Theory of HPLC

It is not the purpose of this chapter to go into the theory of HPLC, which is adequately covered in other texts, but to describe the applications of the method. This section will concentrate on analytical applications because semipreparative HPLC has been described in Section 1.3.2. Analytical HPLC finds use in the quantitative determination of plant constituents, in the purity control of natural products, and in chemotaxonomic investigations. 

This book describes the. hplc method and explains and illustrates its use. Each chapter deals with a different aspect of the method, beginning with an overview and ending with a detailed summary. Throughout, an attempt has been made to focus on questions related to the assay of the activity of an enzyme rather than its purification. More detailed discussions on the theory of hplc and on its use for purification, particularly for the purification of proteins, will be found in the references at the end of each chapter. 

J.H. Knox Theory of HPLC, Part II Solute Interactions with the Mobile Phase and Stationary Phases in Liquid Chromatography . In C.F. Simpson (ed.), Practical High Performance Liquid Chromatography. Heyden and Son, Chichester 1976. 

Although many have contributed to the theory of HPLC, only the summary by Kaizuma, H., Myers, M.N. and Giddings, J.C., J. of Chromatog. Sci., 8, 630 (1970) will be discussed now. Refer to the van Deemter section in Chapter 20, p. 217, to help you compare the theories of gas-liquid chromatography (GLC) and HPLC. In order to get the best separations, band broadening must be held to a minimum. The major factors that contribute to this are summarized in equation 19-1. 

Knox, J.H. (1976) Theory of HPLC, Part II solute interactions with the mobile phase and stationary phases in liquid chromatography, in Practical High Performance Liquid Chromatography (ed. C.F. Simpson), Heyden Son, Chichester. 

The characteristics of the different modes are treated in Chapters 2.1 to 2.7. It is assumed that the reader is familiar with the chromatographic rules and the theory of HPLC, so these are not treated in detail, but where necessary some technical terms are briefly explained. 

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