Analytical techniques based on separation processes, such as chromatography and electrophoresis, are finding a growing range of applications in chemical, pharmaceutical and clinical laboratories. The Wiley Separation Science Series provides the analyst in these laboratories with well focused books covering individual techniques, so that they can be applied more efficiently and effectively to contemporary analytical problems. In biotechnology, biochemistry and molecular biology, the characterization and analysis of biomolecules such as proteins, oligosaccharides and nucleic acids are of great importance. High performance liquid chromatography (HPLC) has become one of the key analytical tools in these fields, providing rapid purification and quantitative analysis of biomolecules.

High Performance Liquid Chromatography: Principles and Methods in Biotechnology covers the most important theoretical and practical aspects of the application of HPLC to the biosciences, including sample preparation, column selection, detection of biomolecules, preparative HPLC, and the use of HPLC for specific biomolecules. Finally, the technique is compared to alternative bioseparation methods. The ten chapters are written by researchers active in the biotechnology field. High Performance Liquid Chromatography: Principles and Methods in Biotechnology will be of great practical use to molecular biologists, biochemists and biotechnologists who use HPLC to purify and quantify biomolecules. It will also be of interest to those carrying out protein and DNA characterization and analysis.

Of related interest… Trace and Ultratrace Analysis by HPLC Satinder Ahuja Written by a leading scientist in the field, this monograph provides the first definitive and technically up-to-date treatment of the theory, equipment, and applications of chemistry’s most powerful reliable analytical technique. Coverage includes an encyclopedic compendium of common substances that require trace and ultratrace analysis, and features clear discussion of such important topics as considerations for HPLC equipment, sensitive detectors, sample preparation, method development, selectivity and computer-based optimizations, optimizing detectability, and much more. 1991 (0 471-51419-5) 432 pp. High Performance Liquid Chromatography in Biotechnology Edited by William S. Hancock Analytical chemists, biochemists, and chemical engineers will find this up-to-date guide to HPLC’s recent developments essential for enhancing on-the-job technical expertise. Extensive coverage includes the broad applications of HPLC, ranging from major chromatographic techniques (including reversed phase, ion exchange, affinity and hydrophobic interaction chromatography) to specific separations such as those in monoclonal antibody and nucleic acid purification. Techniques for quality control programs and advanced technology are also discussed. 1990 (0 471-82584-0) 564 pp. Unified Separation Science J. Calvin Giddings This advanced text/monograph brings together for the first time the variety of techniques used for chemical separations by outlining their common underlying mechanisms. The mass transport phenomena underlying all separation processes are developed in a simple physical-mathematical form, facilitating analysis of alternative separation techniques and the factors integral to separation power. The first six chapters provide background material applicable to a wide range of separation methods, while the final five chapters illustrate specific techniques and methods. 1991 (0 471-52089-6) 320 pp.

This book provides a detailed survey of the more powerful and less conventional HPLC detection methods available today. Addressing both theory and application, experts give their assessment of some of the most promising detection methods being developed. Long lived luminescence, near-infrared semiconductor laser fluorescence, and HPLC-mass spectrometry are among the methods discussed. In addition, an entire chapter is devoted to showing how the ultimate power of NMR may be combined with HPLC.
This book serves the needs of chromatographers who find their work restricted by the limits of conventional HPLC detection methods, and whose practical needs dictate the use of newer, more powerful methods.

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
* Multienzymesystems, 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.

Presenting the latest developments in the field for more than four decades, the Advances in Chromatography series is relied on by scientists and researchers for the most up-to-date information on a wide range of chromatographic methods and applications. Volume 52 continues this tradition with contributions by established, well-known chemists, offering cutting-edge reviews of chromatographic methods with applications in the life sciences. Featured topics include The history, development, and theory behind aerosol-based detectors Protein and peptide biomarkers that are chromatographic endpoints measured in biological fluids and tissues, such as serum, urine, and tissue biopsies Multisegment linear gradient optimization strategy in reversed-phase and hydrophilic interaction chromatographic systems Enantioselective gas chromatography performed using three distinct chiral stationary phases based on hydrogen bonding, metal ion complexation, and inclusion Analysis of the dynamic phenomena of liquid chromatographic systems with six typical reactions in the mobile phase Providing a clear presentation of topics and vivid illustrations for which this series has become known, the latest volume makes the material accessible and engaging to analytical, biochemical, organic, polymer, and pharmaceutical chemists at all levels of technical skill.

Planar Chromatography-Mass Spectrometry focuses on a relatively new approach to chemical analysis in general, and to separation science in particular. It is the first book to systemically cover the theoretical background, techniques, instrumentation, and practical applications of planar chromatography-mass spectrometry as a hyphenated tool of analytical chemistry. It also examines the high and as-yet unexploited potential of planar chromatography-mass spectrometry for analytical use in scientific investigations. This book overviews the combination of planar chromatography, a relatively simple and cost-effective separation step for determining complex mixtures of compounds, with mass spectrometry, an efficient, highly instrumental, and relatively expensive technique that enables rapid identification of separated chemical species. It covers electrophoretic-mass spectrometry methods and applications, which are considered planar chromatographic techniques and are increasingly being exploited in proteomic and molecular biology studies as well as for medical diagnostic purposes. It also provides a selection of applications, such as drug control and forensic and food analysis, including more difficult substances such as carbohydrates and lipids. The book advocates growth in using planar chromatography-mass spectrometry in laboratories that have appropriate equipment but have not yet employed the techniques in combination. It also describes the use of a relatively inexpensive commercial system that can be adopted by laboratories currently working without the coupled methodology. Aiming to improve power and efficiency when other analytical methods are inadequate, Planar Chromatography-Mass Spectrometry encourages separation science practitioners in academia and industry to combine the two methods for enhanced results.

This book describes modern preparative chromatography techniques and their application in the separation of natural products, synthetic intermediates, metabolites, macromolecules, and biologically active substances, e.g. new lead compounds from microorganisms and both marine and terrestrial higher organisms. The authors focus their book on the applications and aim to provide the reader with a rapid idea on how to obtain milligram or gram quantities for pure constituents from complex mixtures with minimum effort. Descriptions of apparatus and operation procedures together with numerous examples of actual separations - often widely scattered in literature - are provided. This completely revised second edition has been enlarged by chapters on macromolecule and and enantiomer separation and on preparative GC.

There is a growing need for high-throughput separations in food and environmental research that are able to cope with the analysis of a large number of compounds in very complex matrices. Whereas the most common approach for solving many analytical problems has often been high-performance liquid chromatography (HPLC), the recent use of fast or ultra-fast chromatographic methods for environmental and food analysis has increased the overall sample throughput and laboratory efficiency without loss (and even with an improvement) in the resolution obtained by conventional HPLC systems.This book brings together researchers at the top of their field from across the world to discuss and analyze recent advances in fast liquid chromatography-mass spectrometry (LC-MS) methods in food and environmental analysis. First, the most novel approaches to achieve fast and ultra-fast methods as well as the use of alternative and complementary stationary phases are described. Then, recent advances in fast LC-MS methods are addressed, focusing on novel treatment procedures coupled with LC-MS, new ionization sources, high-resolution mass spectrometry, and the problematic confirmation and quantification aspects in mass spectrometry. Finally, relevant LC-MS applications in food and environmental analysis such as the analysis of pesticides, mycotoxins, food packaging contaminants, perfluorinated compounds and polyphenolic compounds are described.The scope of the book is intentionally broad and is aimed at worldwide analytical laboratories working in food and environmental applications as well as researchers in universities worldwide.

An in-depth guide to HPLC column technology High-performance liquid chromatography and its derivative techniques have become the dominant analytical separation tools in the pharmaceutical, chemical, and food industries; environmental laboratories; and therapeutic drug monitoring. Although the column is the heart of the HPLC instrument and essential to its success, until now, no book has focused on the theory and practice of column technology. HPLC Columns provides thorough, state-of-the-art coverage of HPLC column technology for the practicing technician and academician alike. Along with a comprehensive discussion of the chemical and physical processes of the HPLC column, it includes fundamental principles, separation mechanisms and available technologies, column selection criteria, and special techniques. Special features include:

• Comprehensive overview of state-of-the-art HPLC column technology
• Explanation of the underlying principles of HPLC columns
• Methods for selecting columns
• Practical advice on using and applying columns, including examples
• Section by M. Zoubair El Fallah on methods development
• Special techniques, including preparative chromatography, continuous chromatography, and the simulated moving bed
• Troubleshooting section

This book covers the principles, kinetics, and applications of derivatization reactions in liquid chromatography. It emphasizes detection-oriented derivatization procedures that can be used to enhance to detectability of a wide variety of solutes using absorbance and fluorescence detection.

Many new methods directed to organic and inorganic syntheses of useful intermediates are being developed to specifically address green and sustainable chemistry principles. Highlighting the importance of green metrics, the Green Syntheses series focuses on how to reliably substantiate and validate the level of "greenness" of chemical processes, providing practical synthetic methodologies and metrics for a rigorous proof of "greenness." In Green Syntheses, Volume 1, the first book of its kind, the editors determine appropriate material efficiency green metrics and use them to compare syntheses provided by the chapter authors with those previously published. Presenting a new concept in green chemistry, this book demonstrates what future publications might look like if green principles are followed and also incorporate the important ethical aspect of supplying rigorous procedures in laboratory practice and evidence of greenness of a given synthesis protocol using metrics analysis. This inaugural volume initiates the much-needed transition from stating the 12 guiding principles on the philosophy of green chemistry to the actualization and verification of it. The book addresses primarily the issue of material efficiency metrics, which measure the amount of waste produced relative to desired product. In each contributed example, full experimental details are given showing all quantities of materials used in the procedure. Authors discuss the green merits of their protocols in conjunction with the results of a thorough metrics analysis, allowing for in-depth discussion of insights about synthesis strategy and performance characteristics of the new and prior cited plans.

This volume presents recent progress and perspectives in multi-photon processes and spectroscopy of atoms, ions, molecules and solids. The subjects in the series cover the experimental and theoretical investigations in the interdisciplinary research fields of natural science including chemistry, physics, bioscience and material science.This volume is the latest volume in a series that is a pioneer in compiling review articles of nonlinear interactions of photons and matter. It has made an essential contribution to the development and promotion of the related research fields. In view of the rapid growth in multi-photon processes and multi-photon spectroscopy, care has been taken to ensure that the review articles contained in the series are readable not only by active researchers but also those who are not yet experts but intend to enter the field.

For more than four decades, scientists and researchers have relied on the Advances in Chromatography Series for the most up-to-date information on a wide range of developments in chromatographic methods and applications. With contributions from an array of international experts, the latest volume captures new developments in this important field that yields great possibilities in a number of applications. The authors? clear presentation of topics and vivid illustrations make the material in Volume 48 accessible and engaging to biochemists and analytical, organic, polymer, and pharmaceutical chemists at all levels of technical skill.

Topics covered in this new edition include:

• The retention mechanism in reversed-phase liquid chromatography (RPLC)
• Thermodynamic modeling of chromatographic separation
• Ultra-performance liquid chromatography (ULPC)
• Biointeraction affinity chromatography
• The characterization of stationary phases in supercritical fluid chromatography with the salvation parameter model
• Silica-hydride chemistry
• Multi-dimensional gas chromatography
• Sample preparation for chromatographic analysis of environmental samples and solid-phase microextraction (SPME) with derivatization

Covering the state of the art in separation science, this volume presents timely, cutting-edge reviews on chromatography in the fields of bio-, analytical, organic, polymer, and pharmaceutical chemistry. The information contained in this latest volume will help fuel further research in this burgeoning field across the full spectrum of related disciplines.

The third edition of this guidebook provides all the basic applications needed to practice thin layer chromatography (TLC). New material includes the latest techniques on sample preparation and zone detection, the hybridization of TLC with high performance liquid chromatography (HPTLC) as it has been developed in the last few years, emphasis on numerous applications of HPTLC involving pharmaceuticals and drugs, plus the fundamental studies of mechanisms, theories and optimization of TLC.

For more than four decades, scientists and researchers have relied on the Advances in Chromatography series for the most up-to-date information on a wide range of developments in chromatographic methods and applications. For Volume 50, the series editors have invited established, well-known chemists from across the globe to offer cutting-edge reviews on their areas of expertise. The clear presentation of topics and vivid illustrations for which this series has become known makes the material accessible and engaging to analytical, biochemical, organic, polymer, and pharmaceutical chemists at all levels of technical skill.

This book provides the most up-to-date information on a wide range of developments in chromatographic methods and applications. It presents timely, cutting-edge reviews in the fields of bio-, analytical, organic, polymer, and pharmaceutical chemistry.

This book is the first example in presenting LC-MS strategies for the analysis of peptides and proteins with detailed information and hints about the needs and problems described from experts on-the-job. The best advantage is -for sure- the practical insight of experienced analysts into their novel protein analysis techniques. Readers starting in 'Proteomics' should be able to repeat each experiment with own equipment and own protein samples, like clean-up, direct protein analysis, after (online) digest, with modifications and others. Furthermore, the reader will learn more about strategies in protein analysis, like quantitative analysis, industrial standards, functional analysis and more.

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.

Established ion chromatography techniques have changed little since the 1980s but a new technique, high performance chelation ion chromatography (HPCIC), has revolutionized the area. HPCIC enables a much greater range of complex samples to be analyzed and this is the first comprehensive description of its use in the trace determination of metals. Written by world leaders in the field, it is aimed at professionals, postgraduates, chromatographers, analytical chemists, and industrial chemists. The book describes the underlying principles which give rise to the special selectivities that can be chosen for separating specific groups of metals. It also covers the latest research and gives many examples of its application to real samples. The very latest developments in detection techniques are included showing that HPCIC can rival atomic spectroscopic techniques such as ICP-MS. The detailed description of the fundamental principles controlling the separation of trace metals using chelating substrates is unique to this book. It shows how HPCIC differs from the commonly used simple ion exchange techniques and how these chelation characteristics give rise to a much more useful and versatile metal separation system. Readers will also be interested in the analysis of extremely difficult matrices, such as saturated brines, easily achieved by HPCIC but requiring very complex multi column systems using other ion chromatography methods.

High-temperature liquid chromatography has attracted much interest in recent years but has not yet recognized its full potential in the chromatographic community. There is a widespread reluctance in industry to use temperature to speed up the separation process, influence the selectivity of a separation or implement novel detection techniques. However, the technology has now matured and could revolutionize chromatography as we see it today. Better equipment, such as heating systems able to generate faster heating rates, is becoming more readily available. Also, columns based on silica gel, which can withstand higher temperatures for an extended period, are now being introduced. Nevertheless, further technological and methodical efforts are needed to establish the method in a regulated environment like the pharmaceutical industry. This is the only text to cover all the practical aspects, as well as the underlying theoretical principles, of setting up an HPLC system for high temperature operation. It is not intended solely for academics but will also benefit the researcher interested in more practical considerations. The author is a recognized expert and has conducted several studies with partners from industry to validate the method. Many real examples from these studies have been included in the book. The aim is to support practitioners in the creation of their own protocols without the need to rely solely on trial and error. The book starts with a brief definition of high temperature liquid chromatography before going on to cover: system set up; the heating system; mobile phase considerations; suitable stationary phases; method development using temperature programming; analyte stability, and special hyphenation techniques using superheated water as a mobile phase. In each chapter, experimental data is used to illustrate the main statements and the advantages over conventional HPLC are evaluated. The book concludes with a critical outlook on further developments and applications underlining the necessary advances needed to make high temperature HPLC more robust.

Analytical techniques based on separation processes, such as chromatography and electrophoresis, are finding a growing range of applications in chemical, pharmaceutical and clinical laboratories. The Wiley Separation Science Series provides the analyst in these laboratories with well-focused books covering individual techniques, so that they can be applied more efficiently and effectively to contemporary analytical problems. The different enantiomers of a drug can exhibit widely different physiological activity in degree and nature. As a result, the separation and identification of enantiomers is now a very important analytical problem and chiral chromatography is the natural technique to apply to the resolution of such mixtures. Chiral Chromatography provides the reader with a basic understanding of the nature of chromatographic separations and relates the principles specifically to the separation of enantiomers. The following information is included:

• chiral separations involving both gas and liquid chromatography
• descriptions of the apparatus used for both techniques
• detailed discussion on the retention mechanism that results in chiral selectivity
• the structure and synthesis of a wide range of chirally active stationary phases used in both gas and liquid chromatography
• preparative applications for large scale purification of enantiomers
• applications of capillary electrophoresis and capillary electrochromatography.

The porphyrins, chlorophylls, bilins and related tetrapyrroles are vital for all living organisms. Natural and synthetic tetrapyrroles are used extensively in foods, cosmetics, biotechnology, pharmaceuticals, diagnostics and medicine. Methods for their separation and characterization therefore, have a very wide area of applications. Yet, there is a dearth of books dedicated to HPLC and HPLC/MS of tetrapyrroles. Lim addresses this problem admirably by providing practical HPLC and HPLC/MS protocols coupled with in-depth chromatographic and mass spectrometric reference data. These are invaluable in the analysis, identification and characterization of porphyrins, chlorophylls, bilins and other related compounds found in biological and clinical materials. HPLC method development and optimization for coupling to mass spectrometry are also described in rich detail. Sample preparation, and suggestions for avoiding procedural artifacts during extraction of clinical and biological samples are discussed. Clinical biochemists involved in biochemical diagnosis of human porphyrias will find this monograph assuredly helpful, as would analysts, biochemists and chemists involved in the separation, isolation and characterization of natural and synthetic tetrapyrroles. Undoubtedly, Lim has contributed a master-piece containing sufficient background material for beginners and up-to-date references for all researchers in the field.

The book reviews the basic concepts and highlights the most relevant advances and developments that have taken place in the field of comprehensive two dimensional gas chromatography (GC x GC) since its introduction in 1991. The several instrumental and technical approaches assayed and developed during these seventeen years and that have contributed to the development of this powerful separation technique and to its increasing application in many areas is explained and comprehensively illustrated through a number of chapters devoted these specific topics. More specialized aspects of the technique, including theoretical aspects, modelization of the chromatographic process, software developments, and alternative couplings is also covered. Finally, special attention is paid to data treatment, for both qualitative and quantitative analysis. This book will be a practical resource that will explain from basic to specialized concepts of GC x GC and will show the current state-of-the-art and discuss future trends of this technique.
-Outlines basic concepts and principles of GCxGC technique for non-specialists to apply the technique to their research;
-Provides detailed descriptions of recent technical advances and serves as an instructional guide in latest applications in GCxGC;
-Sets the scene for possible future development and alternative new applications of technique.

Written by experts on current innovations in their fields, this 33rd volume of Advances in Chromatography covers subjects such as planar chips technology, molecular biochromatography, fusion reaction chromatography and enantioselective liquid chromatographic separations.

A concise description of models and quantitative parameters in structural chemistry and their interrelations, with 280 tables and >3000 references giving the most up-to-date experimental data on energy characteristics of atoms, molecules and crystals (ionisation potentials, electron affinities, bond energies, heats of phase transitions, band and lattice energies), optical properties (refractive index, polarisability), spectroscopic characteristics and geometrical parameters (bond distances and angles, coordination numbers) of substances in gaseous, liquid and solid states, in glasses and melts, for various thermodynamic conditions. Systems of metallic, covalent, ionic and van der Waals radii, effective atomic charges and other empirical and semi-empirical models are critically revised. Special attention is given to new and growing areas: structural studies of solids under high pressures and van der Waals molecules in gases. The book is addressed to researchers, academics, postgraduates and advanced-course students in crystallography, materials science, physical chemistry of solids.