High-Performance Liquid Chromatography Methods and Applications

Author: Yu Shilin (Editor)
Publisher:
Publishing Date: 2005-06-01
Features: This book is a dedicated volume in the "Chromatography Technology Series," focusing on high-performance liquid chromatography (HPLC). In addition to providing a comprehensive and concise introduction to the classification and development of HPLC and HPLC instruments from the operator's perspective, the book emphasizes detailed explanations of various HPLC methods, including solid-phase chromatography, liquid-liquid chromatography, bonded-phase chromatography, size-exclusion chromatography, micro-column liquid chromatography, and two-dimensional HPLC. It covers separation conditions, analytical operations, and precautions for these methods. Many aspects, such as gradient elution, separation condition optimization, and experimental techniques, are summaries of the authors' and international experts' practical experiences. This book is suitable for technicians working in liquid chromatography analysis across various fields for learning and reference, and can also serve as teaching materials or reference books for students and teachers in relevant university majors.
[Preface] Preface to the Second Edition Since its publication in 2000, this book has been well-received by readers, with five reprints and sales reaching 18,000 copies. Driven by the needs of readers, it has been revised and reprinted after nearly five years. While maintaining the structure of the first edition, the second edition adds three new chapters: Gradient Elution, Micro-column Liquid Chromatography, and Two-Dimensional HPLC. The chapter on affinity chromatography from the original book has been omitted as it is planned to be published as a separate volume. Additionally, due to space constraints and the inclusion of specialized books on the applications of chromatography in life sciences, medicine, food, and environmental fields in the second edition series, the chapter on the analytical applications of HPLC in the original book has also been excluded.
To reflect the advancements in fundamental theories, new technologies, and new methods in the field of HPLC, this revision adds the following content. In the introduction and instrument sections, a brief overview of the development of HPLC methods is provided, with enhanced coverage of new high-pressure liquid pumps, guard columns, evaporative light scattering detectors, and commonly used HPLC instruments. In the sections on solid-phase chromatography, liquid-liquid chromatography, and bonded-phase chromatography, introductions to new stationary and mobile phases are included (e.g., polymer-coated silica, bonded phases with spatial protection and electrostatic shielding, monolithic columns, and superheated water mobile phases). The retention behavior of solutes in binary solvent systems and the classification methods of chemically bonded stationary phases are also briefly discussed.
Building on the understanding of solvent characteristic parameters and improving separation selectivity by altering solvent composition, the chapter on gradient elution introduces various factors affecting gradient elution [such as gradient elution time, the range of φB(）, and gradient steepness], as well as optimization methods and graphical representation techniques for gradient elution. When combined with the optimization methods for HPLC separation conditions and the use of computer software like DryLab, the process of establishing HPLC analytical methods can be significantly accelerated.
The chapter on the fundamental theories of HPLC is expanded to include an introduction to ultra-high-performance liquid chromatography (UHPLC). The section on micro-column liquid chromatography covers fundamental theories, instrument setups, micro-column preparation methods, and new technologies such as nanoliquid chromatography and ultra-high-pressure liquid chromatography. The section on two-dimensional HPLC explains performance parameters of separation systems, the technical functions of two-dimensional liquid chromatography, universal flow paths with central fractionation or full two-dimensional liquid chromatography, and their applications in proteomics research.
General steps for establishing HPLC analytical methods are supplemented with experimental techniques, including solvent purification, column packing methods, column equilibration, protection, and regeneration, gradient elution, pre-column and post-column derivatization detection, and sample preparation. The author hopes that this revised edition will not only help readers solve practical analytical problems based on fundamental principles but also keep them informed about the latest developments in new methods and technologies in the field of HPLC to meet the needs of different readers. Necessary original references are listed at the end of each chapter for further exploration.
This book has been reviewed and provided with valuable suggestions for revision by Professor Fu Rongnong, the series editor. The editor-in-chief has put in substantial effort in the publication of this book, for which the author expresses their gratitude. During this revision, Mr. Li Lang, Application Manager of Waters Company, provided samples and relevant materials from Waters' latest products, offering strong support for the revision of Chapters 2, 7, 9, and 10 of this book. Given the authors' limitations, any inaccuracies in this book are welcome to be pointed out by the readers.