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.

UHPLC in Life Sciences

By Davy Guillarme, Jean-Luc Veuthey

The Royal Society of Chemistry

Copyright © 2012 The Royal Society of Chemistry
All rights reserved.
ISBN: 978-1-84973-388-5

Contents

Chapter 1 General Overview of Fast and High-resolution Approaches in Liquid Chromatography Deirdre Cabooter and Gert Desmet,
Chapter 2 Instrumentation and Columns for UHPLC Separations Kenneth J. Fountain and Pamela C. Iraneta,
Chapter 3 Method Transfer Between Conventional HPLC and UHPLC Benjamin Debrus, Eric Rozet, Philippe Hubert, Jean-Luc Veuthey, Serge Rudaz and Davy Guillarme,
Chapter 4 Using Elevated Temperature in UHPLC: Interest and Limitations Sabine Heinisch,
Chapter 5 Comparison of the Performance of Totally Porous and Core-shell Particles Szabolcs Fekete and Jen Fekete,
Chapter 6 Shell Particle and UHPLC Technologies for Fast Analysis of Polar Compounds in the HILIC Mode David V. McCalley,
Chapter 7 UHPLC/MS Coupling: How to Select a Suitable Configuration? Lucie Nováková, Michal Holapek, Robert Jirásko and Miroslav Lsa,
Chapter 8 UHPLC for the Determination of Physicochemical Parameters in Drug Discovery Alessandra Tania Zizzari, Pierre-Alain Carrupt and Sophie Martel,
Chapter 9 UHPLC in Modern Bioanalysis Lucie Nováková,
Chapter 10 Ultra-high Pressure Liquid Chromatography coupled to Mass Spectrometry in Doping Control Analysis F. Badoud, M. Saugy and J.-L. Veuthey,
Chapter 11 UHPLC and UHPLC-MS for the Analysis of Seized Drugs I. S. Lurie,
Chapter 12 UHPLC-MS for Multi-residue Screening of Pharmaceuticals in Environmental Samples Rebeca Lopez-Serna, Sandra Perez, Mira Petrovic and Damiá Barceló,
Chapter 13 UHPLC in Natural Products Analysis P. J. Eugster and J.-L. Wolfender,
Chapter 14 Application of UHPLC-MS to Metabolomic/metabonomic Studies in Man Helen G. Gika, Georgios A. Theodoridis and Ian D. Wilson,
Subject Index,

CHAPTER 1

General Overview of Fast and High-resolution Approaches in Liquid Chromatography

DEIRDRE CABOOTER AND GERT DESMET

Vrije Universiteit Brussel, Department of Chemical Engineering, Pleinlaan 2, 1050 Brussels, Belgium

1.1 Trends in HPLC

Over the past 50 years, column packings have evolved from irregularly shaped silica particles with sizes of 30–100 µm to spherical particles with diameters of 3–5 µm and even less than 2 µm. The tendency to develop smaller particle sizes has in essence been driven by the urge to obtain more efficient columns and faster separations. As column efficiency — especially at high velocities — mainly depends on the rate of mass transfer in the stationary phase, decreasing the diffusion distances of solutes in the stationary phase by reducing the particle size is an effective way to reach higher efficiencies. Moreover, the band spreading originating from non-uniformities in the packed bed is proportional to the particle size and will hence decrease with decreasing particle size. A drawback of using smaller particles is that the pressure drop per unit plate increases with the square of the decreasing particle size, making high operating pressures necessary to take full advantage of their performance.

Sub-2 µm particles have been developed in the last decade as an answer to the increasing demand from industry to shorten analysis times and increase sample throughput. In order to operate these sub-2 µm particles at or above their optimal flow rate, instrumentation capable of delivering pressures higher than the conventional 400 bar (up to