HPLC in Enzymatic Analysis

HPLC in Enzymatic Analysis
Author: Edward F. Rossomando
Publisher: John Wiley & Sons
Total Pages: 480
Release: 2009-09-25
Genre: Science
ISBN: 0470111003

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The use of High Performance Liquid Chromatography (HPLC) techniques in the study of enzymatic reactions has grown significantly since the publication of the first edition of this highly successful book: the role of enzymes in biological research has expanded; the application of HPLC and enzymes has extended to more disciplines; advances in separation techniques and instrumentation have increased the capability of HPLC; and the discovery of new enzymes has spawned new methods of analysis. High Performance Liquid Chromatography in Enzymatic Analysis, Second Edition addresses these developments in its coverage of the refinements of HPLC methods and their use in a wide range of laboratory applications. It offers the same practical approach found in the first edition, incorporates a wealth of new information into existing chapters, and adds new chapters to deal with new applications, including capillary electrophoresis, forensic chemistry, microdialysis, and the polymerase chain reaction. Topics include: * Application of HPLC to the assay of enzymatic activities * Concepts and principles of HPLC, including the latest technological advances * Concepts and principles of capillary electrophoresis (CE) * Strategy for design of an HPLC/CE system for assay of enzyme activity * Preparation of enzymatic activities from tissues and single cells * Analysis of enzymatic activities in body fluids, including chromatobiosis * HPLC for the identification of new enzymatic activities * Fundamentals of the polymerase chain reaction * HPLC in forensics * Survey of enzymatic activities assayed by the HPLC method, including many new categories * Multienzyme systems, including many new examples * HPLC in the analysis of contaminated food "It is the ability of HPLC to accomplish separations completely and rapidly that led to its original application to problems in the life sciences, particularly those related to purification. An analysis of the literature revealed that this technique was used primarily for the purification of small molecules, macromolecules such as peptides and proteins, and more recently, antibodies. This application to purification has all but dominated the use of the method, and there has been a plethora of books, symposia, and conferences on the use of HPLC for these purposes. However, it was only a matter of time before others began to look beyond and to explore the possibilities that result from the capacity to make separations quickly and efficiently." --from the preface to the First Edition Easy to read and full of practical advice and hundreds of diagrams and examples, High Performance Liquid Chromatography in Enzymatic Analysis, Second Edition is an invaluable resource for students, researchers, and laboratory workers in analytical chemistry and biochemistry, molecular biology and cell biology, and for anyone interested in keeping up with this fast-growing field.

High Performance Liquid Chromatography in Enzymatic Analysis

High Performance Liquid Chromatography in Enzymatic Analysis
Author: Edward F. Rossomando
Publisher: Wiley-Interscience
Total Pages: 280
Release: 1987-05-04
Genre: Science
ISBN:

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Describes the high performance liquid chromatography (hplc) method and thoroughly explains and illustrates its use in the analysis of enzymatic activities, rather than its traditional use as a tool for separation. This change in emphasis is particularly attractive to those who wish to make use of the activity of an enzyme as an indicator of cellular function, a determinant of a given stage of differentiation (or dedifferentiation), or even as a measure of gene function. Presentation focuses on the assay of the activity of an enzyme rather than its purification. Each chapter deals with a different aspect of the method, beginning with an overview and ending with a detailed summary. The final chapter treats the growing field of assay of multienzyme systems. Includes more than 170 illustrations reproduced from the original research literature.

HPLC in Enzymatic Analysis

HPLC in Enzymatic Analysis
Author: Edward F. Rossomando
Publisher: Wiley-Liss
Total Pages: 480
Release: 1998-04-21
Genre: Science
ISBN: 9780471103400

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The use of High Performance Liquid Chromatography (HPLC) techniques in the study of enzymatic reactions has grown significantly since the publication of the first edition of this highly successful book: the role of enzymes in biological research has expanded; the application of HPLC and enzymes has extended to more disciplines; advances in separation techniques and instrumentation have increased the capability of HPLC; and the discovery of new enzymes has spawned new methods of analysis. High Performance Liquid Chromatography in Enzymatic Analysis, Second Edition addresses these developments in its coverage of the refinements of HPLC methods and their use in a wide range of laboratory applications. It offers the same practical approach found in the first edition, incorporates a wealth of new information into existing chapters, and adds new chapters to deal with new applications, including capillary electrophoresis, forensic chemistry, microdialysis, and the polymerase chain reaction. Topics include: * Application of HPLC to the assay of enzymatic activities * Concepts and principles of HPLC, including the latest technological advances * Concepts and principles of capillary electrophoresis (CE) * Strategy for design of an HPLC/CE system for assay of enzyme activity * Preparation of enzymatic activities from tissues and single cells * Analysis of enzymatic activities in body fluids, including chromatobiosis * HPLC for the identification of new enzymatic activities * Fundamentals of the polymerase chain reaction * HPLC in forensics * Survey of enzymatic activities assayed by the HPLC method, including many new categories * Multienzyme systems, including many new examples * HPLC in the analysis of contaminated food "It is the ability of HPLC to accomplish separations completely and rapidly that led to its original application to problems in the life sciences, particularly those related to purification. An analysis of the literature revealed that this technique was used primarily for the purification of small molecules, macromolecules such as peptides and proteins, and more recently, antibodies. This application to purification has all but dominated the use of the method, and there has been a plethora of books, symposia, and conferences on the use of HPLC for these purposes. However, it was only a matter of time before others began to look beyond and to explore the possibilities that result from the capacity to make separations quickly and efficiently." --from the preface to the First Edition Easy to read and full of practical advice and hundreds of diagrams and examples, High Performance Liquid Chromatography in Enzymatic Analysis, Second Edition is an invaluable resource for students, researchers, and laboratory workers in analytical chemistry and biochemistry, molecular biology and cell biology, and for anyone interested in keeping up with this fast-growing field.

Methods of Biochemical Analysis, Volume 37

Methods of Biochemical Analysis, Volume 37
Author: Clarence H. Suelter
Publisher: Wiley-Interscience
Total Pages: 324
Release: 1993-12-06
Genre: Science
ISBN: 9780471582601

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Concerned with application of special instrumental methods to problems in biology. Describes the use of x-ray crystallography in biochemistry. Reviews the application of both transmission microscopy and scanning probe microscopy to biological problems. Discusses well-developed techniques used primarily in clinical laboratories.

HPLC of Peptides and Proteins

HPLC of Peptides and Proteins
Author: Marie-Isabel Aguilar
Publisher: Springer Science & Business Media
Total Pages: 411
Release: 2008-02-03
Genre: Science
ISBN: 1592597424

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The introduction of high-performance liquid chromatography (HPLC) to the analysis of peptides and proteins some 25 years ago revolutionized the biological sciences by enabling the rapid and sensitive analysis of peptide and protein structure through the exquisite speed, sensitivity, and resolution that can be easily obtained. Today, HPLC in its various modes has become the pivotal technique in the characterization of peptides and proteins and currently plays a critical role in both our understanding of biological processes and in the development of peptide- and protein-based pharmaceuticals. The number of applications of HPLC in peptide and protein purification continues to expand at an extremely rapid rate. Solid-phase peptide synthesis and recombinant DNA techniques have allowed the production of large quantities of peptides and proteins that need to be highly purified. HPLC techniques are also used extensively in the isolation and characterization of novel proteins that will become increasingly important in the postgenomic age. The design of multidimensional purification schemes to achieve high levels of product purity further demonstrates the power of HPLC techniques not only in the characterization of cellular events, but also in the production of pepti- and protein-based therapeutics. HPLC continues to be at the heart of the analytical techniques with which scientists in both academia and in industry must arm themselves to be able to fully characterize the identity, purity, and potency of peptides and proteins.

Enzyme Assays

Enzyme Assays
Author: Robert Eisenthal
Publisher: Practical Approach
Total Pages: 310
Release: 2002
Genre: Analysis
ISBN: 9780199638208

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Enzyme assays are among the most frequently performed procedures in biochemistry and are routinely used to estimate the amount of enzyme present in a cell or tissue, to follow the purification of an enzyme, or to determine the kinetic parameters of a system. The range of techniques used tomeasure the rate of an enzyme-catalysed reaction is limited only by the nature of the chemical change and the ingenuity of the investigator. This book describes the design and execution of enzyme assays, covering both general principles and specific chapters.Building upon the highly popular first edition, this book combines revised or rewritten chapters with entirely new contributions. Topics include experimental protocols covering photometric, radiometric, HPLC, and electrochemical assays, along with methods for determining enzyme assays after gelelectrophoresis. The theory underlying each method is outlined, together with a description of the instrumentation, sensitivity and sources of error. Also included are chapters on the principles of enzyme assay and kinetic studies; techniques for enzyme extraction; high- throughout screening;statistical analysis of enzyme kinetic data; and the determination of active site concentration.This second edition of Enzyme Assays will be valuable not only to biochemists, but to researchers in all areas of the life sciences.

HPLC in Clinical Chemistry

HPLC in Clinical Chemistry
Author: Papadoyannis
Publisher: CRC Press
Total Pages: 510
Release: 1990-06-19
Genre: Science
ISBN: 9780824781392

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Based on the use of high-performance liquid chromatography (HPLC) as a means to assess quantities of separated components in HPLC detectors, this volume is structured for use by analysts who have some experience in chemical analysis or have done HPLC. It keeps theory and instrumentation at a minimum

Reaction Detection in Liquid Chromatography

Reaction Detection in Liquid Chromatography
Author: Ira S. Krull
Publisher: Routledge
Total Pages: 392
Release: 2017-10-19
Genre: Science
ISBN: 1351419757

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The first book to focus entirely on reactions for analyte detection and characterization,Reaction Detection in Liquid Chromatography depicts off- and on-line, pre- and postcolumnapproaches that have been successfully used for many classes of compounds, bothorganic and inorganic, in high performance liquid chromatography.The book gives special attention to methods and instrumentation associated with postcolumnreaction detection, discussing theory, background, principles, and equations .. .and also highlights major areas of reaction chemistry, such as immobilized ( or solution)enzymatic reactions, homogenous solution chemistry, photochemical derivation, pairedionreagents, solid phase and solid supported reagents, and reactions for inorganic species.In addition, Reaction Detection in liquid Chromatography details the efficiencies of thevarious reactions surveyed ... forecasts how the utility of each reaction is likely to be enhancedby new research ... and gives data that will allow the reader to reproducereaction-detection approaches for new analytes and samples.Reaction Detection in Liquid Chromatography is essential reading for analytical, bioanalytical,quality control, and research and development chemists. It also comprises a finereference for analysts involved in development and applications of liquid chromatographyfor specific qualitative and quantitative analyte identification; and in-house, professionalseminars.

Analytical Applications of Immobilized Enzyme Reactors

Analytical Applications of Immobilized Enzyme Reactors
Author: S. Lam
Publisher: Springer
Total Pages: 298
Release: 1994
Genre: Medical
ISBN:

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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 pro cesses 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 tech niques, which overcome certain practical, functional and economic con straints. Many natural enzymes can be stabilized by immobilization on solid matrices, with most of the activity retained, for a variety of applica tions. 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.

Advances in Chromatography

Advances in Chromatography
Author: J. Calvin Giddings
Publisher: CRC Press
Total Pages: 0
Release: 2021-07-29
Genre: Science
ISBN: 1000447170

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This book discusses the principles behind the measurements, the state of the art in technology and possible future developments in liquid chromatography (LC). It explains the limit of detection in LC for considering separately the minimum total quantity and the minimum concentration at the detector.