LC/MS：A Pratical User's Guide

Preface

1 Introduction to LC/MS 1
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 9
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 21
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
10 Application Areas for LC/MS 95
10.1 Compound Discovery, 95
10.2 Identification of Complex Biological Compounds, 96
10.3 Analysis of Trace Impurities and Metabolites, 97
10.4 Arson Residue Investigation, 98
10.5 Industrial Water and Pesticide Analysis, 98
10.6 Toxicology and Drugs of Abuse, 98
10.7 Clinical Therapeutic Drug Screening, 99
10.8 Pesticide Manufacturing, 101
11 Trace Analysis and LC/MS/MS 103
11.1 LC/MS/MS Triple-Quadrupole System, 103
11.2 MS/MS Operating Modes, 104
11.3 Ion Trap MS/MS Operation, 106
11.4 Hybrid LC/MS/MS Systems, 108
12 Drug Discovery and Benchtop LC/MS 111
12.1 Activity Screening, 111
12.2 Standardized LC/MS Screening, 113
12.3 Molecular Fragmentation for Structural Determination, 115
12.4 Process Monitoring, 116
13 Proteomics: LC/MALDI/TOF and MS/MS Libraries 119
13.1 Protein Molecular-Weight Determination by LC/MS, 120
13.2 De Novo Protein Purification, 121
13.3 Protein Analysis by Two-Dimensional GEP and
LC/TOFMS, 122
13.4 LC/MS/MS Identification of Peptide Structures, 122
13.5 Tracer Labeling for Peptide ID, 124
13.6 Posttranslational Modified Protein, 124
13.7 Transient Peptides and Accumulation Proteins, 124
14 The Future of LC/MS 127
14.1 Instrumentation Improvements, 127
14.2 Affordable Benchtop LC/LITMS, 129
14.3 User-Customized Data Libraries, 129
14.4 Nucleomics and Restriction Fragment Analysis, 130
Appendix A LC/MS Frequently Asked Questions 131
Appendix B Solvents and Volatile Buffers for LC/MS 139
Appendix C Guide to Structure Interpretation 143
Appendix D Glossary of LC/MS Terms 149
Appendix E LC/MS Selective Reading List 155
Index 157