Table Of Content❦
INTERPRETATION OF MS–MS MASS
SPECTRA OF DRUGS AND PESTICIDES
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WILEY SERIES ONMASS SPECTROMETRY
Series Editors
Dominic M. Desiderio
Departments of Neurology and Biochemistry University of Tennessee Health Science Center
Joseph A. Loo
Department of Chemistry and Biochemistry UCLA
Founding Editors
Nico M. M. Nibbering (1938–2014)
Dominic M. Desiderio
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INTERPRETATION OF MS–MS MASS
SPECTRA OF DRUGS AND PESTICIDES
WILFRIED M. A. NIESSEN
hyphen MassSpec, The Netherlands
RICARDO A. CORREA C.
Trans-Laboratory, Brussels, Belgium
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This edition frst published 2017
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Library of Congress Cataloging-in-Publication Data:
Names: Niessen, W. M. A. (Wilfried M. A.), 1956- author. | Correa C., Ricardo
A., 1961- author.
Title: Interpretation of MS-MS mass spectra of drugs and pesticides /
Wilfried M.A. Niessen, Ricardo A. Correa C.
Other titles: Wiley-Interscience series on mass spectrometry.
Description: Hoboken, New Jersey : John Wiley & Sons, 2016. | Series: Wiley
series on mass spectrometry | Includes bibliographical references and
index.
Identifers: LCCN 2016031593 (print) | LCCN 2016046526 (ebook) | ISBN
9781118500187 (cloth) | ISBN 9781119294245 (pdf) | ISBN 9781119294252
(epub)
Subjects: LCSH: Tandem mass spectrometry. | Liquid chromatography. |
Drugs–Analysis. | Pesticides–Analysis.
Classifcation: LCC QD96.M3 N525 2016 (print) | LCC QD96.M3 (ebook) | DDC
543/.65–dc23
LC record available at https://lccn.loc.gov/2016031593
Cover Design by Wiley.
Cover image: Curtsey of the authors
Set in 10/12pt, TimesLTStd by SPi Global, Chennai, India.
Printed in the United States of America
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CONTENTS
Preface xi
Abbreviations xiii
1 Introduction to LC–MS Technology 1
1.1 Introduction 2
1.2 Analyte Ionization: Ion Sources 3
❦ 1.2.1 Electron Ionization 3 ❦
1.2.2 Chemical Ionization 7
1.2.3 Atmospheric-Pressure Ionization 13
1.2.4 Electrospray Ionization 16
1.2.5 Atmospheric-Pressure Chemical Ionization and Photoionization 20
1.2.6 Other Ionization Techniques 26
1.3 Mass Spectrometer Building Blocks 28
1.3.1 Introduction 28
1.3.2 Quadrupole Mass Analyzer 32
1.3.3 Ion-Trap Mass Analyzer 33
1.3.4 Time-of-Flight Mass Analyzer 34
1.3.5 Orbitrap Mass Analyzer 35
1.3.6 Other Mass Analyzers 36
1.4 Tandem Mass Spectrometry 37
1.4.1 Introduction 37
1.4.2 Tandem Quadrupole Instruments 39
1.4.3 Ion-Trap Instruments 40
1.4.4 Quadrupole–Linear Ion-Trap Hybrid Instruments 41
1.4.5 Quadrupole–Time-of-Flight Hybrid Instruments 41
1.4.6 Orbitrap Hybrid Instruments for MS–MS and MSn 41
1.4.7 Other Instruments for MS–MS and MSn 42
1.4.8 MS–MS and MSn in the Analysis of Drugs and Pesticides 43
1.5 Data Acquisition 43
1.5.1 Introduction 43
1.5.2 Selected-Ion and Selected-Reaction Monitoring 44
1.5.3 Structure-Specifc Screening: Precursor-Ion and Neutral-Loss Analysis 44
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vi CONTENTS
1.5.4 Data-Dependent Acquisition 45
1.5.5 Data-Independent Acquisition 45
1.6 Selected Literature on Mass Spectrometry 45
References 46
2 Interpretation of Mass Spectra 55
2.1 Mass Spectrometry: A Nuclear Affair 55
2.2 Isomers, Isotones, Isobars, Isotopes 56
2.3 Masses in MS 58
2.4 Isotopes and Structure Elucidation 60
2.5 Nitrogen Rule, Ring Double-Bond Equivalent, and Hydrogen Rule 61
2.6 Resolving Power, Resolution, Accuracy 63
2.7 Calculating Elemental Composition from Accurate m/z 66
2.8 Protonated and Deprotonated Molecules and Adduct Ions 67
References 69
3 Fragmentation of Even-Electron Ions 71
3.1 Introduction 72
3.2 Analyte Ionization Revisited 73
3.3 Fragmentation of Odd-electron Ions 74
3.4 High-energy Collisions of Protonated Molecules 76
3.4.1 General Aspects 77
3.4.2 Selected Examples 79
3.5 Fragmentation of Protonated Molecules 81
3.5.1 Singly-Charged Peptides 82
3.5.2 Protonated Small Molecules: Complementary Fragment Ions 83
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3.5.3 Fragmentation of Peptides Revisited 86
3.5.4 Direct-Cleavage Reactions 89
3.5.5 Consecutive Small-Molecule Losses 89
3.5.6 Other Fragmentation Reactions 89
3.5.7 Loss of Radicals from Even-Electron Ions 91
3.5.8 Skeletal Rearrangements in Protonated Molecules 92
3.6 Characteristic Positive-ion Fragmentation of Functional Groups 94
3.6.1 Cleavages of C—C Bonds 94
3.6.2 Alcohols and Ethers 95
3.6.3 Aldehydes and Ketones 96
3.6.4 Carboxylic Acids and Esters 96
3.6.5 Amines and Quaternary Ammonium Compounds 98
3.6.6 Amides, Sulfonyl Ureas, and Carbamates 99
3.6.7 Compounds Containing Phosphorus or Sulfur 101
3.6.8 Miscellaneous Compound Classes 101
3.7 Fragmentation of Deprotonated Molecules 102
3.7.1 High-Energy CID with NICI-Generated Deprotonated Molecules 102
3.7.2 General Aspects 102
3.7.3 Alcohols and Ethers 102
3.7.4 Carboxylic Acid and Esters 103
3.7.5 Amines 104
3.7.6 Amides and Ureas 104
3.7.7 Sulfonamides and Related Sulfones 106
3.7.8 Halogenated Compounds 107
3.7.9 Miscellaneous Compound Classes 108
3.8 Fragmentation of Metal-ion Cationized Molecules 108
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CONTENTS vii
3.9 Generation of Odd-electron Ions in ESI-MS, APCI-MS, and APPI-MS 111
3.10 Useful Tables 112
References 122
4 Fragmentation of Drugs and Pesticides 129
4.1 Fragmentation of Drugs for Cardiovascular Diseases and Hypertension 134
4.1.1 β-Blockers or β-Adrenergic Antagonists 134
4.1.2 Dihydropyridine Calcium Antagonists 137
4.1.3 Angiotensin-Converting Enzyme Inhibitors 140
4.1.4 Diuretic Drugs 142
4.1.5 Angiotensin II Receptor Antagonists 148
4.1.6 Other Antihypertensive Compounds 149
4.1.7 Antiarrhythmic Agents 153
References 155
4.2 Fragmentation of Psychotropic or Psychoactive Drugs 158
4.2.1 Phenothiazines 158
4.2.2 Other Classes of Neuroleptic Drugs 161
4.2.3 Antidepressants 163
4.2.4 Benzodiazepines 167
4.2.5 Local Anesthetics 173
4.2.6 Barbiturates 173
4.2.7 Anticonvulsant Drugs 174
4.2.8 Other Psychotropic Drugs 175
References 176
4.3 Fragmentation of Analgesic, Antipyretic, and Anti-Infammatory Drugs 179
4.3.1 Acetaminophen 179
❦ 4.3.2 Salicylic Acid Derivatives 179 ❦
4.3.3 Nonsteroidal Anti-Infammatory Drugs 180
4.3.4 COX-2 Inhibitors 184
References 186
4.4 Fragmentation of Drugs Related to Digestion and the Gastrointestinal Tract 188
4.4.1 Antidiabetic Drugs 188
4.4.2 Antiulcer Drugs 191
4.4.3 Lipid-Lowering Agents 192
4.4.4 Anorexic Drugs 195
4.4.5 Antiemetic Drugs 196
References 197
4.5 Fragmentation of Other Classes of Drugs 199
4.5.1 𝛽-Adrenergic Receptor Agonists 199
4.5.2 Histamine Antagonists 199
4.5.3 Anticholinergic Agents 202
4.5.4 Drugs against Alzheimer’s Disease: Acetylcholinesterase Inhibitors 204
4.5.5 Antiparkinsonian Drugs 208
4.5.6 Antineoplastic and Cytostatic Drugs 209
4.5.7 Immunosuppressive Drugs 214
4.5.8 X-ray Contrast Agents 216
4.5.9 Anticoagulants and Rodenticides 218
4.5.10 Conclusions 219
References 220
4.6 Fragmentation of Steroids 222
4.6.1 Introduction 222
4.6.2 Fragmentation of Steroids 223
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4.6.3 Fragmentation in 3-Keto-Δ -Steroids 224
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4.6.4 Anabolic Steroids 225
4.6.5 Progestogens 227
4.6.6 Corticosteroids 228
4.6.7 Estrogens 234
4.6.8 Steroid Conjugates 236
References 237
4.7 Fragmentation of Drugs of Abuse 241
4.7.1 Introduction 241
4.7.2 Amphetamine and Related Compounds 241
4.7.3 Cannabinoids 248
4.7.4 Cocaine and Related Substances 250
4.7.5 Opiates 251
4.7.6 Miscellaneous Drugs of Abuse 252
4.7.7 Designer Drugs 255
References 258
4.8 Fragmentation of Antimicrobial Compounds 262
4.8.1 Sulfonamides 262
4.8.2 Chloramphenicol and Related Compounds 265
4.8.3 𝛽-Lactams 266
4.8.4 (Fluoro)quinolones 273
4.8.5 Aminoglycosides 277
4.8.6 Tetracyclines 279
4.8.7 Nitrofurans 279
4.8.8 Macrolides 281
4.8.9 Miscellaneous Antibiotics 282
References 284
4.9 Fragmentation of Antimycotic and Antifungal Compounds 289
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4.9.1 Imidazolyl Antimycotic Compounds 289
4.9.2 Triazolyl Antifungal Compounds 290
4.9.3 Benzamidazole Fungicides 292
4.9.4 Other Classes of Fungicides 293
References 296
4.10 Fragmentation of Other Antibiotic Compounds 297
4.10.1 Anthelmintic Drugs 297
4.10.2 Antiprotozoal, Coccidiostatic, and Antimalarial Agents 299
4.10.3 Antiviral Drugs 304
4.10.4 Antiseptics and Disinfectants 309
References 312
4.11 Pesticides 315
4.11.1 Triazine Herbicides 315
4.11.2 Carbamates 317
4.11.3 Quaternary Ammonium Herbicides 322
4.11.4 Organophosphorus Pesticides 322
4.11.5 Urea Herbicides: Phenylureas, Benzoylphenylureas, and Others 329
4.11.6 Sulfonylurea Herbicides 334
4.11.7 Chlorinated Phenoxy Acid Herbicides 337
4.11.8 Phenolic Compounds 339
4.11.9 Miscellaneous Herbicides 339
References 345
5 Identifcation Strategies 351
5.1 Introduction 351
5.2 Confrmation of Identity in Following Organic Synthesis 352
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CONTENTS ix
5.3 Confrmation of Identity in Targeted Screening by SRM-based Strategies 353
5.3.1 Environmental Analysis 354
5.3.2 Food Safety Analysis 355
5.3.3 Sports Doping Analysis 355
5.3.4 General Unknown Screening in Toxicology 355
5.4 Confrmation of Identity by High-resolution Accurate-mass MS Strategies 356
5.4.1 Environmental and Food Safety Analysis 356
5.4.2 General Unknown Screening in Toxicology 357
5.4.3 Sports Doping Analysis 357
5.5 Library Searching Strategies in Systematic Toxicological Analysis 357
5.6 Dereplication and Identifcation of Natural Products and Endogenous Compounds 360
5.7 Identifcation of Structure-related Substances 361
5.7.1 Drug Metabolites 362
5.7.2 Impurities and Degradation Products 366
5.8 Identifcation of Known Unknowns and Real Unknowns 366
References 370
Compound Index 381
Subject Index 389
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PREFACE
In the 1980s, tandem mass spectrometry was introduced for University Hospital of Limoges, France) for providing his
the structural elucidation of even-electron ions (protonated mass spectral library of negative-ion mass spectra; and Dr
or deprotonated molecules) generated by soft ionization Bernhard Wüst of Agilent Technologies for his help with us-
techniques such as fast-atom bombardment, thermospray, ing the Agilent Broecker, Herre & Pragst PCDL for forensic
and electrospray. When compared to the fragmentation of toxicology. The information from these libraries and other
odd-electron ions generated by electron ionization, scientists data sets is complemented by data from the scientifc litera-
were well aware of the fact that different rules apply to ture.
❦ the fragmentation of even-electron ions. Surprisingly, no The origins of this book can be found in two publica- ❦
major fundamental research was carried out on trying to tions describing the fragmentation of toxicologically rele-
understand and describe these differences. More effort was vant drugs in both positive-ion tandem mass spectrometry
placed on the development of improved instrumentation (Niessen, 2011) and negative-ion tandem mass spectrometry
and advanced applications for the emerging technologies. (Niessen, 2012). Soon after, the authors decided to develop
This particular effort paid off, as exemplifed by tandem the project further by extending the number of compounds
mass spectrometry which, often in combination with gas covered and the detail of the information provided. The frag-
or liquid chromatography, has been a major contributor to mentation of some 1300 compounds and the product-ion
the progress of many scientifc disciplines, for example, mass spectra of even more are studied and interpreted in
pharmaceutical, biochemical, and environmental sciences; this book.
food safety; sports doping analysis; clinical diagnostics; This volume consists of fve chapters. Chapters 3 and 4
forensics; and toxicology. are the main chapters, where proposed fragmentation rules
This work is an attempt to add to the understanding of the for the “Fragmentation of Even-Electron Ions” (Chapter 3)
fragmentation of even-electron ions. This has been done by are derived from the behavior of the “Fragmentation of
studying the fragmentation of a wide variety of compounds, Drugs and Pesticides” (Chapter 4) pertaining to many dif-
with a special focus on chemical structure similarities, that ferent classes of compounds. Chapter 1, “Introduction to
is, from the same class. The basic data set used comprises LC–MS–MS Technology”, provides a concise introduction
a number of mass spectral libraries developed for general to mass spectrometry technology. Chapter 2, “Interpretation
unknown screening in toxicology. In this respect, we need of Mass Spectra” gives the basic concepts and defnitions
to thank Dr Wolfgang Weinmann (originally at the Insti- related to the information that can be extracted from mass
tute of Legal Medicine, University of Freiburg, Germany, spectra. Finally, Chapter 5, “Identifcation Strategies” gives
and currently at the Institute of Forensic Medicine, Uni- an overview of the different classes of unknowns and iden-
versity of Bern, Switzerland) for providing public access tifcation strategies that exist as well as how they relate to
to his toxicology library and the library of designer drugs multiple areas of application.
via the Internet (http://www.chemicalsoft.de/index.html); Dr Last but not least, special thanks go to our families, and
Pierre Marquet (of the Faculty of Medicine, Department of themany peoplewho have inspired us to continueworking on
Pharmacology, Toxicology, and Pharmacovigilance at the this project.We hope that you, as our reader, fnd thismaterial
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