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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.
Essentials in Modern HPLC Separations, Second Edition discusses the role of separation in high performance liquid chromatography (HPLC). This new and updated edition systematically presents basic concepts as well as new developments in HPLC. Starting with a description of basic concepts, it provides important guidance for the practical utilization of various HPLC procedures, such as the selection of the HPLC type, proper choice of the chromatographic column, selection of mobile phase and selection of the method of detection, all of which are in correlation with the physico-chemical characteristics of the compounds separated. Every chapter has been carefully reviewed, with several new sections added to bring the book completely up-to-date. Hence, it is a valuable reference for students and professors in chemistry. - Provides a thoroughly updated resource, with an entirely new section on Computer-aided Method Development in HPLC and new subsections on miniaturization and automation in HPLC, chemometric aspects of HPLC, green solvent use in HPLC, and more - Includes insights into the chromatographic process to find the optimum solution for analyzing complex samples - Presents a basis for understanding the utilization of modern HPLC for applications, particularly for the analysis of pharmaceutical, biological, food, beverage and environmental samples
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
Techniques in Protein Chemistry III compiles papers presented at the Fifth Protein Society Symposium in Baltimore on June 22-26, 1991. This book discusses the protein and peptide recovery from PVDF membranes; high-sensitivity peptide mapping utilizing reversed-phase microbore and microcolumn liquid chromatography; and capillary electrophoresis for preparation of peptides and direct determination of amino acids. The TFMSA/TFA cleavage in t-Boc peptide synthesis; applications of automatic PTC amino acid analysis; and identification of O-glycosylation sites with a gas phase sequencer are also elaborated. This text likewise covers the conformational stability of the molten globule of cytochrome c and role of aqueous solvation in protein folding. This publication is useful to students and researchers interested in methods and research approaches on protein chemistry.
Separation Methods in Drug Synthesis and Purification
While working as a chromatographer in the pharmaceutical industry, it became apparent to the editor that there was a pressing need for a comprehensive reference text for analysts working on the resolution of enantiomers by liquid chromatography (LC). This need arises from the fact that, whereas previously it was very difficult to determine enantiomers by direct means, there is now a wide choice of direct LC methods. At the same time, regulatory authorities have been changing their attitudes towards the administration of pharmaceuticals as racemates, partly because it is now possible to study the individual enantiomers. Clearly this abundance of new information needs to be rationalized. More importantly, the chiral LC systems which are commercially available or readily accessible to the practising chromatographer needed to be reviewed and, to a much greater extent than in existing reviews or books, discussed in terms of their practical application. Accordingly this book is very much orientated towards the practical aspects of these commercially available and readily accessible chiral LC systems. To this end, it is written for practising chromatographers by a team of practising, experienced chromatographers who have spent many years tackling the problems presented by resolving enantiomers by LC. The practical aspects of common chiral LC systems cannot be fully understood if discussed in isolation.
A comprehensive guide to smart materials and how they are used in sample preparation, analytical processes, and applications This comprehensive, two-volume handbook provides detailed information on the present state of new materials tailored for selective sample preparation and the legal frame and environmental side effects of the use of smart materials for sample preparation in analytical chemistry, as well as their use in the analytical processes and applications. It covers both methodological and applied analytical aspects, relating to the development and application of new materials for solid-phase extraction (SPE) and solid-phase microextraction (SPME), their use in the different steps and techniques of the analytical process, and their application in specific fields such as water, food, air, pharmaceuticals, clinical sciences and forensics. Every chapter in Handbook of Smart Materials in Analytical Chemistry is written by experts in the field to provide a comprehensive picture of the present state of this key area of analytical sciences and to summarize current applications and research literature in a critical way. Volume 1 covers New Materials for Sample Preparation and Analysis. Volume 2 handles Analytical Processes and Applications. Focuses on the development and applications of smart materials in analytical chemistry Covers both, methodological and applied analytical aspects, for the development of new materials and their use in the different steps and techniques of the analytical process and their application in specific fields Features applications in key areas including water, air, environment, pharma, food, forensic, and clinical Presents the available tools for the use of new materials suitable to aid recognition process to the sample preparation and analysis A key resource for analytical chemists, applied laboratories, and instrument companies Handbook of Smart Materials in Analytical Chemistry, 2V Set is an excellent reference book for specialists and advanced students in the areas of analytical chemistry, including both research and application environments.
Ion-Pair Chromatography (IPC) is a rapidly evolving method for difficult analyses of organic and inorganic ions and ionogenic, neutral, and zwitterionic compounds. The possibilities for this technology continue to grow as novel ion-pair reagents and strategies are introduced at an accelerated level. Compensating for a dearth in the literature, Ion-
Liquid Chromatography: Applications, Second Edition,is a single source of authoritative information on all aspects of the practice of modern liquid chromatography. It gives those working in both academia and industry the opportunity to learn, refresh, and deepen their knowledge of the wide variety of applications in the field. In the years since the first edition was published, thousands of papers have been released on new achievements in liquid chromatography, including the development of new stationary phases, improvement of instrumentation, development of theory, and new applications in biomedicine, metabolomics, proteomics, foodomics, pharmaceuticals, and more. This second edition addresses these new developments with updated chapters from the most expert researchers in the field. - Emphasizes the integration of chromatographic methods and sample preparation - Explains how liquid chromatography is used in different industrial sectors - Covers the most interesting and valuable applications in different fields, e.g., proteomic, metabolomics, foodomics, pollutants and contaminants, and drug analysis (forensic, toxicological, pharmaceutical, biomedical) - Includes references and tables with commonly used data to facilitate research, practical work, comparison of results, and decision-making
This book is intended to familiarize biochemists with HPLC. Theoretical aspects of each mode of chromatography are discussed in chapters 1-9, providing an understanding of the various modes of chromatography which are now possible using commercially available columns, from reversed phase to affinity. Practical aspects and instrumentation are covered in chapter 10. The bulk of the book, which follows, presents examples and applications of each mode of chromatography in current biochemical practice.