Overview

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.

Full Product Details

Author:   Davy Guillarme (Geneva University, Switzerland) ,  Prof. Jean-Luc Veuthey (Geneva University, Switzerland) ,  K J Fountain ,  S Heinisch
Publisher:   Royal Society of Chemistry
Imprint:   Royal Society of Chemistry
Volume:   Volume 16
Dimensions:   Width: 15.60cm , Height: 2.50cm , Length: 23.40cm
Weight:   0.847kg
ISBN:  

9781849733885

ISBN 10:   1849733880
Pages:   465
Publication Date:   08 June 2012
Audience:   Professional and scholarly ,  College/higher education ,  Professional & Vocational ,  Tertiary & Higher Education
Format:   Hardback
Publisher's Status:   Active
Availability:   Out of stock  
The supplier is temporarily out of stock of this item. It will be ordered for you on backorder and shipped when it becomes available.

Table of Contents

1.1. Trends in HPLC; 1.2. Comparison of Chromatographic Techniques and Supports; 1.3. Constructing Kinetic Plots; 1.4. History of the Kinetic Plot Method; 1.5 Unification of the Isocratic and Gradient Kinetic Plot Method; 1.6. Relation between the Kinetic Performance under Isocratic and Gradient Elution Conditions; 1.7. Influence of the Test Conditions on the Obtained Kinetic-Performance Limit Curve; 1.8. Some Reflections on Recent Trends in Liquid Chromatography using the Kinetic Plot Method; 1.9. Conclusions; Acknowledgements; References; 2.1 UHPLC Instrumentation; 2.2 UHPLC Columns; 3.1 Introduction; 3.2 Qualitative Transfer from HPLC to UHPLC; 3.3 Normative Context for the HPLC to UHPLC Transfer; 3.4 Validation of UHPLC Methods and Equivalence of the HPLC-UHPLC Methods; 3.5 Conclusions; 3.6 References; 4.1 Introduction; 4.2 High throughput and high resolution in HT-UHPLC; 4.3 Limitations of HTLC and HT-UHPLC; 4.4 Advantage of high temperature in life science analysis; 4.5 HT-UHPLC in comprehensive on-line two-dimensional liquid chromatography (LC x LC); 4.6 Conclusion; 5. Comparison of the performance of totally porous and core-shell particles; 5.1. Introduction; 5.2. Column performance; 5.3. Possibilities of recent core-shell technology; 5.4. Particle size distribution and roughness of core-shell particles; 5.5. Loading capacity of core-shell particles; 5.6. Limited efficiency when core-shell particles packed in narrow-bore columns; 5.7. Extra column effects, contribution to band broadening; 5.8. Performance of core-shell and totally porous particles in isocratic elution mode; 5.9. Performance of core-shell and totally porous particles in gradient elution mode; 5.10. Conclusion; 6.1 Introduction; 6.2 Analytical Conditions for performing HILIC.; 6.3 Applications of HILIC in UHPLC.; 6.4 References.; 7.1 Introduction; 7.2 Selection of ionization techniques; 7.3 Overview of mass analyzers and their main features; 7.4 New developments in mass spectrometry applicable in UHPLC/MS; 7.5 Conclusions; 8.1 Introduction; 8.2 Solubility; 8.3 Ionization; 8.4 Lipophilicity; 8.5 Permeability; 8.6 Conclusion; 8.7 References; 9.1 Introduction; 9.2 UHPLC in Bioanalysis; 9.3 Sample preparation for UHPLC in bioanalysis; 9.4 Conclusions; 10.1 Introduction;10.2 GC-MS Analyses; 10.3 LC-MS(/MS) Analyses; 10.4 Application of UHPLC-MS(/MS) for Drug Testing in Sports; 10.5 Conclusion; References; 11.1 Introduction; 11.2 Use of UHPLC for the Analysis of Seized Drugs; 12.1. Pharmaceuticals as environmental contaminants; 12.2. Analysis of pharmaceuticals in environmental samples; 12.3. Occurrence of pharmaceuticals in environmental and wastewater samples; 12.4. Conclusions; Acknowledgements; References; 13.1 Introduction; 13.2 Multiple Facets of UHPLC in NP research; 13.3 Fast Targeted Analysis; 13.4 Fast Non-Targeted Analysis, Fingerprinting, and Metabolomics; 13.5 High-Resolution Profiling and Metabolite ID; 13.6 Conclusion; Acknowledgments; References; 14.1. Introduction; 14.2. Pre-analysis Considerations: Protocol Design, Sample Collection, Storage and Preparation; 14.3. Sample Preparation for Serum and/Plasma; 14.4. Collection and Storage of Urine Samples; 14.5. UHPLC-MS-Based Metabolite Profiling; 14.6. Applications of UHPLC-MS to Human Metabolic Profiling Studies; 14.7 Current Challenges for UHPLC-MS in Global metabolic profiling Studies; 14.8. Conclusions

Reviews

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. -- F. David, Research Istitute of Chromatography Chromatographia This book is the first entirely dedicated to UHPLC and related applications. -- F. Lestremau Anal. Bioanal. Chem 2012 ...the book nicely describes different fields where UHPLC has had an impact. -- F. Lestremau Anal. Bioanal. Chem 2012 ...the book is a well-balanced mix between theoretical and instrumental considerations, complemented by real life science applications. -- F. Lestremau Anal. Bioanal. Chem 2012 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. -- Frank David 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-mum 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 mum, 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. -- Francois Lestremau Anal Bioanal Chem (2012) 404:2793-2794 DOI 10.1007/s00216-012-6475-8 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. -- Frank David Chromatographia (2013) 76:577-578 DOI 10,1007/s10337-012-2357-7

This book is the first entirely dedicated to UHPLC and related applications. -- F. Lestremau * Anal. Bioanal. Chem 2012 * ...the book nicely describes different fields where UHPLC has had an impact. -- F. Lestremau * Anal. Bioanal. Chem 2012 * ...the book is a well-balanced mix between theoretical and instrumental considerations, complemented by real life science applications. -- F. Lestremau * 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. -- F. David, Research Istitute of Chromatography * Chromatographia * 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. -- Francois Lestremau * Anal Bioanal Chem (2012) 404:2793-2794 DOI 10.1007/s00216-012-6475-8 * 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. -- Frank David * Chromatographia (2013) 76:577-578 DOI 10,1007/s10337-012-2357-7 * 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. -- Frank David * Chromatographia (2013) 76:577-578 DOI 10,1007/s10337-012-2357-7 *

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. -- F. David, Research Istitute of Chromatography Chromatographia This book is the first entirely dedicated to UHPLC and related applications. -- F. Lestremau Anal. Bioanal. Chem 2012 ...the book nicely describes different fields where UHPLC has had an impact. -- F. Lestremau Anal. Bioanal. Chem 2012 ...the book is a well-balanced mix between theoretical and instrumental considerations, complemented by real life science applications. -- F. Lestremau Anal. Bioanal. Chem 2012 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. -- Frank David 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) 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. -- Francois Lestremau Anal Bioanal Chem (2012) 404:2793-2794 DOI 10.1007/s00216-012-6475-8 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. -- Frank David Chromatographia (2013) 76:577-578 DOI 10,1007/s10337-012-2357-7

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. -- F. David, Research Istitute of Chromatography * Chromatographia * This book is the first entirely dedicated to UHPLC and related applications. -- F. Lestremau * Anal. Bioanal. Chem 2012 * ...the book nicely describes different fields where UHPLC has had an impact. -- F. Lestremau * Anal. Bioanal. Chem 2012 * ...the book is a well-balanced mix between theoretical and instrumental considerations, complemented by real life science applications. -- F. Lestremau * Anal. Bioanal. Chem 2012 * 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. -- Frank David * Chromatographia (2013) 76:577-578 DOI 10,1007/s10337-012-2357-7 *

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. -- F. David, Research Istitute of Chromatography * Chromatographia * This book is the first entirely dedicated to UHPLC and related applications. -- F. Lestremau * Anal. Bioanal. Chem 2012 * ...the book nicely describes different fields where UHPLC has had an impact. -- F. Lestremau * Anal. Bioanal. Chem 2012 * ...the book is a well-balanced mix between theoretical and instrumental considerations, complemented by real life science applications. -- F. Lestremau * Anal. Bioanal. Chem 2012 * 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. -- Frank David * 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. -- Francois Lestremau * Anal Bioanal Chem (2012) 404:2793-2794 DOI 10.1007/s00216-012-6475-8 * 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. -- Frank David * Chromatographia (2013) 76:577-578 DOI 10,1007/s10337-012-2357-7 *

Author Information

Davy Guillarme gained his PhD in analytical chemistry from the University of Lyon (France) in 2004. He is now lecturer at the School of Pharmaceutical Sciences, University of Geneva (Switzerland). He is working mainly on the development of new approaches to perform ultra-fast and high resolution separations in liquid chromatography. He is also interested in the coupling of these strategies with alternative detection modes, particularly mass spectrometry. Jean-Luc Veuthey obtained his PhD in analytical chemistry from the University of Geneva (Switzerland) in 1987. He is now full professor at the School of Pharmaceutical Sciences, University of Geneva (Switzerland). His interests include the development of LC and CE hyphenated to several detection modes for the analysis of drugs and metabolites. Sample preparation and validation of the procedures are also particularly studied in his laboratory.

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