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Although size exclusion chromatography (SEC) is perhaps the most popular and widely used technique for determining the molecular weight distribution of polymeric materials, there have been very few texts written on this topic. During the past decade, SEC has experienced a considerable amount of growth in regard to column and detector technology and new applications. With these advances, SEC can now be used for determining absolute molecular weight, polymer chain conformation and size, and branching, as well as polymer solution properties. This book introduces the reader to the fundamentals of SEC with emphasis on practical aspects of the technique, such as column and mobile selection, calibration, new detector capabilities and guidelines for performing SEC on most types of polymers, especially those of industrial importance. This book is intended for either those new to the field of SEC, or for those research workers who require a more comprehensive background.
Documenting critical advances in this rapidly evolving field, the Second Edition highlights the need for new applications and technologies that assist in the determination of molecular weight and molecular weight distributions of polymers in an accurate, efficient manner. This volume presents the latest findings from a international team of specialists and continues to inspire and extend practical applications of size exclusion chromatography (SEC). It includes six new chapters covering high-speed size exclusion chromatography, SEC of low molecular weight materials, and the extended family of techniques, from two-dimensional liquid chromatography to high osmotic pressure chromatography.
Column Handbook for Size Exclusion Chromatography is the first comprehensive reference to provide everything one needs to know about commercial analytical and preparative columns for size exclusion and gel filtration chromatography (SEC and GFC). SEC is now widely used as a quality assurance method in the polymer industry (both synthetic and biopolymers) to determine molecular weight and molecular weight distribution. The Handbook contains contributions from every column manufacturer around the world and from many experienced column users. It covers the technology, characterization, application, evaluation, maintenance, and quality control of analytical and preparative columns for SEC and GFC. Also included are columns for two closely related techniques, hydrodynamic chromatography and high osmotic pressure chromatography.Key Features* Evaluate and select columns with confidence for specific applications* Optimize separations and improve the ruggedness of analytical methods* Extend the service time of a column* Establish a quality-control program to ensure consistency in column performance* Avoid the expense of column damage or purchases that do not give the expected results
The Second Edition of Modern Size-Exclusion Chromatography offers a complete guide to the theories, methods, and applications of size-exclusion chromatography. It provides an unparalleled, integrated, up-to-date treatment of gel permeation and gel filtration chromatography. With its detailed descriptions of techniques, data handling, compilations of information on columns and column packings, and tables of important solvents and reference materials, the book offers readers everything they need to take full advantage of this popular macromolecular characterization technique. Since publication of the first edition in 1979, there have been many important advances in the field of size-exclusion chromatography. This Second Edition brings the book thoroughly up to date, with expert coverage of: New and emerging industrial and research applications Practical aspects of size-exclusion chromatography (SEC) and multidetector and multidimensional SEC technologies for polymer architecture and copolymer analysis Updated information on the latest equipment and techniques New best practices for the lab SEC in relation to polymer characterization techniques such as GPEC, LCCC, and rheology Throughout the text, detailed examples guide you step by step through all the latest techniques and applications. With its extensive revisions and updates written by leading experts and pioneers in the field, Modern Size-Exclusion Liquid Chromatography remains the definitive resource for the broad range of researchers and scientists who use HPLC and GPC methods.
A comprehensive, practical approach to three powerful methods of polymer analysis and characterization This book serves as a complete compendium of three important methods widely used for the characterization of synthetic and natural polymers—light scattering, size exclusion chromatography (SEC), and asymmetric flow field flow fractionation (A4F). Featuring numerous up-to-date examples of experimental results obtained by light scattering, SEC, and A4F measurements, Light Scattering, Size Exclusion Chromatography and Asymmetric Flow Field Flow Fractionation takes an all-in-one approach to deliver a complete and thorough explanation of the principles, theories, and instrumentation needed to characterize polymers from the viewpoint of their molar mass distribution, size, branching, and aggregation. This comprehensive resource: Is the only book gathering light scattering, size exclusion chromatography, and asymmetric flow field flow fractionation into a single text Systematically compares results of size exclusion chromatography with results of asymmetric flow field flow fractionation, and how these two methods complement each other Provides in-depth guidelines for reproducible and correct determination of molar mass and molecular size of polymers using SEC or A4F coupled with a multi-angle light scattering detector Offers a detailed overview of the methodology, detection, and characterization of polymer branching Light Scattering, Size Exclusion Chromatography and Asymmetric Flow Field Flow Fractionation should be of great interest to all those engaged in the polymer analysis and characterization in industrial and university research, as well as in manufacturing quality control laboratories. Both beginners and experienced can confidently rely on this volume to confirm their own understanding or to help interpret their results.
There is a large and increasing variety of polymers currently in use both for domestic and industrial applications. The properties of polymers are deter mined not only by their chemical type, but also by their molecular mass and molecular mass distributions. However, while the chemical type of polymers can be determined relatively easily, the average molecular masses and molecular mass distributions are more difficult to measure. The molecular mass averages of a polymer are measured by specialized and complex techniques such as light scattering (for weight average) and osmometry (for number average). Thus, complete characterization of the molecular mass distribution of a polymer by such means requires separating the sample into many fractions which can then be examined individually. Since size exclusion chromatography was introduced as a rapid and straightforward technique for the characterization of polymer molecular mass distributions, there have been tremendous increases in development and applications, and it was felt appropriate to bring together into a single volume the information required by scientists from many disciplines who wish to use the technique. This book should be useful to existing users, those who are new to the technique, and those who may be familiar with the basic technique and now wish to extend their capabilities to more complex applications (or to consider the potential of a number of related techniques). The book will also be of general interest to the experienced liquid chromatographer.
Recent advances in the biosciences have led to a range of powerful new technologies, particularly nucleic acid, protein and cell-based methodologies. The most recent insights have come to affect how scientists investigate and define cellular processes at the molecular level. This book expands upon the techniques included in the first edition, providing theory, outlines of practical procedures, and applications for a range of techniques. Written by a well-established panel of research scientists, the book provides an up-to-date collection of methods used regularly in the authors’ own research programs.
The rapid development of new packings for aqueous size-exclusion chromatography has revolutionized this field. High resolution non-adsorptive columns now make possible the efficient separation of proteins and the rapid and precise determination of the molecular weight distribution of synthetic polymers. This technology is also being applied to the separation of small ions, the characterization of associating systems, and the measurement of branching. At the same time, fundamental studies are elucidating the mechanisms of the various chromatographicprocesses.These developments in principles and applications are assembled for the first time in this book. Fundamental issues are dealt with: the roles of pore structure and macromolecular dimensions, hydrophobic and electrostatic effects, and the determination and control of column efficiency. High-performance packings based on derivatized silica are reviewed in detail. Special techniques are thoroughly described, including SEC/LALLS, inverse exclusion chromatography, and frontal zone chromatography. Attention is focussed on special applications of size-exclusion methods, such as the characterization of micelles, separations of inorganic ions, and Hummel-Dreyer and related methods for equilibrium systems. Protein chromatography is dealt with in both dedicated sections and throughout the book as a whole.This is a particularly comprehensive and authoritative work - all the contributions review broad topics of general significance and the authors are of high repute. The material will be of special value for the characterization of synthetic water-soluble polymers, especially polyelectrolytes. Biochemists will find fundamental and practical guidance on protein separations. Researchers confronted with solutes that exhibit complex chromatographic behavior, such as humic acids, aggregating proteins, and micelles should find the contents of this volume illuminating.
The first edition of Protein Purification Protocols (1996), edited by Professor Shawn Doonan, rapidly became very successful. Professor Doonan achieved his aims of p- ducing a list of protocols that were invaluable to newcomers in protein purification and of significant benefit to established practitioners. Each chapter was written by an ex- rienced expert in the field. In the intervening time, a number of advances have w- ranted a second edition. However, in attempting to encompass the recent developments in several areas, the intention has been to expand on the original format, retaining the concepts that made the initial edition so successful. This is reflected in the structure of this second edition. I am indebted to Professor Doonan for his involvement in this new edition and the continuity that this brings. Each chapter that appeared in the original volume has been reviewed and updated to reflect advances and bring the topic into the 21st century. In many cases, this reflects new applications or new matrices available from vendors. Many of these have increased the performance and/or scope of the given method. Several new chapters have been introduced, including chapters on all the currently used protein fractionation and ch- matographic techniques. They introduce the theory and background for each method, providing lists of the equipment and reagents required for their successful execution, as well as a detailed description of how each is performed.
This book provides a clear, comprehensible and up-to-date description of how Small Angle Scattering (SAS) can help structural biology researchers. SAS is an efficient technique that offers structural information on how biological macromolecules behave in solution. SAS provides distinct and complementary data for integrative structural biology approaches in combination with other widely used probes, such as X-ray crystallography, Nuclear magnetic resonance, Mass spectrometry and Cryo-electron Microscopy. The development of brilliant synchrotron small-angle X-ray scattering (SAXS) beam lines has increased the number of researchers interested in solution scattering. SAS is especially useful for studying conformational changes in proteins, highly flexible proteins, and intrinsically disordered proteins. Small-angle neutron scattering (SANS) with neutron contrast variation is ideally suited for studying multi-component assemblies as well as membrane proteins that are stabilized in surfactant micelles or vesicles. SAS is also used for studying dynamic processes of protein fibrillation in amyloid diseases, and pharmaceutical drug delivery. The combination with size-exclusion chromatography further increases the range of SAS applications. The book is written by leading experts in solution SAS methodologies. The principles and theoretical background of various SAS techniques are included, along with practical aspects that range from sample preparation to data presentation for publication. Topics covered include techniques for improving data quality and analysis, as well as different scientific applications of SAS. With abundant illustrations and practical tips, we hope the clear explanations of the principles and the reviews on the latest progresses will serve as a guide through all aspects of biological solution SAS. The scope of this book is particularly relevant for structural biology researchers who are new to SAS. Advanced users of the technique will find it helpful for exploring the diversity of solution SAS methods and applications. Chapter 3 of this book is available open access under a CC BY 4.0 license at link.springer.com.