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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.
Jump into the HPLC adventure! Three decades on from publication of the 1st German edition of Veronika Meyer's book on HPLC, this classic text remains one of the few titles available on general HPLC aimed at practitioners. New sections on the following topics have been included in this fifth edition: Comparison of HPLC with capillary electrophoresis How to obtain peak capacity van Deemter curves and other coherences Hydrophilic interaction chromatography Method transfer Comprehensive two-dimensional HPLC Fast separations at 1000 bar HPLC with superheated water In addition, two chapters on the instrument test and troubleshooting in the appendix have been updated and expanded by Bruno E. Lendi, and many details have been improved and numerous references added. A completely new chapter is presented on quality assurance covering: Is it worth the effort? Verification with a second method Method validation Standard operating procedures Measurement uncertainty Qualifications, instrument test, and system suitability test The quest for quality Reviews of earlier editions "That this text is written by an expert in both the practice and teaching of HPLC is evident from the first paragraph....not only an enjoyable, fascinating and easy read, but a truly excellent text that has and will serve many teachers, students and practitioners very well." —The Analyst “…provides essential information on HPLC for LC practitioners in academia, industry, government, and research laboratories…a valuable introduction." - American Journal of Therapeutics
The latest edition of the authoritative reference to HPLC High-performance liquid chromatography (HPLC) is today the leading technique for chemical analysis and related applications, with an ability to separate, analyze, and/or purify virtually any sample. Snyder and Kirkland's Introduction to Modern Liquid Chromatography has long represented the premier reference to HPLC. This Third Edition, with John Dolan as added coauthor, addresses important improvements in columns and equipment, as well as major advances in our understanding of HPLC separation, our ability to solve problems that were troublesome in the past, and the application of HPLC for new kinds of samples. This carefully considered Third Edition maintains the strengths of the previous edition while significantly modifying its organization in light of recent research and experience. The text begins by introducing the reader to HPLC, its use in relation to other modern separation techniques, and its history, then leads into such specific topics as: The basis of HPLC separation and the general effects of different experimental conditions Equipment and detection The column—the "heart" of the HPLC system Reversed-phase separation, normal-phase chromatography, gradient elution, two-dimensional separation, and other techniques Computer simulation, qualitative and quantitative analysis, and method validation and quality control The separation of large molecules, including both biological and synthetic polymers Chiral separations, preparative separations, and sample preparation Systematic development of HPLC separations—new to this edition Troubleshooting tricks, techniques, and case studies for both equipment and chromatograms Designed to fulfill the needs of the full range of HPLC users, from novices to experts, Introduction to Modern Liquid Chromatography, Third Edition offers the most up-to-date, comprehensive, and accessible survey of HPLC methods and applications available.
A comprehesive yet concise guide to Modern HPLC Written for practitioners by a practitioner, Modern HPLC for Practicing Scientists is a concise text which presents the most important High-Performance Liquid Chromatography (HPLC) fundamentals, applications, and developments. It describes basic theory and terminology for the novice, and reviews relevant concepts, best practices, and modern trends for the experienced practitioner. Moreover, the book serves well as an updated reference guide for busy laboratory analysts and researchers. Topics covered include: HPLC operation Method development Maintenance and troubleshooting Modern trends in HPLC such as quick-turnaround and "greener" methods Regulatory aspects While broad in scope, this book focuses particularly on reversed-phase HPLC, the most common separation mode, and on applications for the pharmaceutical industry, the largest user segment. Accessible to both novice and intermedate HPLC users, information is delivered in a straightforward manner illustrated with an abundance of diagrams, chromatograms, tables, and case studies, and supported with selected key references and Web resources. With intuitive explanations and clear figures, Modern HPLC for Practicing Scientists is an essential resource for practitioners of all levels who need to understand and utilize this versatile analytical technology.
This revision brings the reader completely up to date on the evolving methods associated with increasingly more complex sample types analyzed using high-performance liquid chromatography, or HPLC. The book also incorporates updated discussions of many of the fundamental components of HPLC systems and practical issues associated with the use of this analytical method. This edition includes new or expanded treatments of sample preparation, computer assisted method development, as well as biochemical samples, and chiral separations.
HPLC for Pharmaceutical Scientists is an excellent book for both novice and experienced pharmaceutical chemists who regularly use HPLC as an analytical tool to solve challenging problems in the pharmaceutical industry. It provides a unified approach to HPLC with an equal and balanced treatment of the theory and practice of HPLC in the pharmaceutical industry. In-depth discussion of retention processes, modern HPLC separation theory, properties of stationary phases and columns are well blended with the practical aspects of fast and effective method development and method validation. Practical and pragmatic approaches and actual examples of effective development of selective and rugged HPLC methods from a physico-chemical point of view are provided. This book elucidates the role of HPLC throughout the entire drug development process from drug candidate inception to marketed drug product and gives detailed specifics of HPLC application in each stage of drug development. The latest advancements and trends in hyphenated and specialized HPLC techniques (LC-MS, LC-NMR, Preparative HPLC, High temperature HPLC, high pressure liquid chromatography) are also discussed.
High performance liquid chromatography (HPLC) has long been recognized as one of the most useful and versatile analytical techniques. It has now progressed from being a highly expensive method of analysis to a routine technique with wide applications. Consequently there is a requirement in many chemistry and chemistry-related courses for students to acquire a detailed understanding of the principles and practice of HPLC. Written in a manner suitable for undergraduate students studying analytical chemistry and learning about chromatographic analytical techniques applied to pharmaceutical analysis, biochemistry and related disciplines, High-performance Liquid Chromatography: Fundamental Principles and Practice introduces the fundamentals of HPLC. Loosely structured in three parts, the text begins with a thorough introduction of the subject and then progresses through the essential knowledge of the instrumentation needed for HPLC. The final part covers with the applications of HPLC in real-world situations. Developed by a team of international experts from a wide cross-section of disciplines, the text is relevant to a wide range of courses.
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.
Purification and Characterization of Secondary Metabolites: A Laboratory Manual for Analytical and Structural Biochemistry provides students with working knowledge of the fundamental and advanced techniques of experimental biochemistry. Sections provide an overview of the microbiological and biochemical methods typically used for the purification of metabolites and discuss the biological significance of secondary metabolites secreted by three diverse species of bacteria. Additionally, this lab manual covers the theory and practice of the most commonly-used techniques of analytical biochemistry, UV-vis and IR spectrophotometry, high-performance liquid chromatography, mass spectrometry, X-ray crystallography and nuclear magnetic resonance, and how to evaluate and effectively use scientific data. Instructors will find this book useful because of the modular nature of the lab exercises included. Written in a logical, easy-to-understand manner, this book is an indispensable resource for both students and instructors. - Offers project lab formats for students that closely simulate original research projects - Provides instructional guidance for students to design their own experiments - Presents advanced analytical techniques - Includes access to a website with additional resources for instructors
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