Methods in Molecular Biology TM Methods in Molecular Biology TM Edited by Gerald Gübitz Martin G. Schmid Chiral Separations Methods and Protocols VOLUME 243 Edited by Gerald Gübitz Martin G. Schmid Chiral Separations Methods and Protocols Chiral Separations M E T H O D S I N M O L E C U L A R B I O L O G Y TM Humana Press Totowa, New Jersey Chiral Separations Methods and Protocols Edited by Gerald Gübitz and Martin G. Schmid Institute of Pharmaceutical Chemistry and Pharmaceutical Technology, Karl-Franzens University, Graz, Austria © 2004 Humana Press Inc. 999 Riverview Drive, Suite 208 Totowa, New Jersey 07512 www.humanapress.com All rights reserved. No part of this book may be reproduced, stored in a retrieval system, or transmitted in any form or by any means, electronic, mechanical, photocopying, microfilming, recording, or otherwise without written permis- sion from the Publisher. 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Printed in the United States of America. 10 9 8 7 6 5 4 3 2 1 1-59259-648-7 (e-book) Library of Congress Cataloging in Publication Data Chiral separations : methods and protocols / edited by Gerald Gübitz and Martin G. Schmid. p. cm. (Methods in molecular biology ; 243) Includes bibliographical references and index. ISBN 1-58829-150-2 (alk. paper) ISSN 1064-3745 1. Enantiomers Separation Laboratory manuals. 2. Chirality. I. Gübitz, Gerald. II. Schmid, Martin G. III. Series. QP517.C57C46 2003 615'.19 dc21 200347771 Preface v Many compounds of biological and pharmacological interest are asym- metric and show optical activity. Approximately 40% of the drugs in use are known to be chiral and only about 25% are administered as pure enantiomers. It is well established that the pharmacological activity is mostly restricted to one of the enantiomers (eutomer). In several cases, unwanted side effects or even toxic effects may occur with the inactive enantiomer (distomer). Even if the side effects are not that drastic, the inactive enantiomer has to be metabo- lized, which represents an unnecessary burden for the organism. The adminis- tration of pure, pharmacologically active enantiomers is therefore of great importance. The ideal way to get to pure enantiomers would be by enantioselective synthesis. However, this approach is usually expensive and not often practicable. Usually, the racemates are obtained in a synthesis, and the separation of the enantiomers on a preparative scale is necessary. On the other hand, there is also a great demand for methods of enantiomer separation on an analytical scale for controlling synthesis, checking for racemization pro- cesses, controlling enantiomeric purity, and for pharmacokinetic studies. Con- ventional methods for enantiomer separation on a preparative scale are fractionated crystallization, the formation of diastereomeric pairs followed by repeated recrystallization, and enzymatic procedures. In recent years, chro- matographic methods such as gas chromatography and, especially, liquid chro- matography have attracted increasing interest for chiral separation, both on analytical and preparative scales. More recently, capillary electrophoresis and electrochromatography have also proven useful for chiral separation on an analytical scale. Chiral Separations: Methods and Protocols focuses on chromatographic and electroseparation techniques for chiral separation on an analytical scale. It is not the aim of this book to give a comprehensive overview of all applica- tions of chiral separation principles. Because there are several thousand pub- lications on this topic, this would require a series of books. For comprehensive overviews the reader is referred to specialized review articles. Chiral Separations: Methods and Protocols begins with an introduction to the different techniques, principles, and mechanisms of chiral separation, and includes a historical background (Chapter 1). Chapters 2–4 review some special techniques and include practical advice for users. The remainder of the book is devoted to articles describing typical procedures for enantiomer vi Preface separation by chromatographic and electromigration techniques applying dif- ferent chiral separation principles. These procedures may be of general char- acter, or are otherwise presented by means of applications to substance classes or special compounds. These chapters differ from conventional articles, because primary emphasis is set on giving reliable procedures for users. Spe- cial attention is given to important experimental data, and practical hints in the “Notes” section enable the reader to adapt these procedures to one’s sepa- ration problems. Forty-three authors from twenty-four research laboratories all over the world have contributed to Chiral Separations: Methods and Protocols. We want to express our thanks to all of our authors and coauthors for making their exper- tise and knowledge available to those who are not already versed in this area. This book should be helpful to biochemists, pharmaceutical chemists, clinical chemists, molecular biologists, and pharmacologists, both in research institutions and in industry. Gerald Gübitz Martin G. Schmid Contents Preface v Contributors xi 1 Chiral Separation Principles: An Introduction Gerald Gübitz and Martin G. Schmid 1 2 Separation of Enantiomers by Thin-Layer Chromatography: An Overview Kurt Günther, Peter Richter, and Klaus Möller 29 3 Cyclodextrin-Based Chiral Stationary Phases for Liquid Chromatography: A Twenty-Year Overview Clifford R. Mitchell and Daniel W. Armstrong 61 4 Enantiomeric Separations by HPLC Using Macrocyclic Glycopeptide-Based Chiral Stationary Phases: An Overview Tom Ling Xiao and Daniel W. Armstrong 113 5 Chiral Separation by HPLC Using Polysaccharide-Based Chiral Stationary Phases Chiyo Yamamoto and Yoshio Okamoto 173 6 Applications of Polysaccharide-Based Chiral Stationary Phases for Resolution of Different Compound Classes Hassan Y. Aboul-Enein and Imran Ali 183 7 Chiral Separation by HPLC With Pirkle-Type Chiral Stationary Phases Myung Ho Hyun and Yoon Jae Cho 197 8 Chiral Separation by HPLC Using the Ligand-Exchange Principle Vadim A. Davankov 207 9 Chiral Separations by HPLC Using Molecularly Imprinted Polymers Peter Spégel, Lars I. Andersson, and Staffan Nilsson 217 10 Indirect Enantioseparation by HPLC Using Chiral Benzofurazan-Bearing Reagents Toshimasa Toyo'oka 231 vii viii Contents 11 Separation of the Racemic Trans-Stilbene Oxide by Sub-/Supercritical Fluid Chromatography Leo Hsu, Genevieve Kennedy, and Gerald Terfloth 247 12 Chiral Separations Using Macrocyclic Antibiotics in Capillary Electrophoresis Timothy J. Ward and Colette M. Rabai 255 13 Enantioresolutions by Capillary Electrophoresis Using Glycopeptide Antibiotics Salvatore Fanali 265 14 Separation of Enantiomers by Capillary Electrophoresis Using Cyclodextrins Wioleta Maruszak, Martin G. Schmid, Gerald Gübitz, Elzbieta Ekiert, and Marek Trojanowicz 275 15 Chiral Separations by Capillary Electrophoresis Using Proteins as Chiral Selectors Jun Haginaka 291 16 Cellulases as Chiral Selectors in Capillary Electrophoresis Gunnar Johansson, Roland Isaksson, and Göran Pettersson 307 17 Use of Chiral Crown Ethers in Capillary Electrophoresis Martin G. Schmid and Gerald Gübitz 317 18 Chiral Separations by Capillary Electrophoresis Using Cinchona Alkaloid Derivatives as Chiral Counter-Ions Michael Lämmerhofer and Wolfgang Lindner 323 19 Chiral Separation by Capillary Electrophoresis Using Polysaccharides Hiroyuki Nishi 343 20 Chiral Micellar Electrokinetic Chromatography Koji Otsuka and Shigeru Terabe 355 21 Chiral Separation by Capillary Electrophoresis in Nonaqueous Medium Marja-Liisa Riekkola and Heli Sirén 365 22 Chiral Ligand-Exchange Capillary Electrophoresis and Capillary Electrochromatography Martin G. Schmid and Gerald Gübitz 375 23 Enantioseparation in Capillary Chromatography and Capillary Electrochromatography Using Polysaccharide-Type Chiral Stationary Phases Bezhan Chankvetadze 387 24 Chiral Separation by Capillary Electrochromatography Using Cyclodextrin Phases Dorothee Wistuba, Jingwu Kang, and Volker Schurig 401 25 Chiral Separations by Capillary Electrochromatography Using Molecularly Imprinted Polymers Peter Spégel, Jakob Nilsson, and Staffan Nilsson 411 Index 425 Contents ix xi Contributors HASSAN Y. ABOUL-ENEIN • Pharmaceutical Analysis Laboratory, Biological and Medical Research Department (MBC-03), King Faisal Specialist Hospital and Research Center, Riyadh, Saudi Arabia I MRAN ALI • Pharmaceutical Analysis Laboratory, Biological and Medical Research Department (MBC-03), King Faisal Specialist Hospital and Research Center, Riyadh, Saudi Arabia L ARS I. ANDERSSON • DMPK and Bioanalytical Chemistry, AstraZeneca Research and Development, Södertälje, Sweden D ANIEL W. ARMSTRONG • Department of Chemistry, Iowa State University, Ames, IA B EZHAN C HANKVETADZE • Molecular Recognition and Separation Science Laboratory, School of Chemistry, Tbilisi State University, Tbilisi, Georgia YOON JAE CHO • Department of Chemistry, Pusan National University, Pusan, South Korea V ADIM A. DAVANKOV • Institute of Organoelement Compounds (INEOS), Russian Academy of Sciences, Moscow, Russia E LZBIETA EKIERT • Department of Chemistry, Warsaw University, Warsaw, Poland S ALVATORE FANALI • Istituto di Metodologie Chimiche, C. N. R., Area della Ricerca di Roma, Monterotondo Scalo (Roma) Italy G ERALD GÜBITZ • Institute of Pharmaceutical Chemistry and Pharmaceutical Technology, Karl-Franzens University, Graz, Austria K URT G ÜNTHER • Degussa AG, Industriepark Wolfgang GmbH, Hanau, Germany JUN HAGINAKA • Faculty of Pharmaceutical Sciences, Mukogawa Women’s University, Nishinomiya, Japan L EO H SU • Research and Development, GlaxoSmithKline, King of Prussia, PA MYUNG HO HYUN • Department of Chemistry, Pusan National University, Pusan, South Korea R OLAND ISAKSSON • Department of Chemistry and Biomedical Sciences, University of Kalmar, Kalmar, Sweden G UNNAR JOHANSSON • Department of Biochemistry, Uppsala University, Uppsala, Sweden [...]... are heptakis-O-methyl-CD, heptakis (2,6-di-O-methyl)-CD, heptakis (2,3,6-triO-methyl)-CD, hydroxyethyl-CD, and hydroxypropyl-CD Since neutral CDs migrate with the same velocity as the electroosmotic flow (EOF), they cannot be used for neutral analytes Negatively charged CDs, such as sulfated CDs, sulfobutyl- and sulfoethyl-β-CD, carboxymethyl-β-CD, and succinyl-β-CD, were applied to the chiral separation... Used as Chiral Selectors Protein Trade name of CSP Manufacturer BSA Chiral BSA Resolvosil BSA-7 Resolvosil BSA-7PX Ultron ES-BSA Chiral- HSA Chiral HSA Chiral- AGP Shandon Nagel-Macherey Nagel-Macherey Shinwa Chemical Ind Chrom Tech AB Shandon Chrom Tech AB Ultron ES-OVM Bioptic AV-1 Shinwa Chemical Ind GL Sciences/Ansys Techn Chiral- CBH Chrom Tech AB Ultron ES-Pepsin Shinwa Chemical Ind HSA α1-Acid glycoprotein... 4 Gübitz and Schmid The formation of ion pairs using chiral counter ions such as (+)-S-10-camphor sulfonic acid (57,58), N-benzoylcarbonyl glycyl-L-proline (59), (-) 2,3,4,6di-O-isopropylidene-2-keto-L-gulonic acid (60), and quinine (59,61,62) was utilized for the HPLC separation of various basic and acidic drugs, respectively Also, with this principle, lateral binding forces have to support chiral recognition... between phenylalaninamide-based stationary and mobile phases Chirality 8, 452–461 84 Gübitz, G., Mihellyes, S., Kobinger, G., and Wutte, A (1994) New chemically bonded chiral ligand-exchange chromatographic stationary phases J Chromatogr A 666, 91–97 85 Wachsmann, M and Brückner, H (1998) Ligand-exchange chromatographic separation of DL-amino acids on aminopropylsilica-bonded chiral s-triazines Chromatographia... high-performance liquid-chromatography on a new chiral ligand-exchange phase J Chromatogr 592, 291–296 91 Wan, Q H., Shaw, P N., Davies, M C., and Barrett, D A (1997) Role of alkyl and aryl substituents in chiral ligand exchange chromatography of amino acids study using porous graphitic carbon coated with N-substituted-L-proline selectors J Chromatogr A 786, 249–257 92 Gil-Av, E., Tishbee, A., and. .. hydroxy acids (77), and thyroid hormones (78) Phases of this type have been commercialized by Serva, Heidelberg, Germany (Chiral= Si-L-Pro, L -Hypro, L-Val) and Daicel, Tokyo Japan (Chiralpak WH) Subsequently, a considerably high number of chiral LEC-phases has been published (79–85) Instead of chemically binding of ligands to silica gel, LEC-phases were also prepared by coating ligands with hydrophobic... of n-acylated 1-aryl-1-aminoalkanes J Org Chem 48, 5022–5026 47 Welch, C J (1994) Evolution of chiral stationary phase design in the Pirkle laboratories J Chromatogr A 666, 3–26 48 Hyun, M H and Min, C S (1998) Chiral recognition mechnism for the resolution of enantiomers on a highly effective HPLC chiral stationary phase derived from (R )-4 -hydroxyphenylglycine Chirality 10, 592–599 49 Lin, C.-E and. .. chiral selectors I Brush type chiral stationary phases for high-performance liquid chromatography based on cinchonan carbamates and their application as chiral anion exchangers J Chromatogr A 741, 33–48 55 Lämmerhofer, M and Lindner, W (1998) High-efficiency chiral separations of N-derivatized amino acids by packed-capillary electrochromatography with a quinine-based chiral anion-exchange type stationary... Pettersson, C and Karlsson, A (1992) Separation of enantiomeric amines and acids using chiral ion-pair chromatography on porous graphitic carbon Chirality 4, 323–332 60 Pettersson, C and Gioeli, C (1993) Chiral separation of amines using reversedphased ion-pair chromatography Chirality 5, 241–245 61 Pettersson, C and No, K (1983) Chiral resolution of carboxylic and sulfonic acids by ion-pair chromatography... review gives an overview of developments and applications of this technique (95) More recently, LE was also successfully applied in CEC Schmid et al (96) prepared an LE-continuous bed by in situ co-polymerization of methacrylamide and N-(2-hydroxy-3-allyloxypropyl)-L-4-hydroxyproline as a chiral selector in the presence of piperazine diacrylamide as a crosslinker and vinylsulfonic acid as a charge providing . Schmid Chiral Separations Methods and Protocols Chiral Separations M E T H O D S I N M O L E C U L A R B I O L O G Y TM Humana Press Totowa, New Jersey Chiral Separations Methods and Protocols Edited. 4 3 2 1 1-5 925 9-6 4 8-7 (e-book) Library of Congress Cataloging in Publication Data Chiral separations : methods and protocols / edited by Gerald Gübitz and Martin G. Schmid. p. cm. (Methods in. (109) and charged derivatives (110,111) were used. The most frequently used neutral CD derivatives are heptakis-O-methyl-CD, heptakis (2,6-di-O-methyl)-CD, heptakis (2,3,6-tri- O-methyl)-CD, hydroxyethyl-CD,