Chiral Separations and Stereochemical Elucidation: Fundamentals, Methods, and Applications Quezia Bezerra Cass
Visit to download the full and correct content document: https://ebookmass.com/product/chiral-separations-and-stereochemical-elucidation-fun damentals-methods-and-applications-quezia-bezerra-cass/
More products digital (pdf, epub, mobi) instant download maybe you interests ...
Electrochemical Methods: Fundamentals and Applications, 2nd Edition – Ebook PDF Version
https://ebookmass.com/product/electrochemical-methodsfundamentals-and-applications-2nd-edition-ebook-pdf-version/
Chiral Analysis: Advances in Spectroscopy, Chromatography and Emerging Methods P L Polavarapu
https://ebookmass.com/product/chiral-analysis-advances-inspectroscopy-chromatography-and-emerging-methods-p-l-polavarapu/
Molecular Diagnostics Fundamentals, Methods and Clinical Applications 2nd Edition, (Ebook PDF)
https://ebookmass.com/product/molecular-diagnostics-fundamentalsmethods-and-clinical-applications-2nd-edition-ebook-pdf/
Chiral Building Blocks in Asymmetric Synthesis: Synthesis and Applications Elzbieta Wojaczynska
https://ebookmass.com/product/chiral-building-blocks-inasymmetric-synthesis-synthesis-and-applications-elzbietawojaczynska/
Computational and Data-Driven Chemistry Using Artificial Intelligence: Fundamentals, Methods and Applications Takashiro Akitsu
https://ebookmass.com/product/computational-and-data-drivenchemistry-using-artificial-intelligence-fundamentals-methods-andapplications-takashiro-akitsu/
Fundamentals of Advanced Mathematics V3 (New Mathematical Methods, Systems and Applications) Henri Bourles
https://ebookmass.com/product/fundamentals-of-advancedmathematics-v3-new-mathematical-methods-systems-and-applicationshenri-bourles/
Two-Dimensional Materials for Nonlinear Optics: Fundamentals, Preparation Methods, and Applications
Qiang Wang
https://ebookmass.com/product/two-dimensional-materials-fornonlinear-optics-fundamentals-preparation-methods-andapplications-qiang-wang/
Nano-Optics: Fundamentals, Experimental Methods, and Applications (Micro and Nano Technologies) 1st Edition
Sabu Thomas (Editor)
https://ebookmass.com/product/nano-optics-fundamentalsexperimental-methods-and-applications-micro-and-nanotechnologies-1st-edition-sabu-thomas-editor/
Membrane-Based Separations in Metallurgy: Principles and Applications 1st Edition Lan Ying Jiang
https://ebookmass.com/product/membrane-based-separations-inmetallurgy-principles-and-applications-1st-edition-lan-yingjiang/
Chiral Separations and Stereochemical Elucidation
Chiral Separations and Stereochemical Elucidation
Fundamentals,
Methods, and Applications
Edited by
Quezia Bezerra Cass
Universidade Federal de São Carlos
São Carlos, SP, Brazil
Maria Elizabeth Tiritan
Universidade do Porto
Porto, Portugal
João Marcos Batista Junior
Universidade Federal de São Paulo
São José dos Campos, SP, Brazil
Juliana Cristina Barreiro
Universidade de São Paulo
São Carlos, SP, Brazil
Copyright © 2023 by John Wiley & Sons, Inc. All rights reserved.
Published by John Wiley & Sons, Inc., Hoboken, New Jersey. Published simultaneously in Canada.
No part of this publication may be reproduced, stored in a retrieval system, or transmitted in any form or by any means, electronic, mechanical, photocopying, recording, scanning, or otherwise, except as permitted under Section 107 or 108 of the 1976 United States Copyright Act, without either the prior written permission of the Publisher, or authorization through payment of the appropriate per-copy fee to the Copyright Clearance Center, Inc., 222 Rosewood Drive, Danvers, MA 01923, (978) 750-8400, fax (978) 750-4470, or on the web at www.copyright.com. Requests to the Publisher for permission should be addressed to the Permissions Department, John Wiley & Sons, Inc., 111 River Street, Hoboken, NJ 07030, (201) 748-6011, fax (201) 748-6008, or online at http://www.wiley.com/go/permission.
Trademarks: Wiley and the Wiley logo are trademarks or registered trademarks of John Wiley & Sons, Inc. and/or its affiliates in the United States and other countries and may not be used without written permission. All other trademarks are the property of their respective owners. John Wiley & Sons, Inc. is not associated with any product or vendor mentioned in this book.
Limit of Liability/Disclaimer of Warranty:
While the publisher and author have used their best efforts in preparing this book, they make no representations or warranties with respect to the accuracy or completeness of the contents of this book and specifically disclaim any implied warranties of merchantability or fitness for a particular purpose. No warranty may be created or extended by sales representatives or written sales materials. The advice and strategies contained herein may not be suitable for your situation. You should consult with a professional where appropriate. Further, readers should be aware that websites listed in this work may have changed or disappeared between when this work was written and when it is read. Neither the publisher nor authors shall be liable for any loss of profit or any other commercial damages, including but not limited to special, incidental, consequential, or other damages.
For general information on our other products and services or for technical support, please contact our Customer Care Department within the United States at (800) 762-2974, outside the United States at (317) 572-3993 or fax (317) 572-4002.
Wiley also publishes its books in a variety of electronic formats. Some content that appears in print may not be available in electronic formats. For more information about Wiley products, visit our web site at www.wiley.com.
Library of Congress Cataloging-in-Publication Data applied for: ISBN: 9781119802259 (HB); ePDF: 9781119802273; epub: 9781119802266
Cover Design: Wiley
Cover Image: © kcarper/500px/Getty Images
Set in 9.5/12.5pt STIXTwoText by Straive, Pondicherry, India
Contents
List of Contributors xv
Preface xix
Part I Fundamentals of Chiral Separation 1
1 Chiral Separation by LC 3
Juliana Cristina Barreiro and Quezia Bezerra Cass
1.1 Introduction 3
1.2 Workflow for LC Chiral Method Development 7
1.3 New Column Technologies 9
1.4 Selected Examples of Fast Separation 12
1.5 Chiral 2D-LC 14
1.5.1 LC–LC and mLC–LC 14
1.5.2 LC × LC and sLC × LC 17
1.6 Future and Perspectives 19
References 20
2 Chiral Separation by GC 27
Oliver Trapp
2.1 Introduction 27
2.2 Chiral Recognition in Gas Chromatography 29
2.2.1 Chiral Recognition by Hydrogen Bonding 31
2.2.2 Chiral Recognition Using Chiral Metal Complexes 31
2.2.3 Chiral Recognition by Host–Guest Interactions 31
2.3 Preparation of Fused-Silica Capillaries for GC with CSPs 33
2.4 Application of CSPs in Chiral Gas Chromatography 34
2.4.1 CSPs with Diamide Selectors 34
2.4.1.1 Chirasil-Val 34
2.4.2 CSPs with CD Selectors 35
2.4.2.1 Heptakis(2,3,6-tri-O-Methyl)-β- Cyclodextrin (Permethyl-β- Cyclodextrin) 38
2.4.2.2 Heptakis(2,3,6-tri-O-Methyl)-β- Cyclodextrin Immobilized to Hydrido Dimethyl Polysiloxane (Chirasil-β-Dex) 39
2.4.2.3 Heptakis(2,6-di-O-Methyl-3-O-Pentyl)-β- Cyclodextrin 43
2.4.2.4 Hexakis-(2,3,6-tri-O-Pentyl)-α- Cyclodextrin 47
2.4.2.5 Heptakis(2,3,6-tri-O-Pentyl)-β- Cyclodextrin 48
2.4.2.6 Hexakis-(3-O-Acetyl-2,6-di-O-Pentyl)-α- Cyclodextrin 51
2.4.2.7 Heptakis(3-O-Acetyl-2,6-di-O-Pentyl)-β- Cyclodextrin 51
2.4.2.8 Octakis(3-O-Butyryl-2,6-di-O-Pentyl)-γ- Cyclodextrin 53
2.4.2.9 Hexakis/Heptakis/Octakis(2,6-di-O-Alkyl-3-O-Trifluoroacetyl)α/β/γ- Cyclodextrins 57
2.4.2.10 Heptakis(2,3-di-O-Acetyl-6-O-tert-Butyldimethylsilyl)-βCyclodextrin (DIAC-6-TBDMS-β- CD) 58
2.4.2.11 Heptakis(2,3-di-O-Methyl-6-O-tert-Butyldimethylsilyl)-βCyclodextrin (DIME-6-TBDMS-β- CD) 58
2.4.3 Cyclofructans 62
2.4.4 CSPs with Metal Complexes 65
2.5 Conclusion 69 References 69
3 Chiral Separation by Supercritical Fluid Chromatography 85 Emmanuelle Lipka
3.1 Introduction 85
3.2 Characteristics and Properties of Supercritical Fluids 87
3.3 Development of a Chiral SFC Method 89
3.3.1 Chiral Stationary Phases 89
3.3.2 Mobile Phases 91
3.3.2.1 Mobile Phase: Type of Co-solvent Used 93
3.3.2.2 Mobile Phase: Percentage of Co-solvent Used 94
3.3.2.3 Mobile Phase: Use of Additives 94
3.4 Operating Parameters 94
3.4.1 Effect of the Flow Rate 95
3.4.2 Effect of the Outlet Pressure (Back-pressure) 95
3.4.2.1 Effect of Pressure When the Mobile Phase is a Gas-Like Fluid 96
3.4.2.2 Effect of Pressure When the Mobile Phase is a Liquid-Like Fluid 97
3.4.3 Effect of Temperature 97
3.4.3.1 Effect of Temperature When the Mobile Phase is a Gas-Like Fluid 98
3.4.3.2 Effect of Temperature When the Mobile Phase is a Liquid-Like Fluid 98
3.5 Detection 99
3.6 Scale-Up to Preparative Separation 99
3.7 Conclusion 100 References 101
4 Chiral Separation by Capillary Electrophoresis and Capillary Electrophoresis–Mass Spectrometry: Fundamentals, Recent Developments, and Applications 103 Charles Clark, Govert W. Somsen, and Isabelle Kohler
4.1 Introduction 103
4.2 Principles of Chiral CE 105
4.2.1 Electrophoretic Mobility 105
4.2.2 CE Separation Efficiency 106
4.2.3 Chiral Resolution in CE 107
4.2.4 Chiral Micellar Electrokinetic Chromatography and Capillary Electrochromatography 109
4.3 Short History of Chiral CE Modes 111
4.3.1 Chiral CE 111
4.3.2 Chiral MEKC and Chiral CEC 111
4.4 State of the Art and Recent Developments 112
4.4.1 Common Chiral Selectors 112
4.4.2 Ionic Liquids as Chiral Selectors 117
4.4.3 Nanoparticles as Chiral Selector Carriers 117
4.4.4 Microfluidic Chiral CE 118
4.5 Applications of Chiral CE 119
4.5.1 Pharmaceutical Analysis 119
4.5.2 Food Analysis 120
4.5.3 Environmental Analysis 121
4.5.4 Bioanalysis 123
4.5.5 Forensic Analysis 126
4.6 Chiral CE-MS: Strategies and Challenges 126
4.6.1 Hyphenation Approaches 129
4.6.1.1 Sheath–Liquid and Sheathless CE-MS Interfacing 129
4.6.1.2 Partial-Filling Techniques 130
4.6.1.3 Counter-Migration Techniques 131
4.6.2 Chiral MEKC-MS 132
4.6.3 Chiral CEC-MS 133
4.7 Conclusions and Perspectives 135 References 135
5 Chiral Separations at Semi and Preparative Scale 143 Larry Miller
5.1 Introduction 143
5.2 Selection of Operating Conditions 145
5.3 Batch HPLC Purification 146
5.3.1 Analytical Method Development for Preparative Separations 146
5.3.2 Batch HPLC Examples 148
5.3.2.1 Batch HPLC Example 1 148
5.3.2.2 Batch HPLC Example 2 149
5.4 Steady-State Recycle Introduction 151
5.4.1 SSR Example 1 153
5.5 Simulated Moving Bed Chromatography – Introduction 154
5.5.1 SMB Examples for R&D and Separation of Compound 2 156
5.5.2 Development of a Manufacturing SMB Process (Compound 1) 158
5.5.3 Cost for SMB Processes 160
5.6 Introduction to Supercritical Fluid Chromatography 161
5.6.1 Analytical Method Development for Scale-up to Preparative SFC 162
5.6.2 Preparative SFC Example 1 163
5.6.3 Preparative SFC Example 2 163
5.7 Options for Increasing Purification Productivity 165
5.7.1 Closed-Loop Recycling 165
5.7.2 Stacked Injections 166
5.7.3 Choosing the Best Synthetic Intermediate for Separation 167
5.7.3.1 Choosing Synthetic Step for Separation – HPLC/SMB Example 168
5.7.3.2 Choosing Synthetic Step for Separation – SFC Example 169
5.7.4 Use of Non- Commercialized CSP 170
5.7.5 Immobilized CSP for Preparative Resolution 173
5.7.5.1 Processing of Low Solubility Racemate 173
5.7.5.2 Preparative Resolution of EMD 53986 174
5.8 Choosing a Technique for Preparative Enantioseparation 176
5.9 Conclusion 178 References 179
Part II
Chiral Selectors 187
6 Polysaccharides 189 Weston Umstead, Takafumi Onishi, and Pilar Franco
6.1 Introduction 189
6.2 The Early Years 190
6.3 Polysaccharide Chiral Separation Mechanism 193
6.4 Coated Chiral Stationary Phases 197
6.5 Immobilized Chiral Stationary Phases 201
6.6 Applications of Polysaccharide-Derived CSPs 208
6.6.1 Analytical Applications 210
6.6.1.1 Pharmaceuticals 211
6.6.1.2 Agrochemicals 218
6.6.1.3 Food Analysis 219
6.6.2 Preparative Applications 220
6.7 Summation 224
References 224
7 Macrocyclic Antibiotics and Cyclofructans 247
Saba Aslani, Alain Berthod, and Daniel W. Armstrong
7.1 Introduction 247
7.2 Macrocyclic Glycopeptides Physicochemical Properties 248
7.3 Using the Chiral Macrocyclic Glycopeptides Stationary Phases 253
7.3.1 Mobile Phases and Chromatographic Modes 253
7.3.2 Chromatographic Enantioseparations 254
7.3.2.1 Amino Acids and Peptides 254
7.3.2.2 Chiral Compounds 257
7.3.2.3 Particle Structure 257
7.4 Using and Protecting Macrocyclic Glycopeptide Chiral Columns 260
7.4.1 Operating Conditions 260
7.4.2 Storage 261
7.5 Cyclofructans 261
7.5.1 Cyclofructan Structure and Properties 261
7.5.2 Chiral Separations with Cyclofructan-Based Stationary Phases 264
7.5.3 Cyclofructan Stationary Phases Used in the HILIC Mode 264
7.5.4 Cyclofructan Stationary Phases Used in Supercritical Fluid Chromatography 266
7.6 Conclusions 267
References 268
8 Cyclodextrins 273
Gerhard K. E. Scriba, Mari-Luiza Konjaria, and Sulaiman Krait
8.1 Introduction 273
8.2 Structure and Properties 274
8.3 Cyclodextrin Complexes 279
8.4 Application in Separation Science 288
Contents x
8.4.1 Gas Chromatography 288
8.4.1.1 Types of Cyclodextrins 289
8.4.1.2 Types of Columns 289
8.4.1.3 Separation Mechanisms 291
8.4.1.4 Applications 293
8.4.2 Thin-Layer Chromatography 294
8.4.3 High-Performance Liquid Chromatography 294
8.4.3.1 Types of Columns 295
8.4.3.2 Types of Cyclodextrins 297
8.4.3.3 Separation Mechanisms 298
8.4.3.4 Applications 300
8.4.4 Supercritical Fluid Chromatography 300
8.4.5 Capillary Electromigration Techniques 301
8.4.5.1 Types of Cyclodextrins 301
8.4.5.2 Separation Mechanisms 302
8.4.5.3 Migration Modes and Enantiomer Migration Order Using CDs as Selectors 304
8.4.5.4 Applications 310
8.4.6 Membrane Technologies 312
8.5 Miscellaneous Applications 314
8.6 Conclusions and Outlook 315 References 315
9 Pirkle Type 325
Maria Elizabeth Tiritan, Madalena Pinto, and Carla Fernandes
9.1 Introduction 325
9.2 CSPs Developed by Pirkle’s Group: Chronological Evolution 327
9.3 Pirkle-Type CSPs Developed by Other Research Groups 334
9.4 Example of Applications in Analytical and Preparative Scales 340
9.4.1 Analytical Applications 341
9.4.2 Preparative Applications 349
9.5 Conclusions and Perspectives 349 References 350
10 Proteins 363
Jun Haginaka
10.1 Introduction 363
10.2 Preparation of Protein- and Glycoprotein-Based Chiral Stationary Phases 364
10.3 Types of Protein- and Glycoprotein-Based Chiral Stationary Phases 368
10.3.1 Proteins 368
10.3.1.1 Bovine Serum Albumin 368
10.3.1.2 Human Serum Albumin 370
10.3.1.3 Trypsin and α- Chymotrypsin 372
10.3.1.4 Lysozyme and Pepsin 372
10.3.1.5 Fatty Acid-Binding Protein 373
10.3.1.6 Penicillin G Acylase 375
10.3.1.7 Streptavidin 375
10.3.1.8 Lipase 376
10.3.2 Glycoproteins 376
10.3.2.1 Human α1-Acid Glycoprotein 376
10.3.2.2 Chicken Ovomucoid 377
10.3.2.3 Chicken α1-Acid Glycoprotein 378
10.3.2.4 Avidin 380
10.3.2.5 Riboflavin-Binding Protein and Ovotransferrin 380
10.3.2.6 Cellobiohydrolase 381
10.3.2.7 Glucoamylase 383
10.3.2.8 Antibody (Immunoglobulin G) 385
10.3.2.9 Nicotinic Acetylcholine Receptor and Human Liver Organic Cation Transporter 387
10.4 Chiral Recognition Mechanisms on Proteinand Glycoprotein-Based Chiral Stationary Phases 387
10.4.1 Human Serum Albumin 387
10.4.2 Penicillin G Acylase 389
10.4.3 Human α1-Acid Glycoprotein 390
10.4.4 Turkey Ovomucoid 392
10.4.5 Chicken α1-Acid Glycoprotein 393
10.4.6 Cellobiohydrolase 395
10.4.7 Antibody 396
10.4.8 Nicotinic Acetylcholine Receptor and Human Liver Organic Cation Transporter 400
10.5 Conclusions 401 References 402
11 Chiral Stationary Phases Derived from Cinchona Alkaloids 415 Michael Lämmerhofer and Wolfgang Lindner
11.1 Introduction 415
11.2 Cinchona Alkaloid-Derived Chiral Stationary Phases 416
11.3 Chiral Recognition 420
11.4 Chromatographic Retention Mechanisms 424
11.4.1 Multimodal Applicability 424
11.4.2 Surface Charge of Cinchonan-Based CSPs 424
11.4.3 Retention Mechanisms and Models, and Method Development on Chiral WAX CSPs 427
11.4.4 Retention Mechanisms and Method Development on ZWIX CSPs 430
11.5 Structural Variants of Cinchona Alkaloid CSPs and Immobilization Chemistries 436
11.6 Cinchonan-Based UHPLC Column Technologies 442
11.7 Applications 446
11.7.1 Pharmaceutical and Biotechnological Applications 446
11.7.2 Biomedical Applications 453
11.8 Conclusions 460 References 460
Part III Methods for Stereochemical Elucidation 473
12 X-Ray Crystallography for Stereochemical Elucidation 475 Ademir F. Morel and Robert A. Burrow
12.1 Introduction 475
12.2 Absolute Structure and Absolute Configuration 476
12.3 Best Practices 482
12.4 Structure Validation 486
12.5 The Absolute Configuration of (+)-Lanatine A 486
12.6 The Absolute Configuration of the Diacetylated Form of Acrenol and the Acetylated Form of Humirianthol 488
12.7 The Absolute Configuration of Ester Form of Clemateol 491
12.8 Relative Configurations of Waltherione A, Waltherione B, and Vanessine 492
12.9 The Absolute Configuration of Condaline A 493
12.10 CSD Deposit Numbers 496
12.11 Conclusions and Future Directions 498 References 498
13 NMR for Stereochemical Elucidation 505 Xiaolu Li, Xiaoliang Yang, and Han Sun
13.1 Conventional NMR Methods for Stereochemical Elucidation 505
13.1.1 Determination of the Planar Structure Using 1D 1H, 13C NMR (DEPT), 2D HSQC, COSY, TOCSY, HMBC 506
13.1.2 Determination of Relative Configuration Using J- Couplings and NOEs/ROEs 507
13.1.2.1 Scalar Coupling 507
13.1.2.2 NOE/ROE 510
13.1.2.3 Examples of Stereochemical Elucidation Using J- Couplings and NOEs/ROEs 510
13.2 Determination of the Relative Configuration Using Anisotropic NMR-Based Methods 516
13.2.1 Basic Principles of Anisotropic NMR Parameters 517
13.2.2 Alignment Media 518
13.2.2.1 Preparation of Anisotropic Sample with PMMA Gel 520
13.2.2.2 Preparation of Anisotropic Sample with AAKLVFF 521
13.2.3 Acquisition of the Anisotropic NMR Data 522
13.2.4 Computational Approaches for Analyzing Anisotropic NMR Data 525
13.2.5 Successful Examples of Determination of Relative Configuration of Challenging Molecules Using Anisotropic NMR 528
13.3 Determination of the Relative Configuration Using DP4 Probability and CASE-3D 529
13.4 Determination of the Absolute Configuration Using a Combination of NMR Spectroscopy and Chiroptical Spectroscopy 533
13.5 Determination of the Absolute Configuration Using NMR Alone 534
13.5.1 Mosher Ester Analysis 535
13.5.2 Other Chiral Derivatizing Agents 536
13.6 Future Perspective 536 References 537
14 Absolute Configuration from Chiroptical Spectroscopy 551 Fernando Martins dos Santos Junior and João Marcos Batista Junior
14.1 Introduction 551
14.2 Chiroptical Methods 554
14.2.1 Optical Rotation and Optical Rotatory Dispersion 554
14.2.1.1 Instrumentation 556
14.2.1.2 Measurements 557
14.2.2 Electronic Circular Dichroism 558
14.2.2.1 Instrumentation 560
14.2.2.2 Measurements 561
14.2.3 Vibrational Circular Dichroism and Raman Optical Activity 561
14.2.3.1 Instrumentation 563
14.2.3.2 Measurements 565
14.2.4 Simulation of Chiroptical Properties 567
14.2.4.1 Common Theoretical Steps 568
14.2.4.2 OR and ORD Simulations 570
14.2.4.3 ECD Simulations 572
14.2.4.4 VCD and ROA Simulations 573
14.2.5 Examples of Application 575
14.2.5.1 OR 575
14.2.5.2 ORD 577
14.2.5.3 ECD 578
14.2.5.4 VCD 579
14.2.5.5 ROA 581
14.2.5.6 Association of Different Chiroptical Methods 582
14.3 Concluding Remarks 585 References 586
Index 593
List of Contributors
Daniel W. Armstrong
Department of Chemistry and Biochemistry
University of Texas Arlington, TX, USA
Saba Aslani
Department of Chemistry and Biochemistry
University of Texas Arlington, TX, USA
Juliana Cristina Barreiro
Instituto de Química de São Carlos
Universidade de São Paulo
São Carlos, SP, Brazil
João Marcos Batista Junior
Instituto de Ciência e Tecnologia
Universidade Federal de São Paulo
São José dos Campos, SP, Brazil
Alain Berthod
Institute of Analytical Sciences
University of Lyon 1
CNRS, Villeurbanne, France
Quezia Bezerra Cass
Departamento de Química
Universidade Federal de São Carlos
São Carlos, SP, Brazil
Robert A. Burrow
Departamento de Química
Universidade Federal de Santa Maria RS, Brazil
Charles Clark
Leiden Academic Centre for Drug Research, Metabolomics and Analytics Centre, Leiden University Leiden, the Netherlands
Fernando Martins dos Santos Junior Instituto de Química Universidade Federal Fluminense Niterói, RJ, Brazil
Carla Fernandes
Laboratório de Química Orgânica e Farmacêutica
Departamento de Ciências Químicas Faculdade de Farmácia da Universidade do Porto Porto, Portugal and
Centro Interdisciplinar de Investigação Marinha e Ambiental (CIIMAR) Matosinhos, Portugal
Pilar Franco
Chiral Technologies Europe Illkirch Cedex, France
List of Contributors
Jun Haginaka Institute for Biosciences
Mukogawa Women’s University
Koshien Kyuban-cho Nishinomiya, Japan
Isabelle Kohler Division of BioAnalytical Chemistry
Amsterdam Institute of Molecular Life Sciences (AIMMS)
Vrije Universiteit Amsterdam Amsterdam, the Netherlands and Center for Analytical Sciences
Amsterdam
Amsterdam, the Netherlands
Mari-Luiza Konjaria
Department of Pharmaceutical/ Medicinal Chemistry
Friedrich Schiller University Jena Jena, Germany
Sulaiman Krait Department of Pharmaceutical/ Medicinal Chemistry
Friedrich Schiller University Jena Jena, Germany
Michael Lämmerhofer Institute of Pharmaceutical Sciences
University of Tübingen Tübingen, Germany
Xiaolu Li
Group of Structural Chemistry and Computational Biophysics
Leibniz-Forschungsinsitut für Molekulare Pharmakologie Berlin, Germany
Wolfgang Lindner Institute of Analytical Chemistry University of Vienna Vienna, Austria
Emmanuelle Lipka Laboratoire de Chimie Analytique –UFR3S-Faculté de Pharmacie de Lille, Inserm U1167, Université de Lille, Lille Cedex, France
Larry Miller Amgen Research
One Amgen Center Drive Thousand Oaks, CA, USA
Ademir F. Morel
Departamento de Química
Universidade Federal de Santa Maria RS, Brazil
Takafumi Onishi Daicel Corporation CPI Company Myoko-shi, Niigata, Japan
Madalena Pinto
Laboratório de Química Orgânica e Farmacêutica
Departamento de Ciências Químicas
Faculdade de Farmácia da Universidade do Porto Porto, Portugal and
Centro Interdisciplinar de Investigação
Marinha e Ambiental (CIIMAR) Matosinhos, Portugal
Gerhard K. E. Scriba
Department of Pharmaceutical/ Medicinal Chemistry
Friedrich Schiller University Jena Jena, Germany
Govert W. Somsen
Division of BioAnalytical Chemistry
Amsterdam Institute of Molecular Life Sciences (AIMMS)
Vrije Universiteit Amsterdam
Amsterdam, the Netherlands and
Center for Analytical Sciences
Amsterdam
Amsterdam, the Netherlands
Han Sun
Group of Structural Chemistry and Computational Biophysics
Leibniz-Forschungsinsitut für Molekulare Pharmakologie Berlin, Germany and Institute of Chemistry
Technical University of Berlin Berlin, Germany
List of Contributors
Maria Elizabeth Tiritan
Laboratório de Química Orgânica e Farmacêutica
Departamento de Ciências Químicas
Faculdade de Farmácia da Universidade do Porto Porto, Portugal and Centro Interdisciplinar de Investigação Marinha e Ambiental (CIIMAR)
Matosinhos, Portugal
Oliver Trapp Department of Chemistry
Ludwig-MaximiliansUniversity Munich Munich, Germany
Weston Umstead Chiral Technologies Inc. West Chester, PA, USA
Xiaoliang Yang
State Key Laboratory of Coordination Chemistry and Jiangsu Key Laboratory of Advanced Organic Materials School of Chemistry and Chemical Engineering, Nanjing University Nanjing, Jiangsu, China
Preface
The enchantment of reading about the first developments on chiral selectors for enantioseparation persists up to now. The papers from Hesse and Hagel [1], Pirkle [2, 3], Allenmark [4, 5], Okamoto [6], Armstrong [7] back in the 1970s–1980s, to cite a few, marked the vertiginous development of chiral stationary phases (CSPs).
Pirkle’s paper [8] about ionically bonded CSPs is a milestone paper. It pointed to the need of stablishing a relationship between absolute configuration (AC) and elution order providing a rationale for chiral recognition. This publication came alongside with the 3,5-dinitrobenzoyl phenylglycine CSP, the first commercial chiral column for liquid chromatography (LC) [9]. Ariens’ crusade [10] to raise awareness about the implication of stereochemistry in pharmacokinetics studies and, specially, in drug development programs and in clinical practice gave the tone for the development of appropriate chiral drug analysis procedures. The chiral analysis of drugs in biological fluids was, then, frenetically pursued in the 1980s and resulted in the FDA’s regulations on pharmaceutical development of single enantiomers and racemates [11]. The approval of a new chiral drug by regulatory agencies requires a complete dossier of the pharmacology and pharmacokinetic profiles of the single enantiomers and their mixtures. Methods to monitor the biological effects of a chiral drug are required since pre-clinical level with a few milligrams of racemates and single enantiomers moving to kilogram quantities at clinical trials. At each stage, it is important to know which is the most appropriate procedure either to the analysis or production of enantiomers. Such knowledge is thus considered a very important asset [12].
The advent of direct enantioseparation by chromatography and related techniques has just completed half a century, and it is nowadays used as routine in research and industry laboratories around the world. This book covers the main features of chiral separation by LC, gas chromatography
(GC), capillary electrophoresis (CE), and supercritical fluid chromatography (SFC). Chiral separation at semi- and preparative scales are also included.
With the five fundamental chapters (1–5) covering LC, GC, CE, SFC, and preparative chromatography, the book encompasses the main advances of each technique and discusses the application of chiral separation in different areas of science, such as enantioselective synthesis, chiral drug designing and development, bio, forensic, and environmental markers, chiral materials, quality control in pharmaceuticals, food, and fragrances. Chiral separations in metabolomics and lipidomics to disclose the enantiomeric signature in various biological processes is another topic that has been positively impacted by the advance in chiral selector technologies alongside with the appropriate analytical platforms. Application examples can be found throughout the book. The role of 2D-LC in chiral separation method development and applications is discussed in Chapter 1 as well as in other chapters of the book.
The quality of the separation in CE, GC, LC, and SFC is dictated by the chiral selector, and with so many interconnected interactions playing a role in chiral discrimination, it is expected that there will never be either a universal chiral selector or CSP.
The advances in chiral selectors and their use for all these separation technologies are thoroughly discussed from Chapters 6 to 11, including new CSP for LC and/or SFC with high-throughput and ultra-high-efficiency capabilities. These chapters portray the most used chiral selectors including their designing, mechanism, and applications for a variety of fields with practical examples. The impact on the chiral recognition mechanism caused by the elution mode in LC is also reviewed as well as the best chiral selectors either for SFC or for preparative separation.
Following chiral separation, the next challenge is the determination of molecular stereochemistry. Methods for stereochemical elucidation are widely used across biochemistry, chemistry, biology, and physics, but there is a clear need for streamlining their application for characterization of small chiral organic molecules.
The 3D structure characterization of a given chiral molecule is of prime importance, and, although not trivial, it is routinely required in many research areas, especially in drug discovery programs. For chiral molecules, prior to setting up several biological experiments, it is indispensable to unambiguously determine not only their stereoisomeric composition but also their AC [13, 14].
This approach ensures that a correct structure-activity relationship is established. The AC assignment of a given molecule is usually carried out by different methods in which X-ray crystallography is considered the gold
standard approach. Nuclear magnetic resonance (NMR) is routinely used for establishing relative configurations, while the AC can be determined either by diastereomeric derivatizations or by a combination of anisotropic NMR and chiroptical spectroscopy. Chiroptical methods, mainly associated with quantum-mechanical calculations, have proved to be an excellent choice to assign AC especially for compounds in which a well-defined single crystal is not available.
In this regard, Chapters 12–14 cover these three main stereochemical elucidation methods. Information on their theoretical backgrounds, advantages, and limitations, as well as examples of application are provided. With that, we do hope many research projects dealing with small organic molecules will benefit from a streamlined approach to both enantiomeric separation and AC determination.
By Quezia Bezerra Cass, Maria Elizabeth Tiritan, João Marcos Batista Junior, and Juliana Cristina Barreiro, São Carlos, August 2022
References
1 Hesse, G. and Hagel, R. (1973). Eine vollständige Recemattennung durch eluitons-chromagographie an cellulose-tri-acetat. Chromatographia 6: 277–280. https://doi.org/10.1007/BF02282825.
2 Pirkle, W.H. and Sikkenga, D.L. (1976). Resolution of optical isomers by liquid chromatography. J. Chromatogr. A 123: 400–404. https://doi.org/ 10.1016/S0021-9673(00)82210-4.
3 Pirkle, W.H., House, D.W., and Finn, J.M. (1980). Broad spectrum resolution of optical isomers using chiral high-performance liquid chromatographic bonded phases. J. Chromatogr. A 192: 143–158. https://doi.org/10.1016/ S0021-9673(00)81849-X.
4 Allenmark, S. and Bomgren, B. (1982). Direct liquid chromatographic separation of enantiomers on immobilized protein stationary phases. J. Chromatogr. A 252: 297–300. https://doi.org/10.1016/ S0021-9673(01)88421-1.
5 Allenmark, S., Bomgren, B., and Borén, H. (1983). Direct liquid chromatographic separation of enantiomers on immobilized protein stationary phases. J. Chromatogr. A 264: 63–68. https://doi.org/10.1016/ S0021-9673(01)95006-X.
6 Okamoto, Y., Kawashima, M., and Hatada, K. (1984). Chromatographic resolution. 7. Useful chiral packing materials for high-performance liquid
chromatographic resolution of enantiomers: phenylcarbamates of polysaccharides coated on silica gel. J. Am. Chem. Soc. 106: 5357–5359. https://doi.org/10.1021/ja00330a057.
7 Armstrong, D.W. and DeMond, W. (1984). Cyclodextrin bonded phases for the liquid chromatographic separation of optical, geometrical, and structural isomers. J. Chromatogr. Sci. 22: 411–415. https://doi.org/10.1093/ chromsci/22.9.411.
8 Pirkle, W.H., Finn, J.M., Schreiner, J.L., and Hamper, B.C. (1981). A widely useful chiral stationary phase for the high-performance liquid chromatography separation of enantiomers. J. Am. Chem. Soc. 103: 3964–3966. https://doi.org/10.1021/ja00403a076.
9 Pirkle, W.H., Myung, H.H., and Bank, B. (1984). A rational approach to the design of highly-effective chiral stationary phases. J. Chromatogr. A 316: 585–604. https://doi.org/10.1016/S0021-9673(00)96185-5.
10 Ariëns, E.J. (1984). Stereochemistry, a basis for sophisticated nonsense in pharmacokinetics and clinical pharmacology. Eur. J. Clin. Pharmacol. 26: 663–668. https://doi.org/10.1007/BF00541922.
11 (1992). FDA’S policy statement for the development of new stereoisomeric drugs. Chirality 4: 338–340. https://doi.org/10.1002/chir.530040513.
12 Tarafder, A. and Miller, L. (2021). Chiral chromatography method screening strategies: past, present and future. J. Chromatogr. A 1638: 461878. https://doi.org/10.1016/j.chroma.2021.461878.
13 Bogaerts, J., Aerts, R., Vermeyen, T. et al. (2021). Tackling stereochemistry in drug molecules with vibrational optical activity. Pharmaceuticals 14: 877. https://doi.org/10.3390/ph14090877.
14 Batista, A.N.L., dos Santos, F.M., Batista, J., and Cass, Q. (2018). Enantiomeric mixtures in natural product chemistry: separation and absolute configuration assignment. Molecules 23, 492: https://doi.org/ 10.3390/molecules23020492.
Part I
Fundamentals of Chiral Separation
1
Chiral Separation by LC
Juliana Cristina Barreiro1 and Quezia Bezerra Cass2
1Instituto de Química de São Carlos, Universidade de São Paulo, São Carlos, SP, Brazil
2Departamento de Química, Universidade Federal de São Carlos, São Carlos, SP, Brazil
1.1 Introduction
Liquid chromatography (LC) plays a central role in enantioseparation as it is used for analytical and preparative purposes.
Indirect chiral separation is still important in diverse application fields, mainly in metabolomic and lipidomics non-target analysis as well as in forensics [1–3]. For that, the formed diastereomeric mixtures are separated by achiral columns, mostly under reverse elution mode. Impurities of derivatization reagents that can lead to inaccuracies is one of the main drawbacks of the indirect approach. In the field of non-target metabolomics, several approaches have been described for derivatization of OH/ NH2 moiety- containing metabolites. In this regard, the use of diacetyl-tartaric anhydride (DATAN) has demonstrated its utility for identifying the enantiomers of hydroxycarboxylic acids (HAs) and amino acids (AAs) through the reversal in the elution order of the diastereomers formed using either (RR)-DATAN or (SS)-DATAN. Since the order of elution does not change for achiral metabolites, this approach has been used also to differentiate achiral metabolites from the chiral ones [4] (Scheme 1.1).
Direct chiral separation in LC can be achieved either by addition of a chiral selector to the mobile phase or by using chiral stationary phases (CSPs); herein we will discuss only the latter mode.
Chiral Separations and Stereochemical Elucidation: Fundamentals, Methods, and Applications, First Edition. Edited by Quezia Bezerra Cass, Maria Elizabeth Tiritan, João Marcos Batista Junior, and Juliana Cristina Barreiro. © 2023 John Wiley & Sons, Inc. Published 2023 by John Wiley & Sons, Inc.
SS)-DATAN
Scheme 1.1 Derivatization reaction of HAs and AAs either with (RR)-DATAN or with (SS)-DATAN for producing diastereomers. Source: Oliveira et al. [2]/with permission of MDPI/Public Domain CC BY 4.0.
Back in the 1980s when the chiral columns started to be commercialized, it was believed that the enantioresolution depended only on the CSP and, thus, the mobile phases and organic modifiers were chosen based only on solubility parameters of the chiral solutes.
Moreover, most of the chiral selectors had elution mode restrictions, and thus, it was usual to change the chiral selector without exploring the mobile phase. Nowadays, the interdependent role of chiral selectors and mobile phases is acknowledged.
The illustrated pattern in Figure 1.1 should be considered for starting a direct chiral separation.
Nevertheless, in method development one should have in mind the intended use. The reason is that the application imposes restrictions. For instance, for measuring enantiomeric ratios of synthetic products one can easily use the normal elution mode, but if the application is for determining enantiomeric fraction in environmental matrices, which implies mass spectrometer detection, the normal elution mode is impaired, and the chromatographic conditions should be developed under polar organic or reversed phase elution modes [1]. Thus, for selecting the separation conditions one should bare this in mind.
The influence of the elution mode and organic modifier can be perceived in the enantiomeric separation of the proton pump inhibitors: omeprazole, lansoprazole, and pantoprazole in four different polysaccharide-based chiral selectors [5]. Taking as an example, the separation values (enantioselectivity [α] and resolution [Rs]) obtained for omeprazole at the CSPs
Buffer/additives
CSPs
Elution mode
Organic modifier
Temperature
Application
Figure 1.1 Illustration of a pattern that should be considered for starting a direct chiral separation.
tris-(3,5-dimethylphenylcarbamate) and tris-[(S)-1-phenylethylcarbamate] of amylose were α = 1.57 and Rs = 3.15 and α = 1.48 and Rs = 1.71, respectively, using Hex:EtOH (70/30, v/v) as mobile phase, while at tris-(3,5dimethylphenylcarbamate) of cellulose no separation was obtained in any of the elution conditions examined. The graphic at Figure 1.2 [5] illustrates the influence of the elution mode in the separation of omeprazole in going from polar organic to reverse phase at the CSP of tris- (3,5dimethylphenylcarbamate) of amylose.
At the end of the 1980s, the high number of commercial chiral columns available demanded a classification of the chiral selectors. The classification was then based on their recognition interactions or, in other words, in the formation of solute–CSP complexes. The CSPs were
Figure 1.2 Graphic showing the retention factors (k1 and k2) of omeprazole enantiomers at tris-(3,5-dimethylphenylcarbamate) of amylose in going from neat MeCN to aqueous MeCN solutions as mobile phases. Source: Cass et al. [5]/with permission of Taylor & Francis.
classified in five categories (Types I–V) [6, 7]. Variations of these classifications have been made considering the chiral selectors in three main groups: macromolecular selectors, macrocyclic selectors, and lowmolecular mass selectors [8].
Although the three-point interaction model, as elaborated by Dalgliesh for paper chromatography separation of AAs [9], didactically explains the formation of the transient diastereomeric complex between solute and CSP, the interactions accountable for the chiral discrimination still demand clarification [8]. Moreover, a CSP encompasses several heterogeneous non- selective and stereoselective active sites that contribute to the resolution of an enantiomeric mixture by the CSP at a given mobile phase and temperature [10].
It is important to stress that, due to the complexity of chiral discrimination mechanisms, there is no preset- up condition for achieving resolution for a given application. For method development, understanding the most important interactions should somehow help in a planned workflow.
The most important types of CSPs and their properties will be discussed in detail throughout the chapters of this book.