High Performance Liquid Chromatography, Softcover reprint of the original 1st ed. 1979
Chemical Laboratory Practice Series

Modern liquid column chromatography (LC) has developed rapidly since 1969 to become a standard method of separation. If the statisticians are to be believed, the recent growth of LC has been the most specta­ cular development in analytical chemistry and has not yet abated be­ cause its vast potential for application remains to be fully exploit­ ed. Significant factors contributing to this continued rise are the simplicity and low cost of the required basic equipment and the rela­ tive ease of acquiring and interpreting the data. Unfortunately, in LC, as so often in the field of analytical chemistry, the available commercial instruments are frequently far more complicated - and consequently far more expensive - than is nec­ essary for routine application. Therein also lies the risk of propa­ gating a "black box" philosophy that would be particularly detrimen­ tal to chromatography. Moreover, it appears to have been forgotten, as was done previously with gas chromatography, that inadequate sep­ aration by a column can be remedied only with great difficulty, if at all, by electronic means. Also, whether the capillary columns recent­ ly advocated with great enthusiasm for LC will fulfill the expecta­ tions of their proponents is highly questionable unless someone comes up with some new and revolutionary ideas.

I. Chromatographic Processes.- References Chapter I.- II. Fundamentals of Chromatography.- A. Retention.- B. Linear Flow Rate, Porosity, Permeability.- C. Band Broadening.- D. Resolution.- E. Dependence of Band Broadening on Flow Rate.- F. Band Broadening and Particle Size.- G. Extra-Column Band Broadening.- H. Optimum Analysis Conditions and Analysis Time.- I. Selection of a Suitable Column.- References Chapter II.- III. Equipment for HPLC.- A. Solvent Reservoir — Degassing of the Eluent.- B. Pumps.- 1. Syringe-Type Pumps.- 2. Reciprocating Piston Pumps and Diaphragm Pumps.- 3. Pumps with Variable Stroke Frequency.- 4. Gas-driven Displacement Pumps.- C. Damping of the Pulsations.- D. Sample Introduction.- 1. Sample Loops.- 2. Injection Devices.- E. The Column.- 1. Column Materials.- 2. Column Shape.- 3. Column Connections.- 4. Column Packing.- 5. Characterization and Testing of Columns.- F. Thermostating.- G. Measurement of the Flow Rate.- H. Fraction Collectors.- I. Recorders.- J. Instrumentation for Gradient Elution.- K. Safety Measures.- References Chapter III.- IV. Detectors.- A. UV Detectors.- B. Differential Refractometer.- 1. Fresnel-Refractometer.- 2. Deflection Refractometer.- C. Microadsorption Detector.- D. Transport Detector (Flame Ionization Detector).- E. Fluorescence Detector.- F. Other Detectors.- 1. Electrochemical Detectors.- 2. Conductivity Detector.- 3. Capacity Detector.- 4. Radioactivity Detectors.- 5. Directly Interfaced HPLC-Mass Spectrometry.- 6. Infra-red Detector.- 7. Other Methods.- G. Comparison of the Important Detectors.- H. Reaction Detectors.- References Chapter IV.- V. Stationary Phases.- A. Packing Materials for Adsorption and Partition Chromatography.- 1. Silica Gel.- 2. Alumina.- 3. Polyamides.- B. Chemically Modified Supports.- C. Ion Exchangers.- D. Stationary Phases for Exclusion Chromatography.- References Chapter V.- VI. Adsorption Chromatography.- I. Polar Stationary Phases.- A. General.- B. Stationary Phases.- C. Effect of Water on Separations.- D. Effect of Eluent on Separation.- 1. Eluotropic Series.- 2. Choice of Eluent.- 3. Solvent Mixtures.- E. Effect of Sample Structure.- II. Nonpolar Stationary Phases.- A. General.- B. Reversed Phase Properties.- C. Effect of Solvent on Separation.- D. Effect of Sample Structure.- III. The General Elution Problem.- A. Pressure or Flow Programming.- B. Temperature Programming.- C. Programming of the Stationary Phase.- 1. Variation of Adsorbent Activity.- 2. Coupled Columns.- D. Gradient Elution. Programming of the Eluent Composition.- IV. Applications of Adsorption Chromatography.- A. On Polar Stationary Phases.- B. On Nonpolar Stationary Phases.- C. Separations on Polyamide.- References Chapter VI.- VII. Partition Chromatography.- A. Introduction.- B. Supports and Liquid Phases.- 1. Supports.- 2. Liquid Phases.- 3. Coating the Support.- 4. Determination of the Coverage.- C. Properties of the Column.- 1. Stability.- 2. Sample Capacity.- 3. Preparative Applications.- 4. Column Efficiency.- 5. Programming Techniques.- D. Applications.- E. Ion-pair Chromatography.- References Chapter VII.- VIII. Ion-Exchange Chromatography.- A. Principle.- B. Ion Exchange Materials.- 1. Ion Exchangers with an Organic Polymer Matrix.- 2. Polymeric Ion Exchangers on PLB Particles.- 3. Brush Type Ion Exchangers.- 4. Liquid Ion Exchangers.- C. Characterization of the Ion Exchangers.- D. Optimizing a Separation.- 1. The Effect of pH on Retention.- 2. Effect of Ionic Strength on Retention.- 3. Change of the Buffer Solution.- 4. Other Effects.- 5. Gradient Elution.- E. Applications.- 1. Classical Ion Exchangers in HPLC.- 2. Porous Layer Beads (PLB).- 3. Ion Exchangers on Chemically Modified Silica Gel.- References Chapter VIII.- IX. Exclusion Chromatography. Gel Permeation Chromatography.- A. Introduction.- B. Basics of Exclusion Chromatography.- C. Stationary Phases for Exclusion Chromatography.- D. Applications of Exclusion Chromatography.- 1. Determination of the Molecular Weight Distribution of Polymers.- 2. Application of Rapid Exclusion Chromatography to Biochemical Problems.- References Chapter IX.- X. Selection of the Separation System.- References Chapter X.- XI. Special Techniques.- A. Preparative Chromatography.- B. Qualitative Analysis.- C. Quantitative Analysis.- D. Trace Analysis.- References Chapter XI.- XII. Purification of Solvents.- References Chapter XII.