Table of Contents
Cover
Title
Copyright
Dedication
Preface to the Fifth Edition
Important and Useful Equations for HPLC
1 Introduction
1.1 HPLC: A powerful separation method
1.2 A first HPLC experiment
1.3 Liquid chromatographic separation modes
1.4 The HPLC instrument
1.5 Safety in the HPLC laboratory
1.6 Comparison between high-performance liquid chromatography and gas chromatography
1.7 Comparison between high-performance liquid chromatography and capillary electrophoresis
1.8 Units for pressure, length and viscosity
1.9 Scientific journals
1.10 Recommended books
2 Theoretical Principles
2.1 The chromatographic process
2.2 Band broadening
2.3 The chromatogram and its purport
2.4 Graphical representation of peak pairs with different degree of resolution
2.5 Factors affecting resolution
2.6 Extra-column volumes (dead volumes)
2.7 Tailing
2.8 Peak capacity and statistical resolution probability
2.9 Effects of temperature in HPLC
2.10 The limits of HPLC
2.11 How to obtain peak capacity
3 Pumps
3.1 General requirements
3.2 The short-stroke piston pump
3.3 Maintenance and repair
3.4 Other pump designs
4 Preparation of Equipment up to Sample Injection
4.1 Selection of the mobile phase
4.2 Preparation of the mobile phase
4.3 Gradient systems
4.4 Capillary tubing
4.5 Fittings
4.6 Sample injectors
4.7 Sample solution and sample volume
5 Solvent Properties
5.1 Table of organic solvents
5.2 Solvent selectivity
5.3 Miscibility
5.4 Buffers
5.5 Shelf life of mobile phases
5.6 The mixing cross
6 Detectors
6.1 General
6.2 UV detectors
6.3 Refractive index detectors
6.4 Fluorescence detectors
6.5 Electrochemical (amperometric) detectors
6.6 Light-scattering detectors
6.7 Other detectors
6.8 Multiple detection
6.9 Indirect detection
6.10 Coupling with spectroscopy
7 Columns and Stationary Phases
7.1 Columns for HPLC
7.2 Precolumns
7.3 General properties of stationary phases
7.4 Silica
7.5 Chemically modified silica
7.6 Styrene-divinylbenzene
7.7 Some other stationary phases
7.8 Column care and regeneration
8 HPLC Column Tests
8.1 Simple tests for HPLC columns
8.2 Determination of particle size
8.3 Determination of breakthrough time
8.4 The test mixture
8.5 Dimensionless parameters for HPLC column characterization
8.6 The van Deemter equation from reduced parameters and its use in column diagnosis
8.7 van Deemter curves and other coherences
8.8 Diffusion coefficients
9 Adsorption Chromatography: Normal-Phase Chromatography
9.1 What is adsorption?
9.2 The eluotropic series
9.3 Selectivity properties of the mobile phase
9.4 Choice and optimization of the mobile phase
9.5 Applications
10 Reversed-Phase Chromatography
10.1 Principle
10.2 Mobile phases in reversed-phase chromatography
10.3 Solvent selectivity and strength
10.4 Stationary phases
10.5 Method development in reversed-phase chromatography
10.6 Applications
10.7 Hydrophobic interaction chromatography
11 Chromatography with Chemically Bonded Phases
11.1 Introduction
11.2 Properties of some stationary phases
11.3 Hydrophilic interaction chromatography
12 Ion-Exchange Chromatography
12.1 Introduction
12.2 Principle
12.3 Properties of ion exchangers
12.4 Influence of the mobile phase
12.5 Special possibilities of ion exchange
12.6 Practical hints
12.7 Applications
13 Ion-Pair Chromatography
13.1 Introduction
13.2 Ion-pair chromatography in practice
13.3 Applications
13.4 Appendix: UV detection using ion-pair reagents
14 Ion Chromatography
14.1 Principle
14.2 Suppression techniques
14.3 Phase systems
14.4 Applications
15 Size-Exclusion Chromatography
15.1 Principle
15.2 The calibration chromatogram
15.3 Molecular mass determination by means of size-exclusion chromatography
15.4 Coupled size-exclusion columns
15.5 Phase systems
15.6 Applications
16 Affinity Chromatography
16.1 Principle
16.2 Affinity chromatography as a special case of HPLC
16.3 Applications
17 Choice of Method
17.1 The various possibilities
17.2 Method transfer
18 Solving the Elution Problem
18.1 The elution problem
18.2 Solvent gradients
18.3 Column switching
18.4 Comprehensive two-dimensional HPLC
18.5 Optimization of an isocratic chromatogram using four solvents
18.6 Optimization of the other parameters
18.7 Mixed stationary phases
19 Analytical HPLC
19.1 Qualitative analysis
19.2 Trace analysis
19.3 Quantitative analysis
19.4 Recovery
19.5 Peak-height and peak-area determination for quantitative analysis
19.6 Integration errors
19.7 The detection wavelength
19.8 Derivatization
19.9 Unexpected peaks: Ghost and system peaks
20 Quality Assurance
20.1 Is it worth the effort?
20.2 Verification with a second method
20.3 Method validation
20.4 Standard operating procedures
20.5 Measurement uncertainty
20.6 Qualifications, instrument test and system suitability test
20.7 The quest for quality
21 Preparative HPLC
21.1 Problem
21.2 Preparative HPLC in practice
21.3 Overloading effects
21.4 Fraction collection
21.5 Recycling
21.6 Displacement chromatography
22 Separation of Enantiomers
22.1 Introduction
22.2 Chiral mobile phases
22.3 Chiral liquid stationary phases
22.4 Chiral solid stationary phases
22.5 Indirect separation of enantiomers
23 Special Possibilities
23.1 Micro, capillary and chip HPLC
23.2 High-speed and super-speed HPLC
23.3 Fast separations at 1000 bar: UHPLC
23.4 HPLC with supercritical mobile phases
23.5 HPLC with superheated water
23.6 Electrochromatography
24 Appendix 1: Applied HPLC Theory
25 Appendix 2: How to Perform the Instrument Test
25.1 Introduction
25.2 Test sequence
25.3 Preparations
25.4 Pump test
25.5 UV detector test
25.6 Autosampler test
25.7 Column oven test
25.8 Equations and calculations
25.9 Documentation
26 Appendix 3: Troubleshooting
26.1 Pressure problems
26.2 Leak in the pump system
26.3 Deviating retention times
26.4 Injection problems
26.5 Baseline problems
26.6 Peak shape problems
26.7 Problems with light-scattering detectors
26.8 Other causes
26.9 Instrument test
27 Appendix 4: Column Packing
Index of Separations
Subject Index
End User License Agreement
Guide
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