HPLC and CE: Principles and Practice presents the latest information on the most powerful separation techniques available: high-performance liquid chromatography (HPLC) and capillary electrophoresis (CE). Fundamental theory, instrumentation, modes of operation, and optimization of separations are presented in a concise, non-technical style to help the user in choosing the appropriate technique quickly and accurately. Well- illustrated and containing convenientend-of-chapter summaries of the major concepts, the book provides in-depth coverage of trouble-shooting, improvement of resolution, data manipulation, selectivity, and sensitivity.
Graduate students, technicians, and researchers who must use separations with little or no background in analytical chemistry can overcome separation anxiety and get started in obtaining the best possible separations in minimal time. The book will alsobe useful to analytical chemists who need a better understanding of theory and processes.
* Fully up-to-date information on both HPLC and CE includes troubleshooting and comparisons of the two techniques
* Applicable to a wide variety of separation problems
* Covers basic concepts governing any separation as well as instrumentation and how to use it
* Helps the user to obtain optimal resolution in minimal time
* Contains information on special procedures such as chiral separations, affinity chromatography, and sample preparation
* Includes information on upcoming trends such as miniaturization
* Major concepts in each chapter are organized to allow access to information easily and quickly
* Contains practical bibliography for accessing the literature
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.
Gradient elution demystified
Of the various ways in which chromatography is applied today, few have been as misunderstood as the technique of gradient elution, which presents many challenges compared to isocratic separation. When properly explained, however, gradient elution can be less difficult to understand and much easier to use than often assumed.
Written by two well-known authorities in liquid chromatography, High-Performance Gradient Elution: The Practical Application of the Linear-Solvent-Strength Model takes the mystery out of the practice of gradient elution and helps remove barriers to the practical application of this important separation technique. The book presents a systematic approach to the current understanding of gradient elution, describing theory, methodology, and applications across many of the fields that use liquid chromatography as a primary analytical tool.
This up-to-date, practical, and comprehensive treatment of gradient elution:
* Provides specific, step-by-step recommendations for developing a gradient separation for any sample
* Describes the best approach for troubleshooting problems with gradient methods
* Guides the reader on the equipment used for gradient elution
* Lists which conditions should be varied first during method development, and explains how to interpret scouting gradients
* Explains how to avoid problems in transferring gradient methods
With a focus on the use of linear solvent strength (LSS) theory for predicting gradient LC behavior and separations by reversed-phase HPLC, High-Performance Gradient Elution gives every chromatographer access to this useful tool.
This Second Edition of the classic handbook details how to set up an HPLC system that capitalizes on the latest innovations. It covers new techniques in high-temperature, micro-flow, and ultra-fast chromatography, the linking of an HPLC to a mass spectrometer, and more. Complete with a CD-ROM and appendices, this guide has everything chromatographers need to know to confidently separate, identify, purify, and quantify compounds.
onald Armstrong updates and expands his highly praised Free Radical and Antioxidant Protocols (Humana, 1998) with a collection of new and valuable methods for evaluating the pertubations in cell function resulting from increased oxidative stress. Presented in a user-friendly, step-by-step format, these readily reproducible techniques cover both free radical-derived and antioxidant biomarkers. The methodologies demonstrated include ELISA, HPLC, infrared spectroscopy, gas chromatography-mass spectroscopy, immunoblotting, electroelution fractionation, isoelectric focusing, voltametry, and electron paramagnetic resonance imaging. Special emphasis is given to the separation of complex mixtures of plant antioxidants, soft drug design to protect from toxic oxidative metabolites, in vitro oxidation conditions, and correcting for random measurement error to improve statistical interpretation. A soon-to-be-published companion volume, Oxidants and Antioxidants: Ultrastructural and Molecular Biology Protocols (0-89603-851-3), contains state-of-the-art molecular and ultrastructural methods that expand the total number of protocols to 109 assays.
A unique approach to solving HPLC problems.
Everyone who bought "Problem Solving in HPLC" by Stavros Kromidas will equally benefit from nearly 100 new practical examples for optimization, trouble-shooting, and instrument performance given in this sequel.
The author provides
- guidance for selecting and evaluating methods, intstruments and columns,
- practical help with everyday trouble-shooting,
- advice for optimizing separations, always explaining the reason why.
In each case the problem, the solution and the conclusions are presented over a maximum of 4 pages, and in addition the book contains manufacturers' addresses, references, data tables and checklists.
Download Link ;
High pressure, or high performance, liquid chromatography (HPLC) is the method of choice for checking purity of new drug candidates, monitoring changes during scale up or revision of synthetic procedures, evaluating new formulations, and running control/assurance of the final drug product. HPLC Method Development for Pharmaceuticals provides an extensive overview of modern HPLC method development that addresses these unique concerns. Includes a review and update of the current state of the art and science of HPLC, including theory, modes of HPLC, column chemistry, retention mechanisms, chiral separations, modern instrumentation (including ultrahigh-pressure systems), and sample preparation. Emphasis has been placed on implementation in a pharmaceutical setting and on providing a practical perspective.HPLC Method Development for Pharmaceuticals is intended to be particularly useful for both novice and experienced HPLC method development chemists in the pharmaceutical industry and for managers who are seeking to update their knowledge.
* Covers the requirements for HPLC in a pharmaceutical setting including strategies for software and hardware validation to allow for use in a regulated laboratory
* Provides an overview of the pharmaceutical development process (clinical phases, chemical and pharmaceutical development activities)
* Discusses how HPLC is used in each phase of pharmaceutical development and how methods are developed to support activities in each phase
Hands-on experts from academia and industry comprehensively describe how to successfully perform all the critical HPLC techniques needed for the analysis of peptides and proteins. The methods range from commonly used techniques to those for capillary to large-scale preparative isolation. The authors have also presented a number of specific applications as case studies to illustrate the analytical approaches to a particular separation or assay challenge, with examples drawn from contemporary fields in biochemistry and biotechnology.
Development and Validation of HPLC Methods For Analytical And Preparative Purposes
Liquid chromatography (LC) is a separation method of great importance to the chemical, pharmaceutical and biotechnological industry. The principle is that a sample of a solution of the substances is injected into a column of an porous material (stationary phase) and a liquid (mobile phase) is pumped through the column. The separation of substances is based on differences in rates of migration through the column arising from different partition of the substances between the stationary and the mobile phase. Depending on the partition behavior of the different types of substances, these will elute at different times from the column outlet.