Since the first edition of this book the major advances have been in column packings, where over ninety per cent of separations are now performed using chemically bonded microparticulate packings, and in instrumentation. The use of microprocessor control has brought about a rationalization of mobile phase delivery systems and in detectors, the introduction of electrochemical and spectrophotometric detection other than in the ultra-violet region, has widened the field of applications and the sensitivity of the technique. The use of ion­ pair chromatography has increased at the expense of ion-exchange and this together with the improvements in detectors has greatly increased the appli­ cation of the technique in the biomedical field. These advances are described together with the established methods to enable the beginner to carry out a satisfactory separation and to gain the experience necessary for the full exploitation of the technique. R. J. Hamilton P. A. Sewell Liverpool,1981 1 Introduction to high performance liquid chromatography 1. 1 Introduction Chromatography in its many forms is widely used as a separative and an analytical technique. Gas chromatography since its introduction by James and Martin [1] has been pre-eminent in the field. Uquid chromatography in the of paper, thin-layer, ion-exchange, and exclusion (gel permeation and gel form filtration) chromatography had not been able to achieve the same success, mainly because of the poor efficiences and the long analysis times arising from the low mobile phase flow rates.

Inhalt

1 Introduction to high performance liquid chromatography.- 1.1 Introduction.- 1.2 Nomenclature.- 1.3 Liquid Chromatography Modes.- 1.4 Scope of Techniques.- References.- 2 Chromatographic theory.- 2.1 The Process of Separation.- 2.2 Retention in Liquid Chromatography.- 2.3 Band Broadening-Origins.- 2.4 Band Broadening and the Plate Height Equation.- 2.5 Overall Plate Height Equation.- 2.6 Comparison with Gas Chromatography.- 2.7 Column Efficiency and Particle Diameter.- 2.8 Reduced Plate Height and Reduced Velocity.- 2.9 Extra-Column Band Broadening.- 2.10 Resolution.- 2.11 Resolution and Time for Analysis.- 2.12 Theory of Exclusion Chromatography.- References.- 3 Equipment.- 3.1 Introduction.- 3.2 Mobile Phase (Solvent) Reservoirs and Solvent Degassing.- 3.3 Pumping Systems.- 3.4 Flow Controllers.- 3.5 Solvent Flow Programming Equipment.- 3.6 Pulse Damping.- 3.7 Pressure Measurement.- 3.8 Filters.- 3.9 Sample Introduction Systems.- 3.10 Columns and Column Fittings.- 3.11 Column Thermostats.- 3.12 Liquid Chromatography Detectors.- 3.13 Flow rate Measurement.- 3.14 Fraction Collectors.- 3.15 Data Handling.- 3.16 Microprocessor Controlled HPLC.- References.- 4. Stationary phases in liquid chromatography.- 4.1 Introduction.- 4.2 Stationary Phase Types.- 4.3 Coltimn Packing Techniques.- 4.4 Liquid-Solid Chromatography.- 4.5 Liquid-Liquid Chromatography.- 4.6 Ion-Exchange Chromatography.- 4.7 Exclusion Chromatography.- 4.8 Care of Columns.- References.- 5. Mobile phases in liquid chromatography.- 5.1 Introduction.- 5.2 Solvent Qualities.- 5.3 liquid-Solid Chromatography.- 5.4 Liquid-Liquid Chromatography.- 5.5 Ion-Exchange Chromatography.- 5.6 Exclusion Chromatography.- 5.7 Gradient Elution.- References.- 6. Developing a chromatogram.- 6.1 Nature of the Problem.- 6.2 Choice of Chromatographic Mode.- 6.3 Selection of Stationary Phase and Mobile Phase.- 6.4 Choice of Detector.- 6.5 Chromatographic Separation.- 6.6 The General Elution Problem.- 6.7 Quantitative Analysis.- References.- 7. Preparative high performance liquid chromatography and trace analysis.- 7.1 Introduction.- 7.2 Stages in Preparative HPLC.- 7.3 Infinite Diameter Columns.- 7.4 Packing of a Preparative Column.- 7.5 Summary of Preparative HPLC.- 7.6 Trace Analysis.- References.- 8. Applications of high performance liquid chromatography.- 8.1 Pharmaceuticals.- 8.2 Biochemicals.- 8.3 Food Chemicals.- 8.4 Heavy Industrial Chemicals.- 8.5 Inorganic.- 8.6 Miscellaneous.- Compound Index.