authors:subjects:- High performance liquid chromatography
publishers:ISBN:- 0471754013 (hbk.) :
- 9780471754015 (hbk.) :
description:notes:- Previous ed.: New York: VCH, 1994.
- Accompanied by CD-ROM in pocket.
- Includes bibliographical references and index.
- PREFACE. I HPLC PRIMER. 1 Advantages and Disadvantages of HPLC. 1.1 How It Works. 1.1.1 A Separation Model of the Column. 1.1.2 Basic Hardware: A Quick, First Look. 1.1.3 Use of Solvent Gradients. 1.1.4 Ranges of Compounds. 1.2 Other Ways to Make My Separation. 1.2.1 FPLC Fast Protein Liquid Chromatography. 1.2.2 LC Traditional Liquid Chromatography. 1.2.3 GLC Gas Liquid Chromatography. 1.2.4 SFC Supercritical Fluid Chromatography. 1.2.5 TLC Thin Layer Chromatography. 1.2.6 EP Electrophoresis. 1.2.7 CZE Capillary Zone Electrophoresis. 2 Selecting an HPLC System. 2.1 Characteristic Systems. 2.1.1 Finding a Fit: Detectors and Data Processing. 2.1.2 System Models: Gradient Versus Isocratic. 2.1.3 Vendor Selection. 2.1.4 Brand Names and Clones. 2.1.5 Hardware--Service--Support. 2.2 System Cost Estimates. 2.2.1 Type I System QC Isocratic (Cost: $10--15,000). 2.2.2 Type II System Research Gradient (Cost: $20--25,000). 2.2.3 Type III System Automated Clinical (Cost: $25--35,000). 2.2.4 Type IV System Automated Methods (Cost: $30--50,000). 2.3 Columns. 2.3.1 Sizes: Analytical and Preparative. 2.3.2 Separating Modes: Selecting Only What You Need. 2.3.3 Tips on Column Use. 3 Running Your Chromatograph. 3.1 Set--up and Start--up. 3.1.1 Hardware Plumbing 101: Tubing and Fittings. 3.1.2 Connecting Components. 3.1.3 Solvent Clean--up. 3.1.4 Water Purity Test. 3.1.5 Start--up System Flushing. 3.1.6 Column Preparation and Equilibration. 3.2 Sample Preparation and Column Calibration. 3.2.1 Sample Clean--up. 3.2.2 Plate Counts. 3.3 Your First Chromatogram. 3.3.1 Reproducible Injection Techniques. 3.3.2 Simple Scouting for a Mobile Phase. 3.3.3 Examining the Chromatogram. 3.3.4 Basic Calculations of Results. II HPLC OPTIMIZATION. 4 Separation Models. 4.1 Partition. 4.1.1 Separation Parameters. 4.1.2 Efficiency Factor. 4.1.3 Separation (Chemistry) Factor. 4.2 Ion Exchange Chromatography. 4.3 Size Exclusion Chromatography. 4.4 Affinity Chromatography. 5 Column Preparation. 5.1 Column Variations. 5.2 Packing Materials and Hardware. 5.3 Column Selection. 6 Column Aging, Diagnosis, and Healing. 6.1 Packing Degrading Bonded--Phase Loss. 6.2 Dissolved Packing Material End Voids. 6.3 Bound Material. 6.4 Pressure Increases. 6.5 Column Channeling Center--Voids. 6.6 Normal Phase, Ion Exchange, and Size Columns. 6.7 Zirconium and Polymer Columns. 7 Partition Chromatography Modifications. 7.1 Reverse--Phase and Hybrid Silica. 7.1.1 Ionization Suppression. 7.1.2 Ion Pairing. 7.1.3 Organic Modifiers. 7.1.4 Chelation. 7.2 Acidic Phase Silica. 7.3 Reverse--Phase Zirconium. 7.4 Partition Mode Selection. 8 "Nonpartition" Chromatography. 8.1 Ion Exchange. 8.1.1 Cationic:Weak and Strong. 8.1.2 Anionic:Weak and Strong. 8.2 Size Exclusion. 8.2.1 Organic Soluble Samples. 8.2.2 Hydrophilic Protein Separation. 8.3 Affinity Chromatography. 8.3.1 Column Packing Modification. 8.3.2 Chelation and Optically Active Columns. 9 Hardware Specifics. 9.1 System Protection. 9.1.1 Filters, Guard Columns, and Saturation Columns. 9.1.2 Inert Surfaces and Connections. 9.2 Pumping. 9.2.1 High-- and Low--Pressure Mixing Controllers. 9.2.2 Checking Gradient Performance. 9.3 Injectors and Autosamplers. 9.4 Detectors. 9.4.1 Mass Dependent Detectors. 9.4.2 Absorptive Detectors. 9.4.3 Specific Detectors. 9.5 Fraction Collectors. 9.6 Data Collection and Processing. 10 Troubleshooting and Optimization. 10.1 Hardware and Tools System Pacification. 10.2 Reverse Order Diagnosis. 10.3 Introduction to Data Acquisition. 10.4 Solvent Conservation. III HPLC UTILIZATION. 11 Preparative Chromatography. 11.1 Analytical Preparative. 11.2 Semipreparative. 11.3 "True" Preparative. 12 Sample Preparation and Methods Development. 12.1 Sample Preparation. 12.1.1 Deproteination. 12.1.2 Extraction and Concentration. 12.1.3 SFE (Cartridge Column) Preparations. 12.1.4 Extracting Encapsulated Compound
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