Description

The Practical Approach series of books aims to provide sufficient information on an analytical technique or area of study to enable readers to begin work in that field. This particular book is the second edition of a book first published in 1989. The advances in HPLC instrumentation in the intervening ten years have necessitated a substantial re-write of the book with the addition of four new chapters.

The book contains seven chapters: chapters 1 and 2 are devoted to the developments in microbore HPLC and the introduction of mass spectrometric detectors for liquid chromatography, chapters 3, 4 and 5 describe methods for the separation of proteins and peptides by size-exclusion, ion-exchange and reversed-phase chromatography respectively, while chapters 6 and 7 reflect the increasing interest in HPLC separation of polynucleotides and glycoproteins.

Most chromatographers will be used to the standard 4.6 mm internal diameter HPLC columns, but the use of narrower, down to 1 mm i.d., columns has many theoretical advantages. Chapter 1 describes the changes to standard HPLC instrumentation required to operate in microbore mode, including the need for enhanced detector capabilities. This is complemented by an excellent chapter on the advances in sample introduction techniques which have permitted the connection of mass spectrometers to liquid chromatography systems without risk of drowning. The diagrams in this chapter are particularly clear and help the reader understand the principles behind the various sampling systems now available.

The remaining chapters follow the standard principles of the Practical Approach series in that they contain an introduction to the fundamentals of the particular technique, details of instrumentation or consumables required and sample protocols for separations to assist the analyst new to the technique. As this particular book is aimed at a general biochemistry audience, few of the examples are of direct interest to the clinical biochemist, concentrating on separations of proteins or peptides in standard mixtures. The chapter on reversed-phase chromatography of proteins does, however, cover many of the difficulties likely to be encountered when the sample to be tested is in complex matrices, as found in most biological fluids. The rapid growth of interest in DNA based analyses has lead to the need for consistent, easy and rapid separation of oligonucleotides or polynucleotides and the potential use of HPLC is described in chapter 6. The use of HPLC in this field is likely to be superseded by HPLC of oligosaccharides from glycoproteins, an area of some interest in clinical biochemistry as knowledge of situations in which the carbohydrate side-chains of biologically active proteins are modified grows.

The use of HPLC for separation of macromolecules in clinical biochemistry is seldom encountered and this book is more likely to be of interest to biochemists in the research or industrial sectors. However, should a clinical biochemist find a need to use the technique this book is a good place to start. The chapter of mass spectrometric detectors may also be of use to the student trying to understand the intricacies of the various types of detectors now commercially available.