1 Introduction to LC/MS
1.1 Why LC/MS?................................................. 1
1.2 Molecular Weights and Structure Studies ................... 4
1.3 LC/MS Systems ............................................. 4
1.4 System Costs .............................................. 7
1.5 Competitive Systems ....................................... 7
2 The HPLC System
2.1 HPLC System Components .................................... 9
2.2 Gradient versus Isocratic Systems ........................ 14
2.3 Micro HPLC Systems ....................................... 16
2.4 HPLC Tubing and Fittings ................................. 18
3 The HPLC Column and Separation Modes
3.1 Column Construction ...................................... 21
3.2 Column Packing Materials ................................. 23
3.3 Normal-Phase Columns ..................................... 25
3.4 Other Bonded-Phase Silica Columns ........................ 26
3.5 Optimizing Reverse-Phase Column Use ...................... 28
3.6 Silica Ion-Exchange Columns .............................. 30
3.7 Silica Size-Separation Columns ........................... 31
3.8 Zirconium Bonded-Phase Columns ........................... 31
3.9 Polymer Reverse-Phase Columns ............................ 32
4 HPLC and Column Maintenance .................................. 33
4.1 HPLC Maintenance ......................................... 33
4.2 Column Maintenance ....................................... 37
5 Sample Preparation and Separations Development ............... 41
5.1 Mobile-Phase Preparation ................................. 41
5.2 Mobile-Phase pH Control Using Buffers .................... 42
5.3 Sample Preparation ....................................... 44
5.4 Cartridge Column Cleanup ................................. 44
5.5 On-Column Sample Concentration ........................... 45
5.6 Isocratic and Gradient Methods Development ............... 46
5.7 Automated Methods Development ............................ 49
6 LC/MS Interfaces ............................................. 51
6.1 Solvent Removal and Ionization ........................... 51
6.2 Atmospheric-Pressure Interfaces .......................... 52
6.3 Electrospray Interface .................................. 53
6.4 Ion Spray Interface ...................................... 54
6.5 Secondary Detectors ...................................... 55
7 LC/MS Overview ............................................... 59
7.1 HPLC and the Ionization Source .......................... 60
7.2 Vacuum Pumps ............................................. 61
7.3 Analyzer and Ion Detector Designs ........................ 61
7.4 Data and Control Systems ................................. 66
7.5 Peak Detection, ID, and Quantitation .................... 69
8 Mass Analyzers ............................................... 71
8.1 Quadrupole Analyzer ...................................... 71
8.2 Ion Trap Analyzer ........................................ 74
8.3 Linear Ion Trap Analyzer ................................. 78
8.4 Time-of-Flight Analyzer .................................. 79
8.5 Fourier Transform Analyzer Design ........................ 81
8.6 Magnetic Sector Analyzers ................................ 83
9 Mass Spectrometer Maintenance ................................ 85
9.1 High-Vacuum Operation .................................... 85
9.2 MS Hardware Maintenance ................................. 88
9.3 System Electrical Grounding .............................. 92
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