Hydrophilic Interaction Chromatography E-Book

Bernard A. Olsen

• Herausgeber: John Wiley & Sons
• Kategorie: Wissenschaft und neue Technologien
• Serie: Chemical Analysis: A Series of Monographs on Analytical Chemistry and Its Applications
• Sprache: Englisch

Seitenzahl: 518

Veröffentlichungsjahr: 2012

Beschreibung

Discover how to use HILIC to analyze and better understand polar compounds An increasingly popular analytical method, hydrophilic interaction chromatography (HILIC) has the ability to retain and separate polar compounds that are often difficult to analyze by reversed-phase high-performance liquid chromatography (HPLC) or other analytical methods. Offering a comprehensive review, this book enables readers to develop a fundamental understanding of how HILIC works and then apply that knowledge to develop and implement a variety of practical applications. Hydrophilic Interaction Chromatography begins with discussions of HILIC retention mechanisms, stationary phases, and general method development. This sets the foundation for the book's extensive coverage of applications. The authors address unique separation challenges for bioanalytical, environmental, pharmaceutical, and biochemical applications. Moreover, there is a thorough discussion of HILIC in two-dimensional chromatography. With contributions from leading analytical scientists who have extensive experience in HILIC as well as HPLC, Hydrophilic Interaction Chromatography serves as a practical guide for researchers, featuring: * Detailed examples of HILIC methods and development approaches * Thorough explanations of retention mechanisms and the impact of stationary phase and mobile phase properties on separations * Step-by-step guidance for developing efficient, sensitive, and robust HILIC methods * References to the primary literature at the end of each chapter Hydrophilic Interaction Chromatography is written for scientists who use or develop analytical methods for the separation of polar compounds. In particular, these researchers will discover how HILIC can be used to analyze and better understand the composition of pharmaceutical, bioanalytical, biochemical, chemical, food, and environmental samples.

Leseprobe

Table of Contents

COVER

CHEMICAL ANALYSIS

TITLE PAGE

COPYRIGHT PAGE

PREFACE

CONTRIBUTORS

CHAPTER 1 SEPARATION MECHANISMS IN HYDROPHILIC INTERACTION CHROMATOGRAPHY

1.1 INTRODUCTION

1.2 HISTORICAL BACKGROUND: RECOGNITION OF THE CONTRIBUTION OF PARTITION, ION EXCHANGE, AND RP INTERACTIONS TO THE RETENTION PROCESS

1.3 RECENT STUDIES ON THE CONTRIBUTORY MECHANISMS TO HILIC RETENTION

1.4 CONCLUSIONS

CHAPTER 2 STATIONARY PHASES FOR HILIC

2.1 INTRODUCTION

2.2 HILIC STATIONARY PHASES

2.3 COMMERCIAL HILIC PHASES

2.4 CONCLUSIONS

ACKNOWLEDGMENTS

CHAPTER 3 HILIC METHOD DEVELOPMENT

3.1 INTRODUCTION

3.2 GENERAL METHOD DEVELOPMENT CONSIDERATIONS

3.3 HILIC METHOD DEVELOPMENT

3.4 DETECTION FOR HILIC METHODS

3.5 CONCLUSIONS

CHAPTER 4 PHARMACEUTICAL APPLICATIONS OF HYDROPHILIC INTERACTION CHROMATOGRAPHY

4.1 INTRODUCTION

4.2 DETERMINATION OF COUNTERIONS

4.3 MAIN COMPONENT METHODS

4.4 DETERMINATION OF IMPURITIES

4.5 EXCIPIENTS

4.6 CHIRAL APPLICATIONS

4.7 CONCLUSIONS

CHAPTER 5 HYDROPHILIC INTERACTION CHROMATOGRAPHY (HILIC) FOR DRUG DISCOVERY

5.1 DRUG DISCOVERY MODEL

5.2 HILIC APPLICATIONS FOR IN VITRO BIOLOGY

5.3 HILIC APPLICATIONS FOR DISCOVERY CHEMISTRY

5.4 PRACTICAL CONSIDERATIONS

5.5 CONCLUSIONS

CHAPTER 6 ADVANCES IN HYDROPHILIC INTERACTION CHROMATOGRAPHY (HILIC) FOR BIOCHEMICAL APPLICATIONS

6.1 INTRODUCTION

6.2 CARBOHYDRATES

6.3 NUCLEOBASES AND NUCLEOSIDES

6.4 OLIGONUCLEOTIDES

6.5 AMINO ACIDS AND PEPTIDES

6.6 PROTEINS

6.7 PHOSPHOLIPIDS

6.8 CONCLUSIONS

CHAPTER 7 HILIC-MS FOR TARGETED METABOLOMICS AND SMALL MOLECULE BIOANALYSIS

7.1 INTRODUCTION

7.2 THE ROLE OF HILIC-MS IN TARGETED METABOLOMICS VERSUS OTHER LC MODES

7.3 STRATEGIES FOR METHOD DEVELOPMENT BASED ON RETENTION BEHAVIOR OF TARGETED METABOLITES ON HILIC STATIONARY PHASES

7.4 SUMMARY

ACKNOWLEDGMENTS

CHAPTER 8 HILIC FOR FOOD, ENVIRONMENTAL, AND OTHER APPLICATIONS

8.1 INTRODUCTION

8.2 FOOD APPLICATIONS FOR HILIC

8.3 ENVIRONMENTAL AND OTHER APPLICATIONS OF HILIC

8.4 CONCLUSIONS

CHAPTER 9 THEORY AND PRACTICE OF TWO-DIMENSIONAL LIQUID CHROMATOGRAPHY SEPARATIONS INVOLVING THE HILIC MODE OF SEPARATION

9.1 FUNDAMENTALS OF MULTIDIMENSIONAL LIQUID CHROMATOGRAPHY

9.2 COMPLEMENTARITY OF HILIC SELECTIVITY TO OTHER SEPARATION MODES

9.3 INSTRUMENTATION AND EXPERIMENTAL CONSIDERATIONS

9.4 APPLICATIONS

9.5 THE FUTURE OF HILIC SEPARATIONS IN 2DLC

INDEX

Cover design: John Wiley & Sons, Inc.

Cover illustration: Copyright Brian W. Pack

Copyright © 2013 by John Wiley & Sons, Inc. All rights reserved

Published by John Wiley & Sons, Inc., Hoboken, New Jersey

Published simultaneously in Canada

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Library of Congress Cataloging-in-Publication Data:

Olsen, Bernard A., 1953– author.

 Hydrophilic interaction chromatography : a guide for practitioners / Bernard A. Olsen, Brian W. Pack.

pages cm

 Includes bibliographical references and index.

 ISBN 978-1-118-05417-8 (hardback)

 1. Hydrophilic interaction liquid chromatography. I. Pack, Brian W., 1970– author. II. Title.

 QD79.C454047 2013

 543'.84–dc23

2012027157

PREFACE

The popularity of hydrophilic interaction chromatography (HILIC) has grown rapidly in recent years. The HILIC mode can provide retention and separation of polar compounds that are difficult to analyze by reversed-phase high-performance liquid chromatography (RP-HPLC) or other means. HILIC has been utilized for a wide variety of applications including drugs and metabolites in biological fluids, biochemicals, pharmaceuticals (from drug discovery to quality control), foods, and environmental. Multiple HILIC stationary phases have been developed, and methods employing mass spectrometric detection or HILIC as part of two-dimensional separation systems are becoming more common.

Many researchers do not have an extensive background or experience with HILIC, particularly as compared with RP-HPLC. Several established references are available for information on RP-HPLC theory, mechanisms, and method development. Despite the recent growth in the use of HILIC, less information is available to guide potential practitioners in the understanding and development of robust HILIC separations. The lack of familiarity with HILIC can lead to trial-and-error method development and perhaps less-than-optimal results for a given application.

We sought to compile a book that would be a good reference for HILIC fundamentals as well as to provide a broad overview of popular areas of application. Our goal was to provide a resource for several important topics to those who want to explore HILIC as a separation mode. We believe that a basic understanding of retention mechanisms and the impact of stationary phase and mobile phase properties on separations can lead to more efficient and effective development of robust separation methods. The first three chapters of the book are devoted to HILIC retention mechanisms, stationary phases, and general aspects of method development. These chapters provide a foundation for subsequent chapters dealing with different areas of application. The application chapters focus on specific areas of interest to workers in the respective fields being addressed. Unique separation challenges are presented for bioanalytical, environmental, pharmaceutical, and biochemical applications, as well as a thorough discussion of HILIC in two-dimensional chromatography. Illustrative examples of several HILIC methods and development approaches are highlighted, and references for further details are provided.

We are indebted to all the authors who contributed to the book. We believe they provided discussions of their subject area in a concise fashion with minimal redundancy with the other chapters. Research to understand the fundamentals of HILIC separations and further application of HILIC to analytical problems will certainly continue. Our goal is that this book will be a useful reference for current and future HILIC practitioners.

We are also grateful to Dr. Mark F. Vitha, Chemical Analysis Series Editor, for his valuable help throughout the preparation of the book.

BERNARD A. OLSENBRIAN W. PACKNovember 7, 2013

CONTRIBUTORS

Alfonso Espada, Analytical Technologies Department, Centro de Investigacion Lilly S.A., Madrid, Spain

Aikaterini M. Gremilogianni, Laboratory of Analytical Chemistry, Department of Chemistry, University of Athens, Athens, Greece

Stephen R. Groskreutz, Department of Chemistry, Gustavus Adolphus College, St. Peter, MN

Yong Guo, School of Pharmacy, Fairleigh Dickinson University, Madison, NJ

Mohammed E.A. Ibrahim, Department of Chemistry, University of Alberta, Gunning/Lemieux Chemistry Centre, Edmonton, Alberta, Canada

Michael A. Koupparis, Laboratory of Analytical Chemistry, Department of Chemistry, University of Athens, Athens, Greece

Charles A. Lucy, Department of Chemistry, University of Alberta, Gunning/Lemieux Chemistry Centre, Edmonton, Alberta, Canada

Michelle L. Lytle, Analytical Sciences Research and Development, Lilly Research Laboratories, A Division of Eli Lilly and Company, Indianapolis, IN

David V. McCalley, Department of Applied Sciences, University of the West of England, Bristol, UK

Nikolaos C. Megoulas, Laboratory of Analytical Chemistry, Department of Chemistry, University of Athens, Athens, Greece

Hien P. Nguyen, Department of Chemistry and Biochemistry, The University of Texas at Arlington, Arlington, TX

Bernard A. Olsen, Olsen Pharmaceutical Consulting, LLC, West Lafayette, IN

Brian W. Pack, Analytical Sciences Research and Development, Lilly Research Laboratories, A Division of Eli Lilly and Company, Indianapolis, IN

Fred Rabel, ChromHELP, LLC, Woodbury, NJ

Donald S. Risley, Pharmaceutical Sciences Research and Development, Lilly Research Laboratories, A Division of Eli Lilly and Company, Indianapolis, IN

Kevin A. Schug, Department of Chemistry and Biochemistry, The University of Texas at Arlington, Arlington, TX

V. Scott Sharp, Pharmaceutical Sciences Research and Development, Lilly Research Laboratories, A Division of Eli Lilly and Company, Indianapolis, IN

Dwight R. Stoll, Department of Chemistry, Gustavus Adolphus College, St. Peter, MN

Mark Strege, Analytical Sciences Research and Development, Lilly Research Laboratories, A Division of Eli Lilly and Company, Indianapolis, IN

Heather D. Tippens, Department of Chemistry and Biochemistry, The University of Texas at Arlington, Arlington, TX

Xiande Wang, Johnson & Johnson, Raritan, NJ

CHAPTER 1

SEPARATION MECHANISMS IN HYDROPHILIC INTERACTION CHROMATOGRAPHY

DAVID V. MCCALLEY

Department of Applied Sciences, University of the West of England, Bristol, UK

1.1 Introduction

1.2 Historical Background: Recognition of the Contribution of Partition, Ion Exchange, and RP Interactions to the Retention Process

1.3 Recent Studies on the Contributory Mechanisms to HILIC Retention

1.3.1 Overview

1.3.2 Contribution of Adsorption and Partition to HILIC Separations

1.3.3 Further Studies on the Contribution of Ionic Retention in HILIC

1.3.4 RP Retention on Bare Silica

1.3.5 Electrostatic Repulsion Hydrophilic Interaction Chromatography (ERLIC): A New Separation Mode in HILIC

1.4 Conclusions

1.1 INTRODUCTION

Hydrophilic interaction chromatography (HILIC) is a technique that has become increasingly popular for the separation of polar, hydrophilic, and ionizable compounds, which are difficult to separate by reversed-phase chromatography (RP) due to their poor retention when RP is used. HILIC typically uses a polar stationary phase such as bare silica or a polar bonded phase, together with an eluent that contains at least 2.5% water and >60% of an organic solvent such as acetonitrile (ACN). However, these values should not be regarded as definitive of the rather nebulous group of mobile and stationary phase conditions that are considered to constitute HILIC. Figure 1.1 shows the number of publications on HILIC between the years 1990 (when the term was first employed) and 2010 according to the Web of Knowledge [1] using the search terms “HILIC” or “hydrophilic interaction (liquid) chromatography.” For the first 12 years or so, the number of publications remained between 1 and 15, but after this period, interest increased rapidly from 19 publications in 2003 to 267 in 2010. While HILIC has unique retention characteristics for hydrophilic compounds, this increase in interest also reflects the advantages of HILIC over RP methods in situations where either technique is applicable. These advantages result mostly from the high organic content of typical mobile phases and their resultant high volatility and low viscosity. A particular advantage is in coupling HILIC to mass spectrometry (MS) as mobile phases are more efficiently desolvated in interfaces such as electrospray, giving rise to better sensitivity than with RP methods. Thus, Grumbach and coworkers demonstrated sensitivity increases of 3–4 orders of magnitude when comparing the analysis of the drugs salbutamol and bamethan by HILIC on a bare silica column using a gradient analysis starting at 90% ACN with that on a C18 RP column using a gradient starting at 0% ACN [2]. Columns can be used at considerably lower pressures than in RP; the viscosity of 80–90% ACN mixtures with water as typically used in HILIC is only about half that of 20–30% ACN mixtures that might be used in RP separations [3]. Alternatively, longer columns can be used at pressures typically found in RP analysis, allowing high efficiencies to be obtained [4]. For example, when combining the low viscosity of HILIC with the efficiency gains shown by superficially porous (shell) particle columns, it is possible to generate column efficiencies in excess of 100,000 plates with reasonable analysis times, and using pressures that are well within the capabilities of conventional HPLC systems (pressure < < 400 bar). Low viscosity also results in increased solute diffusion in the mobile phase, giving rise to smaller van Deemter C terms and improved mass transfer, and the possibility of operating columns at high flow rates with reduced losses in efficiency for fast analysis [5]. Surprisingly good peak shapes can be obtained for some basic compounds. For example, efficiencies of around 100,000 plates/m with asymmetry factors (As) close to 1.0 were reported for basic drugs such as nortritpyline (pKa ∼10) using a 5-µm particle size bare silica HILIC phase. In comparison, such solutes often give rise to peak asymmetry in RP separations.

Figure 1.1. Yearly publications containing the terms “hydrophilic interaction chromatography” or “hydrophilic interaction liquid chromatography” or HILIC according to Thomson Web of Knowledge [1].

A separate advantage of HILIC is its compatibility with sample preparation methods using solid-phase extraction (SPE). Some such methods incorporate an elution step that uses a high concentration of an organic solvent, which gives rise to a potential injection solvent of the eluate that is stronger than typical RP mobile phases [2]. This mismatch in solvent strengths can give rise to peak broadening or splitting, necessitating evaporation of the SPE eluate and reconstitution in the mobile phase. SPE eluates with high organic solvent concentrations can be injected directly in HILIC, as they are weak solvents in this technique. The combination of different retention mechanisms in sample purification and analysis steps (HILIC/RP) can be advantageous in giving extra selectivity compared with an RP/RP procedure, where in some cases the SPE column may act merely as a sort of filter for the analytical column [6].