HPLC

Name: HPLC | A Practical User's Guide
Brand: Wiley
Price: 96.99 EUR
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A Practical User's Guide

Marvin McMaster(Autor*in)

Wiley (Verlag)

2. Auflage

Erschienen am 4. Januar 2007

256 Seiten

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978-0-470-07908-9 (ISBN)

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Inhalt

Intro
HPLC
CONTENTS
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 Compounds
12.1.5 SFE Trace Enrichment and Windowing
12.1.6 Derivatives
12.2 Methods Development
12.2.1 Standards Development
12.2.2 Samples Development
12.3 Gradient Development
13 Application Logics: Separations Overview
13.1 Fat-Soluble Vitamins, Steroid, and Lipids
13.2 Water-Soluble Vitamins, Carbohydrates, and Acids
13.3 Nucleomics
13.4 Proteomics
13.5 Clinical and Forensic Drug Monitoring
13.6 Pharmaceutical Drug Development
13.7 Environmental and Reaction Monitoring
13.8 Application Trends
14 Automation
14.1 Analog-to-Digital Interfacing
14.2 Digital Information Exchange
14.3 HPLC System Control and Automation
14.4 Data Collection and Interpretation
14.4.1 Preinjection Baseline Setting
14.4.2 Peak Detection and Integration
14.4.3 Quantitation: Internal/External Standards
14.5 Automated Methods Development
14.5.1 Automated Isocratic Development
14.5.2 Hinge Point Gradient Development
14.6 Data Exportation to the Real World
14.6.1 Word Processors: .ASC, .DOC, .RTF, .WS, .WP Formats
14.6.2 Spread Sheets: .DIF, .WK, .XLS Formats
14.6.3 Databases: .DB2 Format
14.6.4 Graphics: .PCX, .TIFF, .JPG Formats
14.6.5 Chromatographic Files: Metafiles and NetCDF
15 Recent Advances in LC/MS Separations
15.1 A LC/MS Primer
15.1.1 Quadrupole MS and Mass Selection
15.1.2 Other Types of MS Analyzers for LC/MS
15.1.3 LC/MS Interfaces
15.1.4 LC/MS Computer Control and Data Processing
15.2 Microflow Chromatography
15.3 Ultrafast HPLC Systems
15.4 Chip HPLC Systems
15.5 Standardized LC/MS in Drug Design
16 New Directions in HPLC
16.1 Temperature-Controlled Chromatography
16.2 Ultrafast Chromatography
16.3 Monolith Capillary Columns
16.4 Micro-Parallel HPLC Systems
16.5 Two-Dimensional HPLC Systems
16.6 The Portable LC/MS
APPENDICES
APPENDIX A Personal Separations Guide
APPENDIX B FAQs for HPLC Systems and Columns
APPENDIX C Tables of Solvents and Volatile Buffers
APPENDIX D Glossary of HPLC Terms
APPENDIX E HPLC Troubleshooting Quick Reference
APPENDIX F HPLC Laboratory Experiments
Laboratory 1-System Start-up and Column Quality Control
Laboratory 2-Sample Preparation and Methods Development
Laboratory 3-Column and Solvent Switching and Pacification
Appendix G Selected Reference List
INDEX

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