Book Content

1   History and Development of Liquid Chromatography   31

1.1   From the First Chromatographic Separations to High-Performance Liquid Chromatography   31

1.2   Development of High-Performance Liquid Chromatography in the New Millennium   37

 

2.1   Description of Retention in the Chromatographic System   41

2.2   Description of Selectivity in the Chromatographic System   45

2.3   Description of Efficiency in the Chromatographic System   46

2.4   Description of Resolution   70

2.5   Separations Using Gradient Elution   74

 

3.1   Basic Description of a Liquid Chromatograph   83

3.2   Mobile Phase Transport   85

3.3   Sample Injection   107

3.4   Chromatographic Columns for HPLC   119

3.5   Detection in HPLC   131

3.6   Derivatization in HPLC   182

 

4.1   Parameters Characterizing Stationary Phases   191

4.2   Silica Gel   198

4.3   Chemically Bonded Silica Gel Based Stationary Phases   205

4.4   Metal Oxide-Based Stationary Phases   234

4.5   Stationary Phases Based on Organic Polymers   240

4.6   Stationary Phases for Ion-Exchange Chromatography   242

4.7   Hybrid Stationary Phases   248

4.8   Porous Graphitic Carbon-Based Stationary Phases   261

4.9   Mixed-Mode Stationary Phases   264

 

5.1   Normal-Phase Chromatography (NP-HPLC)   279

5.2   Reversed-Phase Chromatography (RP-HPLC)   287

5.3   Ion-Exchange Chromatography (IEC)   296

5.4   Hydrophilic Interaction Liquid Chromatography (HILIC)   302

5.5   Hydrophobic Interaction Chromatography (HIC)   323

5.6   Mixed-Mode Chromatography   329

5.7   Ion-Pair Chromatography   335

5.8   Affinity Chromatography   341

5.9   Size-Exclusion Chromatography (SEC)   349

5.10   Chiral Chromatography   364

 

6.1   Ultra-High-Performance Liquid Chromatography (UHPLC)   400

6.2   Porous Shell Particles   418

6.3   Monolithic Columns   428

6.4   High-Temperature Liquid Chromatography (HTLC)   438

6.5   Supercritical Fluid Chromatography   446

6.6   Two-Dimensional Liquid Chromatography   459

6.7   Miniaturization in Liquid Chromatography   472

 

7.1   Mobile Phase Preparation and Use   481

7.2   Chromatographic Column – Care and Use   490

7.3   Regular Maintenance of HPLC System   504

 

8.1   Physicochemical Properties of the Analytes and Method Objective   507

8.2   Selection of Detection Approach   512

8.3   Selection of Chromatographic Mode   532

8.4   Selection of Stationary and Mobile Phases   542

8.5   Preliminary Experiments   549

8.6   Method Optimization   553

8.7   Solving Separation Problems in Method Development   564

8.8   Specific Optimization Approaches   566

 

9.1   Conventional Techniques for Sample Preparation   580

9.2   Modern Trends in Sample Preparation Techniques   612

 

10.1   Preparative HPLC   633

10.2   Preparative SFC   653

 

11.1   Qualitative Analysis   657

11.2   Quantitative Analysis   658

11.3   Metabolomics Approaches   677

 

12.1   Definitions of Metrological Terms   679

12.2   Validation Parameters   681

12.3   Practical Implementation of Validation   681

12.4   Pharmacopeial Methods   699

12.5   System Suitability Test   701

12.6   HPLC System Qualification   703

 

13.1   Back Pressure Issues   708

13.2   Baseline Issues   713

13.3   Retention Time Issues   722

13.4   Peak Shape and Peak Area Problems   724

13.5   Chromatogram Problems   739

 

 

15.1   Dynamic Viscosity of Acetonitrile/Water and Methanol/Water Mixtures   790

15.2   Miscibility of Organic Solvents Used in HPLC   791

15.3   Absorption Maxima and Molar Extinction Coefficients   792