Analytical Applications of Immobilized Enzyme Reactors

by S. Lam (Editor), G. Malikin (Editor)

Overview

Enzymes are bio-catalysts which effect transformation of substrates to products with high specificity. The usage of enzymes in domestic and industrial applications is well known and has been well documented since the early history of civilization. With the advances in understanding of enzymology, usage of enzymes in industrial and biotechnological processes and molecular medicine has proliferated. One of the key factors in the widespread application of enzymes in modern technologies is the development of enzyme immobilization techniques, which overcome certain practical, functional and economic constraints. Many natural enzymes can be stabilized by immobilization on solid matrices, with most of the activity retained, for a variety of applications. An important application of immobilized enzymes is in liquid chromatography. In the last decade, post-column enzyme detection has become established as an important discipline in liquid chromatography. The new detection approach offers more sensitive and specific ways for measuring major classes of biomolecules. Reactors are fabricated by packing the immobilized enzymes into small columns, which can be placed immediately after an HPLC column.

Product Details

ISBN-13:	9780751400267
Publisher:	Springer Netherlands
Publication date:	09/30/1994
Edition description:	1994
Pages:	276
Product dimensions:	6.10(w) x 9.25(h) x 0.36(d)

Table of Contents

1 Immobilized enzyme reactors: developmental, practical and theoretical considerations.- 1.1 Introduction.- 1.2 Immobilized enzyme reactors (IMERs).- 1.3 IMER configurations.- 1.4 Enzyme kinetics theory.- 1.5 Flow reactor systems.- 1.6 Flow properties.- References.- 2 Flow reaction detection system designs for sugars and amino acids.- 2.1 Sugars.- 2.2 Amino acids.- 2.3 Conclusion.- References.- 3 Analysis of mono-, oligo- and polysaccharides by enzyme reaction detection.- 3.1 Introduction.- 3.2 Significance of carbohydrates.- 3.3 Flow analysis systems.- 3.4 Chromatographic detectors.- 3.5 Enzymes for carbohydrate analysis.- 3.6 Sample pretreatment.- 3.7 Applications.- Acknowledgements.- References.- 4 Uremic toxin analysis with pre- and post-column immobilized enzyme reactors.- 4.1 Introduction.- 4.2 Hypoxanthine, xanthine and uric acid.- 4.3 Creatinine, urea and electrolytes.- 4.4 Conclusion.- Acknowledgement.- References.- 5 Hydroxysteroids analysis by HPLC with semi-specific post-column immobilized enzyme reactors.- 5.1 Introduction.- 5.2 Post-column enzyme detection of hydroxysteroids.- 5.3 Preparation of the immobilized enzyme reactor.- 5.4 Characterization of the enzyme reactors.- 5.5 HPLC of hydroxysteroids.- 5.6 Enhanced chiral resolution of steroid epimers with stereospecific dehydrogenases.- References.- 6 HPLC of bile acids.- 6.1 Introduction.- 6.2 Detection of products of reaction.- 6.3 Preparation of the reactor.- 6.4 Analytical system.- 6.5 Analysis using an immobilized enzyme reactor.- 6.6 Discussion.- References.- 7 Determination of glucuronide and sulfate conjugates.- 7.1 Introduction.- 7.2 Principle of enzyme detection.- 7.3 Immobilization procedure and preparation of IMER.- 7.4 Reaction conditions and considerations.- 7.5 Selectivity, linearity and reproducibility.- 7.6 Detection of products of reaction.- 7.7 Biomedical applications.- 7.8 Conclusions.- References.- 8 Nicotinamide coenzymes: enzyme reaction detection and signal amplification by substrate recycling.- 8.1 Introduction.- 8.2 Glucose-6-phosphate dehydrogenase reactor.- 8.3 Enzyme reactor for signal amplification by substrate recycling.- References.- 9 Determination of blood ethanol with immobilized alcohol dehydrogenase and oxidase.- 9.1 Introduction.- 9.2 Alcohol dehydrogenase reaction.- 9.3 Preparation of reactors.- 9.4 Detection.- 9.5 Analysis of ethanol in blood.- 9.6 Discussion.- 9.7 Conclusion.- Acknowledgement.- References.- 10 Miscellaneous applications with immobilized enzyme reactors.- 10.1 HPLC of aldehydes with enzyme reaction detection.- 10.2 Trace analysis of zinc by immobilized metalloenzyme reactors.- References.- List of abbreviations.

Editorial Reviews

A concise reference on using immobilized enzyme reactors to improve sensitivity and selectivity in analytical chemistry. Attaching such a reactor before or after an HPLC column transforms any analyte that is a substrate of the enzyme; if the reactor is carefully designed, the products of that reaction can be made easily detectable by existing methods. Contributors from Europe, Japan, and the US explain the theory and describe the equipment and procedures, and the detection and analysis of various sugars, amino acids, steroids, and alcohols. Addressed to analytical chemists in pharmaceuticals, biochemistry, clinical chemistry, and medicine. Annotation c. Book News, Inc., Portland, OR (booknews.com)

Booknews

The editors, both experts in the field, succeeded in finding 18 co-authors who published numerous papers in the area of immobilized enzyme reactors. The result is a monograph with important practical information for those who want to use this type of chemistry in combination with liquid chromatography...a valuable tool for those developing new applications. - Journal of Chromatography; Overall, this book presents a very useful and well referenced introduction to the use of IMERs in chromatography. - Analyst; ...well written and accessible to the non-specialist...this is a well produced book, with a good number of illustration and references and a clear typeface...A useful reference book if you already use IMER technology, and equally useful if you are not a user but want to know more and get up to speed in this area. - Chromatographia

From the Publisher