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UHPLC in Life Sciences : RSC Chromatography Monographs - Davy  Guillarme

UHPLC in Life Sciences

By: Davy Guillarme (Editor), Jean-Luc Veuthey (Editor)

Hardcover | 8 January 2012

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Since its commercial introduction in 2004, UHPLC (Ultra-High Performance Liquid Chromatography) has begun to replace conventional HPLC in academia and industry and interest in this technique continues to grow. Both the increases in speed and resolution make this an attractive method; particularly to the life sciences and more than 1500 papers have been written on this strongly-evolving topic to date. This book provides a solid background on how to work with UHPLC and its application to the life sciences. The first part of the book covers the basics of this approach and the specifics of a UHPLC system, providing the reader with a solid background to working properly with such a system. The second part examines the application of UHPLC to the life sciences, with a focus on drug analysis strategies. UHPLC-MS, a key technique in pharmaceutical and toxicological analyses, is also examined in detail. The editors (Davy Guillarme and Jean-Luc Veuthey) were some of the earliest adopters of UHPLC and have published and lectured extensively on this topic. Between them they have brought together an excellent team of contributors from Europe and the United States, presenting a wealth of expertise and knowledge. This book is an essential handbook for anyone wishing to adopt an UHPLC system in either an academic or industrial setting and will benefit postgraduate students and experienced workers alike. This unique book is dedicated to the basics of UHPLC, a current "hot topic", and its application in the field of drug analysis. Since its commercial introduction in 2004, there has been a tremendous interest in UHPLC (in both academic laboratories and industries) to speed up and increase resolution of traditional HPLC. The goal of this book is to provide a solid background on how to work properly in UHPLC and how this strategy has been applied for analysis in the life sciences. The first half of the book covers many of the specificities of UHPLC, thus providing a guideline for working properly in UHPLC. The second part is dedicated to the analysis of drugs with UHPLC and above all UHPLC-MS, which is a field of high interest for pharmaceutical and toxicological analysis.
Industry Reviews

The book gives a very comprehensive overview of the fundamentals and applications of ultra high pressure liquid Chromatography. In the last decade, particularly after the commercial introduction of an ''UPLC'' system in 2004, the application of ultra high pressures ([400 bar) in HPLC has resulted in a major step forward, resulting in very high resolution separations and/or very high throughput methods. Using sub-two micron particles in HPLC has, however, several consequences for method development and, especially for comparing UHPLC methods, performed on dedicated equipment, with classical methods performed at 400 bar. The editors of this book have made a great effort to cover all aspects of UHPLC and its application in life sciences.

Starting from the fundamentals, Chapter 1 explains how UHPLC systems can be compared to classical HPLC

separations, based on the concept of kinetic plots. These kinetic plots describe the ''working domain'' of the

combination of an instrument with maximum pressure specifications, with a column length, particle size, and

mobile phase viscosity. This approach is being used more and more for comparing different HPLC systems in terms of maximum analysis speed (analysis time) as a function of maximum plate number (efficiency). Kinetic plots can be used for isocratic and gradient methods, comparison of porous and core-shell particles, etc. In Chapter 2, adescription of the critical aspects of instrumentation is given. Unfortunately, this chapter is a little too focused on one specific commercial instrument, highlighting its advantages, while the advantages of other systems are less discussed. Chapter 3 discusses the factors to be taken into account when transferring classical methods into UHPLC methods and the consequences for method validation (e.g., in pharmaceutical methods) are highlighted. The last part of this chapter is especially interesting for method developers in pharmaceutical labs. In Chapter 4, attention is paid to the role of elevated temperature as an alternative to, or in combination with UHPLC. The possibilities and limitations of core-shell column technology and of hydrophilic interaction

liquid chromatography (HILIC) are discussed in the next two chapters. To complete the first part of the book,

Chapter 7 gives a clear overview of state-of-the-art mass spectrometers used in hyphenation with HPLC, and some critical comments are made on their combination with UHPLC. The second part of the book gives a very broad overview of the applications of UHPLC that goes beyond the life sciences. Applications in drug discovery, bioanalysis, doping control, analysis of drugs of abuse, natural product analysis and metabolomics are discussed in these chapters. In my opinion, some authors tried to be too comprehensive (for instance, on the description of sample preparation techniques for bioanalysis in Chapter 9), and this results in too much information that is often less significant for UHPLC. However, most of the examples given in the

different chapters clearly illustrate the potential of UHPLC and an extensive literature overview is included in each chapter.

This book, Volume 16 in the well-known series of RSC chromatography monographs, also has a clear layout, good quality of figures and can be recommended to both students of separation science and professionals using UHPLC.

* Chromatographia (2013) 76:577-578 DOI 10,1007/s10337-012-2357-7 *

This book, part of the RSC chromatographymonographs series, is dedicated to the advantages provided by the introduction of ultra-high-pressure instrumentation in the liquid chromatography field and focuses on selected applications in life sciences. Liquid chromatography is one of the most widely used separation techniques. The development of ultra-high pressure liquid (UHPLC) instruments, able to deliver (well) over 400 bar and therefore to use sub-2-?m columns, provided to perform analyses faster and obtain improvedresolution. This has contributed to the widespread use of UHPLC technology, which has now been largely adopted by both academia and industry. Contents This book has 14 chapters. Chapters 1-7 focus on the theoretical basis of UHPLC. Chapter 1 explains, via the kinetic plot representation, the advantages provided by use of high pressure for improving the speed or resolution of analysis. Various examples considering different column technologies (sub-2 ?m, core-shell) or particle sizes are presented. Chapter 2 is dedicated to the technical characteristics of ultra-high-pressure instrumentation, notably the importance of extra-column volume and the use of dedicated columns. The transfer of the method between highperformance liquid chromatography and UHPLC is described in Chap. 3, covering inherent issues such as selectivity changes. High temperature in combination with UHPLC is presented in Chap. 4, notably describing how to take advantage of the reduction of mobile phase viscosity for increased resolution or two-dimensional liquid chromatography. A general description of core-shell particles is given in Chap. 5, and applications to UHPLC instrumentation presented. The use of hydrophilic liquid chromatography (HILIC) for the analysis of polar compounds is explained in Chap. 6 and is illustrated by examples of HILIC separation in UHPLC mode. Chapter 7 presents the coupling of UHPLC with mass spectrometry. The various type of mass spectrometers (quadrupole, ion trap, Orbitrap, time of flight, etc.) and their combinations (triple

quadrupole/quadrupole time of flight, etc.) are reviewed and their compatibility with the speed provided by UHPLC are discussed. Chapters 8-14 focus on specific applications in various fields related to life science analysis. Determination of physicochemical parameters by UHPLC such as pKa and lipophilicity in the drug discoveryarea is described in Chap. 8. In Chap. 9, the combination of UHPLC and various mass spectrometers for bioanalysis is presented. Chapter 10 deals with doping control analysis and in particular urine analysis and coupling with mass spectrometry. Strategies for rapid screening of drugs of abuse are discussed in Chap. 11. Application of UHPLC to environmental analysis and in particular to the analysis of pharmaceutical residues in water is presented in Chap. 12. Chapter 13 deals with natural product analysis. UHPLC has enabled researchers to obtain higher resolution, leading to better defined fingerprint patterns and better profiling for metabolite identification of plant constituents. Finally, the use of UHPLC for metabolomic/metabonomic studies in humans is discussed in Chap. 14 and in particular applications to human metabolic profiling studies and biomarker determination in biological fluids and tissues. Comparison with existing literature This book is the first entirely dedicated to UHPLC and related applications. The company Waters edited Beginners guide to UPLC: ultraperformance liquid chromatography but this book, whose chapters have been written by experts in their particular fields, provides a more general scope and application range. Critical assessment The book encompasses the various aspect of UHPLC by first describing the fundamental advantages but also the specificity of this technology. The latest developments and main issues related to UHPLC usesuch as method transfer are discussed. The latest column technologies such as core-shell particles and HILIC are thoroughly described. However, frictional heating, one of the main issues related to the use of ultrahigh pressure, is only briefly mentioned and could have been explained in more depth in the theoretical section. The different applications selected highlight the main advantages that UHPLC has provided in terms of either high throughput or high resolution. Overall, the book nicely describes different fields where UHPLC has had an impact in the last few years, demonstrating the general contribution and application of the technique. The compatibility of UHPLC with the various types of detection (UV and particularly mass spectrometry) is extensively explained. All chapters relate to relevant work published in the literature, which can lead the reader to find out more technical details. Summary UHPLC in life sciences is a monograph which provides a comprehensive description of aspects of UHPLC. Overall, the book is a well-balanced mix between theoretical and instrumental considerations, complemented by real life science applications. The book is addressed to members of both academia and industry who would like to learn about UHPLC and about the advantage it can provide in life science analysis. Liquid chromatography users should be the most interested audience, and the book is not recommended for undergraduate students lacking practical experience in this field.

* Anal Bioanal Chem (2012) 404:2793-2794 DOI 10.1007/s00216-012-6475-8 *

The editors of this book have made a great effort to cover all aspects of UHPLC and its application in life sciences

This book, Volume 16 in the well-known series of RSC chromatography monographs, also has a clear layout, good quality of figures and can be recommended to both students of separation science and professionals using UHPLC.

* Chromatographia (2013) 76:577-578 DOI 10,1007/s10337-012-2357-7 *

This book is the first entirely dedicated to UHPLC and related applications.

* Anal. Bioanal. Chem 2012 *

...the book nicely describes different fields where UHPLC has had an impact.

* Anal. Bioanal. Chem 2012 *

...the book is a well-balanced mix between theoretical and instrumental considerations, complemented by real life science applications.

* Anal. Bioanal. Chem 2012 *

"The book gives a very comprehensive overview of the fundamental and application of ultra high pressure liquid chromatography."

"... has a clear layout, good quality of figures and can be recommended to both students of separation science and professionals using UHPLC."

"... most of the examples given in the different chapters clearly illustrate the potential of UHPLC..."

"... an extensive literature overview is included in each chapter."

* Chromatographia *

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