HPLC: A Practical User's Guide

摘要: PREFACE. I HPLC PRIMER. 1 Advantages and Disadvantages of HPLC. 1.1 How It Works. 1.1.1 A Separation Model the Column. 1.1.2 Basic Hardware: Quick, First Look. 1.1.3 Use Solvent Gradients. 1.1.4 Ranges Compounds. 1.2 Other Ways to Make My Separation. 1.2.1 FPLC-Fast Protein Liquid Chromatography. 1.2.2 LC-Traditional 1.2.3 GLC-Gas 1.2.4 SFC-Supercritical Fluid 1.2.5 TLC-Thin Layer 1.2.6 EP-Electrophoresis. 1.2.7 CZE-Capillary Zone Electrophoresis. 2 Selecting an System. 2.1 Characteristic Systems. 2.1.1 Finding a Fit: Detectors Data Processing. 2.1.2 System Models: Gradient Versus Isocratic. 2.1.3 Vendor Selection. 2.1.4 Brand Names Clones. 2.1.5 Hardware-Service-Support. 2.2 Cost Estimates. 2.2.1 Type System-QC Isocratic (Cost: $10-15,000). 2.2.2 II System-Research $20-25,000). 2.2.3 III System-Automated Clinical $25-35,000). 2.2.4 IV Methods $30-50,000). 2.3 Columns. 2.3.1 Sizes: Analytical Preparative. 2.3.2 Separating Modes: Only What You Need. 2.3.3 Tips on Column Use. 3 Running Your Chromatograph. 3.1 Set-up Start-up. 3.1.1 Hardware Plumbing 101: Tubing Fittings. 3.1.2 Connecting Components. 3.1.3 Clean-up. 3.1.4 Water Purity Test. 3.1.5 Start-up Flushing. 3.1.6 Preparation Equilibration. 3.2 Sample Calibration. 3.2.1 3.2.2 Plate Counts. 3.3 Chromatogram. 3.3.1 Reproducible Injection Techniques. 3.3.2 Simple Scouting for Mobile Phase. 3.3.3 Examining 3.3.4 Calculations Results. OPTIMIZATION. 4 Models. 4.1 Partition. 4.1.1 Parameters. 4.1.2 Efficiency Factor. 4.1.3 (Chemistry) 4.2 Ion Exchange 4.3 Size Exclusion 4.4 Affinity 5 Preparation. 5.1 Variations. 5.2 Packing Materials Hardware. 5.3 6 Aging, Diagnosis, Healing. 6.1 Degrading-Bonded-Phase Loss. 6.2 Dissolved Material-End Voids. 6.3 Bound Material. 6.4 Pressure Increases. 6.5 Channeling-Center-Voids. 6.6 Normal Phase, Exchange, 6.7 Zirconium Polymer 7 Partition Chromatography Modifications. 7.1 Reverse-Phase Hybrid Silica. 7.1.1 Ionization Suppression. 7.1.2 Pairing. 7.1.3 Organic Modifiers. 7.1.4 Chelation. 7.2 Acidic Phase 7.3 Zirconium. 7.4 Mode 8 "Nonpartition" 8.1 Exchange. 8.1.1 Cationic:Weak Strong. 8.1.2 Anionic:Weak 8.2 Exclusion. 8.2.1 Soluble Samples. 8.2.2 Hydrophilic 8.3 8.3.1 Modification. 8.3.2 Chelation Optically Active 9 Specifics. 9.1 Protection. 9.1.1 Filters, Guard Columns, Saturation 9.1.2 Inert Surfaces Connections. 9.2 Pumping. 9.2.1 High- Low-Pressure Mixing Controllers. 9.2.2 Checking Performance. 9.3 Injectors Autosamplers. 9.4 Detectors. 9.4.1 Mass Dependent 9.4.2 Absorptive 9.4.3 Specific 9.5 Fraction Collectors. 9.6 Collection 10 Troubleshooting Optimization. 10.1 Tools-System Pacification. 10.2 Reverse Order Diagnosis. 10.3 Introduction Acquisition. 10.4 Conservation. UTILIZATION. 11 Preparative 11.1 11.2 Semipreparative. 11.3 "True" 12 Development. 12.1 12.1.1 Deproteination. 12.1.2 Extraction Concentration. 12.1.3 SFE (Cartridge Column) Preparations. 12.1.4 Extracting Encapsulated 12.1.5 Trace Enrichment Windowing. 12.1.6 Derivatives. 12.2 12.2.1 Standards 12.2.2 Samples 12.3 13 Application Logics: Separations Overview. 13.1 Fat-Soluble Vitamins, Steroid, Lipids. 13.2 Water-Soluble Carbohydrates, Acids. 13.3 Nucleomics. 13.4 Proteomics. 13.5 Forensic Drug Monitoring. 13.6 Pharmaceutical 13.7 Environmental Reaction 13.8 Trends. 14 Automation. 14.1 Analog-to-Digital Interfacing. 14.2 Digital Information 14.3 Control 14.4 Interpretation. 14.4.1 Preinjection Baseline Setting. 14.4.2 Peak Detection Integration. 14.4.3 Quantitation: Internal/External Standards. 14.5 Automated 14.5.1 14.5.2 Hinge Point 14.6 Exportation Real World. 14.6.1 Word Processors: .ASC, .DOC, .RTF, .WS, .WP Formats. 14.6.2 Spread Sheets: .DIF, .WK, .XLS 14.6.3 Databases: .DB2 Format. 14.6.4 Graphics: .PCX, .TIFF, .JPG 14.6.5 Chromatographic Files: Metafiles NetCDF. 15 Recent Advances in LC/MS Separations. 15.1 Primer. 15.1.1 Quadrupole MS 15.1.2 Types Analyzers LC/MS. 15.1.3 Interfaces. 15.1.4 Computer 15.2 Microflow 15.3 Ultrafast 15.4 Chip 15.5 Standardized Design. 16 New Directions 16.1 Temperature-Controlled 16.2 16.3 Monolith Capillary 16.4 Micro-Parallel 16.5 Two-Dimensional 16.6 The Portable APPENDICES. APPENDIX Personal Guide. B FAQs Systems C Tables Solvents Volatile Buffers. D Glossary Terms. E Quick Reference. F Laboratory Experiments. 1: Quality Control. 2: 3: Switching Appendix G Selected Reference List. INDEX.