................................................................258
RAPID RESOLUTION COLUMNS .................................................................................................263
RAPID RESOLUTION METHOD TRANSFER..................................................................................269
RAPID RESOLUTION INSTRUMENTATION...................................................................................271
COLLISION INDUCED DISSOCIATION (CID) ...................................................................277
IN THIS SECTION .......................................................................................................................278
CID...........................................................................................................................................279
INTERPRETATION OF CID SPECTRA...........................................................................................281
SIGHT OF IONIZATION ...............................................................................................................283
COMMON NEUTRAL LOSS .........................................................................................................284
ION STABILITY..........................................................................................................................285
FRAGMENTATION MECHANISMS...............................................................................................286
SINGLE BOND CLEAVAGE .........................................................................................................287
CYCLIZATIONS..........................................................................................................................288
MULTIPLE CLEAVAGES.............................................................................................................289
RING OPENING..........................................................................................................................290
PEPTIDE SEQUENCING...............................................................................................................291
CHARGE REMOTE FRAGMENTATION.........................................................................................292
FRAGMENTATION EXAMPLE .....................................................................................................294
Q-TOF ACQUISITION..............................................................................................................299
IN THIS SECTION .......................................................................................................................300
HPLC CONTROL.......................................................................................................................301
OPEN DATA ACQUISITION.........................................................................................................302
DATA ACQUISITION SOFTWARE................................................................................................303
INSTRUMENT STATUS................................................................................................................304
INSTRUMENT CONTROL ............................................................................................................306
REAL-TIME PLOT ......................................................................................................................307
SINGLE SAMPLE METHOD.........................................................................................................309
METHOD EDIT...........................................................................................................................310
HPLC PARAMETERS .................................................................................................................311
Q-TOF PARAMETERS................................................................................................................318
Q-TOF ACQUISITION MODES ...................................................................................................326
TOF MODE ...............................................................................................................................327
MS/MS MODES OF OPERATION................................................................................................328
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AUTO MS/MS...........................................................................................................................329
TARGETED MS/MS...................................................................................................................338
REFERENCE MASSES.................................................................................................................344
CHROMATOGRAM TAB..............................................................................................................347
PRE AND POST RUN SCRIPTS.....................................................................................................349
SAVE METHOD..........................................................................................................................351
RUN SINGLE INJECTION ............................................................................................................352
METHOD SAVED WITH DATA FILE ............................................................................................355
LAB EXERCISE: ACQUISITION ..........................................................................................357
IN THIS LABORATORY EXERCISE, YOU WILL NEED: ................................................................358
PREPARING THE LC/MSD Q-TOF ............................................................................................359
Q-TOF TOF ONLY ACQUISITION .............................................................................................360
AUTO MS/MS ACQUISITION.....................................................................................................363
TARGETED MS/MS ACQUISITION.............................................................................................365
WORKLISTS FOR AUTOMATION .......................................................................................367
IN THIS SECTION .......................................................................................................................368
WORKLIST PANE.......................................................................................................................369
CREATE ACQUISITION METHODS..............................................................................................370
NEW WORKLIST OR WORKLIST FROM TEMPLATE.....................................................................371
WORKLIST COLUMNS ...............................................................................................................372
ADD SAMPLES ..........................................................................................................................373
START WORKLIST .....................................................................................................................379
PARTIAL WORKLIST..................................................................................................................381
WAYS TO CREATE WORKLISTS .................................................................................................382
WORKLIST RUN PARAMETERS ..................................................................................................388
SCRIPTS ....................................................................................................................................391
WORKLIST REPORTS .................................................................................................................396
SAVE WORKLIST.......................................................................................................................399
LAB EXERCISE: WORKLISTS ..............................................................................................401
PREPARING THE LC/MSD Q-TOF ............................................................................................403
WORKLISTS...............................................................................................................................404