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[资源] HPLC：A Practical User's Guide[2007]

Table of Contents
Preface    xi
HPLC Primer    1
Advantages and Disadvantages of HPLC    3
How It Works    4
A Separation Model of the Column    5
Basic Hardware: A Quick, First Look    7
Use of Solvent Gradients    8
Ranges of Compounds    9
Other Ways to Make My Separation    9
FPLC-Fast Protein Liquid Chromatography    10
LC-Traditional Liquid Chromatography    10
GLC-Gas Liquid Chromatography    11
SFC-Supercritical Fluid Chromatography    11
TLC-Thin Layer Chromatography    12
EP-Electrophoresis    12
CZE-Capillary Zone Electrophoresis    13
Selecting an HPLC System    15
Characteristic Systems    16
Finding a Fit: Detectors and Data Processing    16
System Models: Gradient Versus Isocratic    16
Vendor Selection    17
Brand Names and Clones    17
Hardware-Service-Support    18
System Cost Estimates    19
Type 1 System-QC Isocratic (Cost: {dollar}10-15,000)    19
Type II System-Research Gradient (Cost: {dollar}20-25,000)    19
Type III System-Automated Clinical(Cost: {dollar}25-35,000)    20
Type IV System-Automated Methods (Cost: {dollar}30-50,000)    21
Columns    21
Sizes: Analytical and Preparative    21
Separating Modes: Selecting Only What You Need    22
Tips on Column Use    23
Running Your Chromatograph    25
Set-up and Start-up    25
Hardware Plumbing 101: Tubing and Fittings    26
Connecting Components    28
Solvent Clean-up    30
Water Purity Test    33
Start-up System Flushing    34
Column Preparation and Equilibration    35
Sample Preparation and Column Calibration    36
Sample Clean-up    36
Plate Counts    37
Your First Chromatogram    37
Reproducible Injection Techniques    38
Simple Scouting for a Mobile Phase    39
Examining the Chromatogram    40
Basic Calculations of Results    41
HPLC Optimization    43
Separation Models    45
Partition    45
Separation Parameters    48
Efficiency Factor    49
Separation (Chemistry) Factor    53
Ion Exchange Chromatography    56
Size Exclusion Chromatography    57
Affinity Chromatography    59
Column Preparation    61
Column Variations    61
Packing Materials and Hardware    64
Column Selection    66
Column Aging, Diagnosis, and Healing    73
Packing Degrading-Bonded-Phase Loss    74
Dissolved Packing Material-End Voids    77
Bound Material    78
Pressure Increases    81
Column Channeling-Center-Voids    83
Normal Phase, Ion Exchange, and Size Columns    84
Zirconium and Polymer Columns    86
Partition Chromatography Modifications    89
Reverse-Phase and Hybrid Silica    89
Ionization Suppression    90
Ion Pairing    91
Organic Modifiers    92
Chelation    92
Acidic Phase Silica    93
Reverse-Phase Zirconium    93
Partition Mode Selection    94
"Nonpartition" Chromatography    95
Ion Exchange    96
Cationic: Weak and Strong    96
Anionic: Weak and Strong    97
Size Exclusion    98
Organic Soluble Samples    98
Hydrophilic Protein Separation    99
Affinity Chromatography    101
Column Packing Modification    102
Chelation and Optically Active Columns    103
Hardware Specifics    105
System Protection    105
Filters, Guard Columns, and Saturation Columns    106
Inert Surfaces and Connections    107
Pumping    108
High-and Low-Pressure Mixing Controllers    109
Checking Gradient Performance    112
Injectors and Autosamplers    113
Detectors    116
Mass Dependent Detectors    116
Absorptive Detectors    119
Specific Detectors    122
Fraction Collectors    123
Data Collection and Processing    123
Troubleshooting and Optimization    125
Hardware and Tools-System Pacification    125
Reverse Order Diagnosis    129
Introduction to Data Acquisition    132
Solvent Conservation    133
HPLC Utilization    135
Preparative Chromatography    137
Analytical Preparative    138
Semipreparative    139
"True" Preparative    139
Sample Preparation and Methods Development    143
Sample Preparation    143
Deproteination    144
Extraction and Concentration    145
SFE (Cartridge Column) Preparations    145
Extracting Encapsulated Compounds    147
SFE Trace Enrichment and Windowing    148
Derivatives    151
Methods Development    151
Standards Development    152
Samples Development    154
Gradient Development    156
Application Logics: Separations Overview    159
Fat-Soluble Vitamins, Steroid, and Lipids    159
Water-Soluble Vitamins, Carbohydrates, and Acids    160
Nucleomics    161
Proteomics    162
Clinical and Forensic Drug Monitoring    163
Pharmaceutical Drug Development    164
Environmental and Reaction Monitoring    164
Application Trends    165
Automation    167
Analog-to-Digital Interfacing    167
Digital Information Exchange    169
HPLC System Control and Automation    169
Data Collection and Interpretation    170
Preinjection Baseline Setting    171
Peak Detection and Integration    171
Quantitation: Internal/External Standards    172
Automated Methods Development    172
Automated Isocratic Development    173
Hinge Point Gradient Development    176
Data Exportation to the Real World    177
Word Processors; .ASC, .DOC, .RTF, .WS, .WP Formats    177
Spread Sheets: .DIF, .WK, .XLS Formats    178
Databases: .DB2 Format    178
Graphics: .PCX, .TIFF, .JPG Formats    178
Chromatographic Files: Metafiles and NetCDF    178
Recent Advances in LC/MS Separations    181
A LC/MS Primer    181
Quadrupole MS and Mass Selection    183
Other Types of MS Analyzers for LC/MS    185
LC/MS Interfaces    187
LC/MS Computer Control and Data Processing    189
Microflow Chromatography    191
Ultrafast HPLC Systems    192
Chip HPLC Systems    192
Standardized LC/MS in Drug Design    193
New Directions in HPLC    195
Temperature-Controlled Chromatography    195
Ultrafast Chromatography    196
Monolith Capillary Columns    196
Micro-Parallel HPLC Systems    197
Two-Dimensional HPLC Systems    197
The Portable LC/MS    198
Appendices    201
Personal Separations Guide    203
FAQs for HPLC Systems and Columns    205
Tables of Solvents and Volatile Buffers    211
Glossary of HPLC Terms    213
HPLC Troubleshooting Quick Reference    221
HPLC Laboratory Experiments    227
System Start-up and Column Quality Control    227
Sample Preparation and Methods Development    229
Column and Solvent Switching and Pacification    231
Selected Reference List    233
Index    235

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