Table of Contents

Cover

Title Page

Wiley Series on Mass Spectrometry

Dedication

Preface

List of Contributors

About the Editors

Chapter 1: Basic Considerations for the Analyst for Veterinary Drug Residue Analysis in Animal Tissues

1.1 Introduction
1.2 Pharmacokinetics
1.3 Metabolism and Distribution
1.4 Choice of Analytical Method
1.5 Importance of Regulatory Limits
1.6 International Obligations for Regulatory Analytical Laboratories
1.7 Conclusions
References

Chapter 2: Emerging Techniques in Sample Extraction and Rapid Analysis

2.1 Introduction
2.2 Sample Extraction
2.3 Extract Clean-up with Solid-Phase Sorbents
2.4 Micro-extraction Techniques for Solvent and Sorbent Extraction
2.5 Emerging Techniques in Liquid Chromatography
2.6 Direct Mass Spectrometry Analysis of Sample Extracts
2.7 Ion Mobility Spectrometry
2.8 Conclusions
References

Chapter 3: Capabilities and Limitations of High-Resolution Mass Spectrometry (HRMS): Time-of-flight and Orbitrap™

3.1 Available Technology
3.2 Capabilities and Limitations of the Technology as Compared to LC-MS/MS (Tandem Quadrupole Mass Spectrometer)
3.3 Analytical Methods for Veterinary Drug Residues
3.4 Doping Control
3.5 Accurate Mass MS in Research and Metabolism Studies
3.6 Designer Drugs and Generic Detection Strategies
3.7 The Future of Accurate Mass Spectrometry in Residue Analysis
References

Chapter 4: Hormones and β-Agonists

4.1 Introduction
4.2 Advances in Classical Analysis of Exogenous Synthetic Hormones
4.3 Bio-Based Screening Methods for Steroid Hormones, β-Agonists, and Growth Hormones
4.4 Natural Hormones
4.5 Control for Synthetic β-Agonists: Screening and Confirmatory Methods
References

Chapter 5: Analysis of Anthelmintic and Anticoccidial Drug Residues in Animal-Derived Foods

5.1 Introduction
5.2 Chemistry and Mode of Action
5.3 Legislation
5.4 Sample Preparation Protocols for Anti-parasitic Agents in Food Matrices
5.5 LC-MS and GC-MS Detection of Anti-parasitic Agents in Food
5.6 Conclusions
References

Chapter 6: Sedatives and Tranquilizers

6.1 Introduction
6.2 Classification and Representative Compounds
6.3 Use of Sedatives and Tranquilizers to Prevent Stress Syndrome during the Transport of Pigs to Slaughter
6.4 Sedatives and Tranquilizers with an Approved Veterinary Use in Food-Producing Animals
6.5 Sedatives and Tranquilizers without an Approved Veterinary Use in Food-Producing Animals
6.6 Cocktails
6.7 Issues of Environmental Contamination
6.8 Maximum Residue Limits (MRLs)
6.9 Systematic Veterinary Control over Residues and Surveillance Studies
6.10 Analyte Stability
6.11 Analytical Methods for Determination of Residues
6.12 Performance and Validation of the Analytical Methods
References

Chapter 7: The Use of Pyrethroids, Carbamates, Organophosphates, and Other Pesticides in Veterinary Medicine

7.1 Introduction
7.2 Veterinary Drug Properties, Structures, and Regulation
7.3 Toxicology, Pharmacokinetics, and Metabolism
7.4 Analytical Methods
7.5 Conclusion
References

Chapter 8: Non-steroidal Anti-inflammatory Drugs

8.1 Introduction: What Are Pain Killers (Analgesics) and NSAIDs?
8.2 Veterinary Drug Properties, Structures, and Regulation
8.3 Pharmacokinetics/Metabolism
8.4 Acceptable Daily Intake (ADI)
8.5 Maximum Residue Limits/Tolerances
8.6 Analysis of NSAID Residues in Food
8.7 Literature Reviews of Analytical Methods for NSAIDs in Biological Samples
8.8 New Developments in NSAIDs
8.9 Conclusion
References

Chapter 9: Certain Dyes as Pharmacologically Active Substances in Fish Farming and Other Aquaculture Products

9.1 Introduction
9.2 Therapeutic Applications and Chemistry of Certain Dyes Used in Fish Farming
9.3 Toxicological Issues
9.4 Regulatory Issues
9.5 Analytical Methods for Residue Control
9.6 Recent Trading Issues with Dye Alerts
9.7 Conclusions
References

Chapter 10: Method Validation and Quality Assurance/Quality Control Approaches for Multi-residue Methodsc

10.1 Introduction
10.2 Sources of Guidance on Method Validation
10.3 Practical Considerations
10.4 Examples of Validation Protocols for MRMs
10.5 Quality Assurance/Quality Control
10.6 Conclusion
References

Index

Wiley Series on Mass Spectrometry

End User License Agreement

List of Illustrations

Chapter 2: Emerging Techniques in Sample Extraction and Rapid Analysis

Figure 2.1 Filter vial dispersive solid-phase extraction.
Figure 2.2 Representation of restricted access material particle.
Figure 2.3 Direct analysis in real time (DART) and desorption electrospray ionization (DESI) sources for ambient mass spectrometry.
Figure 2.4 Laser diode thermal desorption source for rapid mass spectrometry.

Chapter 3: Capabilities and Limitations of High-Resolution Mass Spectrometry (HRMS): Time-of-flight and Orbitrap™

Figure 3.1 Schematic drawing of an orthogonal time-of-flight instrument.
Figure 3.2 Spread of kinetic energy of transmitted ions.
Figure 3.3 Schematic drawing of a modern Orbitrap™ instrument.
Figure 3.4 Effect on mass resolving power on the separation of exogenous from endogenous (matrix related) compounds.
Figure 3.5 Effect of reducing the mass extraction window on the selectivity of detection (marbofloxacin in liver extract).
Figure 3.6 Distortion of a chromatographic analyte signal by applying a too narrow mass extraction window (norfloxacin with 10,000 FWHM and 2 mDa mass extraction window).
Figure 3.7 Determination of detection related parameters such as LOD or CCα in a detection-noise-fee environment.
Figure 3.8 Steroid ionization under atmospheric pressure conditions, analytes lacking a 3-keto-4-ene in the A ring may require derivatization.
Figure 3.9 Accurate mass data and designer drugs, potential for direct analysis, or detection using either targeted or untargeted metabolomics.

Chapter 4: Hormones and β-Agonists

Figure 4.1 Possible sources of exogenous natural compounds can be from feed, injection (steroids or somatotropin), pour-on application, or conversion or degradation of compounds in the gastrointestinal tract.
Figure 4.2 Metabolism of RALs, dashed arrows represent conversion of the toxins toward zeranol.
Figure 4.3 Steroidogenesis, the steroid pathway showing relations between individual steroids.
Figure 4.4 Structures of NT esters. NT-acetate: R = CH₃, NT-benzoate: R = phenyl, NT-cypionate: C₂H₄-cyclopentyl, NT-hemisuccinate: CH₂-succinyl, NT-decanoate: R = C₉H₁₉, NT-undecanoate: R = C₁₀H₂₁, NT-laureaat: C₁₁H₂₃, NT-phenylpropionate: R = C₂H₄-phenyl, NT-propionate: R = C₂H₅.
Figure 4.5 Chemical structures of the NT metabolites: estranediol isomers.
Figure 4.6 Structures of 17β-boldenone (17β-Bol) and 17β-testosterone (17β-T) (above) and of androsta-1,4-diene-3,17-dione (ADD) and androst-4-ene-3,17-dione (AED).
Figure 4.7 The mechanism of converting the signal from the β-adrenergic receptor (βAR).

Chapter 5: Analysis of Anthelmintic and Anticoccidial Drug Residues in Animal-Derived Foods

Figure 5.1 Chemical structures of benzimidazoles, benzimidazole metabolites, and levamisole.
Figure 5.2 Chemical structures of tetrahydropyrimidines.
Figure 5.3 Chemical structures of some organophosphate anthelmintic agents and metabolites.
Figure 5.4 Chemical structures of some flukicide drugs.
Figure 5.5 Chemical structures of different macrocyclic lactone molecules.
Figure 5.6 Structure of miscellaneous anti-parasitic drugs and metabolites.
Figure 5.7 Chemical structures of ionophores.
Figure 5.8 Structures of chemical anticoccidials (and some of their metabolites) that are licensed as veterinary drugs,⁷⁴ feed additives⁷⁵, or pesticides⁷⁶ in the EU.
Figure 5.9 Structures of chemical anticoccidials that are not licensed in the EU.
Figure 5.10 Overview of modified QuEChERS sample preparation procedure for the determination of anthelmintic drug residues in milk samples.
Figure 5.11 Steps of the filter-vial dispersive-SPE process: (1) 25 mg C18 contained in shell portion of the filter vial, (2) addition of 0.4 ml of the extracts, (3) partial capping of the vials, (4) shaking of the tray for 30 s, (5) pressing of the vials, and (6) final extracts ready for analysis.
Figure 5.12 Overlay of analytes (a–c) at concentration of 2 µg/kg (OXY, CLOR, BITH, and MOR were 4 µg/kg) and selected internal standards (d–e).
Figure 5.13 Overlay of LC-MS/MS chromatograms for all 20 coccidiostat analytes (a–c) at concentrations of 2.5 µg/kg (HALO at 10 µg/kg; DNC and IMID at 25 µg/kg; TOL, TOL-SO, and TOL-SO₂ at 50 µg/kg) and internal standards (d), in a fortified avian muscle sample. (Abbreviations: IMID, imidocarb; ARP, arprinocid; ETHOP, ethopabate; TOL-SO, toltrazuril sulfoxide; DICLAZ, diclazuril; TOL, toltrazuril; DECOQ, decoquinate; SEMDUR, semduramicin; MAD, maduramicin; DIAV, diaveridine; HALO, halofuginone; ROB, robenidine; DNC, 4,4′-dinitrocarbanilide; MON, monensin; SAL, salinomycin; NAR, narasin; CLOP, clopidol; TOL-SO₂, toltrazuril sulfone; LAS, lasalocid; LAID, laidlomycin).

Chapter 6: Sedatives and Tranquilizers

Figure 6.1 Chemical structures of selected sedatives and tranquilizers.

Chapter 7: The Use of Pyrethroids, Carbamates, Organophosphates, and Other Pesticides in Veterinary Medicine

Figure 7.1 Organochlorine pesticide structures.
Figure 7.2 Typical structures of pyrethrins and pyrethroids showing chiral centers.
Figure 7.3 Structures of some typical organophosphates.
Figure 7.4 Structures of some typical carbamates.

Chapter 9: Certain Dyes as Pharmacologically Active Substances in Fish Farming and Other Aquaculture Products

Figure 9.1 World capture fisheries and aquaculture production per year from FAO (2014).¹
Figure 9.2 International seafood trade flows from the main importers and exporters by year and origin of production (aquaculture vs fisheries). The size of the circles is proportional to the value of the exports. The shading is based on the percentage represented by aquaculture of the total fish production in the exporting country in contrast to wild catch fisheries (10 equal intervals between 0% and 100%). The lighter the grey shading, the more aquaculture; the darker the grey shading, the more product is from wild catch fisheries.
Figure 9.3 Structures of pharmacologically active dyes.
Figure 9.4 EU alerts for dye residues in fin-fish aquaculture. Extracted from the EU RASFF website over the period 2002–2016.²¹³
Figure 9.5 EU alerts for dye residues in fin-fish aquaculture products sorted per countries of origin. Extracted from the EU RASFF website over the 2002–2016 period.²¹³
Figure 9.6 EU alerts for MG residues in fin-fish farming products sorted per year. Extracted from EU RASFF website over the period 2004–2015.²¹³

List of Tables

Chapter 4: Hormones and β-Agonists

Table 4.1 Summary of representative confirmatory methods based on LC and GC for the control of hormonal active compounds in food-producing animals in 2012–2014
Table 4.2 The main characteristics of reported LC-MS assays to identify and/or quantify GHRPs in urine and plasma
Table 4.3 Most relevant β-agonists for residue control
Table 4.4 Adrenergic receptor types, subtypes, and their effects in organism

Chapter 5: Analysis of Anthelmintic and Anticoccidial Drug Residues in Animal-Derived Foods

Table 5.1 Tolerances and limits for anticoccidial residues in edible tissues
Table 5.2 Maximum residue limits for anthelmintic veterinary drugs listed under Council Regulation 37/2010/EC.⁷⁴

Chapter 6: Sedatives and Tranquilizers

Table 6.1 Maximum residue limits (MRLs) of sedatives and tranquilizers (µg/kg), established by the Codex Alimentarius, the European Union, and some of the world's largest food-producing/importing countries
Table 6.2 Current (published since 2000) and selected earlier analytical methods for determination of sedatives and tranquilizers in food-producing animals and derived products

Chapter 7: The Use of Pyrethroids, Carbamates, Organophosphates, and Other Pesticides in Veterinary Medicine

Table 7.1 Approximate dates for the pyrethroid generations
Table 7.2 Extraction and detection methods for pesticides approved for use in veterinary medicine

Chapter 8: Non-steroidal Anti-inflammatory Drugs

Table 8.1 Classification of the NSAIDs (number in Table corresponds to subsection in text where compound is discussed)
Table 8.2 Maximum residue limits (MRLs) established for NSAIDs by the European Union
Table 8.3 Maximum residue limits (MRLs) established for NSAIDs by the Australian Pesticides and Veterinary Medicines Authority (APVMA).⁹³
Table 8.4 Maximum residue limits (MRLs) established for NSAIDs by Health Canada.⁹⁴
Table 8.5 Non-multiresidue methods for NSAIDs in animal-derived food matrices^a
Table 8.6 Multi-residue methods for NSAIDs in biological matrices

Chapter 9: Certain Dyes as Pharmacologically Active Substances in Fish Farming and Other Aquaculture Products

Table 9.1 Overview of specific regulations for dyes in aquaculture products by countries
Table 9.2 Percentage of veterinary drugs (dyes) violations collected over 2000–2009 period by seafood type and by inspection body
Table 9.3 Percentage of veterinary drugs (dyes) violations by seafood type aquaculture. Extracted from the EU RASFF website over the period 2002–2016.²¹³

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Published simultaneously in Canada

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Library of Congress Cataloging-in-Publication Data

Names: Kay, Jack F., editor. | MacNeil, James D., editor. | Wang, Jian, 1964-editor.

Title: Chemical analysis of non-antimicrobial veterinary drug residues in food / edited by Jack F. Kay, James D. MacNeil, Jian Wang.

Description: Hoboken, New Jersey : John Wiley & Sons, Inc., [2017] | Includes bibliographical references and index.

Identifiers: LCCN 2016033527| ISBN 9781118695074 (cloth) | ISBN 9781119325901 (epub)

Subjects: LCSH: Veterinary drug residues. | Food contamination.

Classification: LCC RA1270.V47 C4425 2017 | DDC 664/.07–dc23 LC record available at https://lccn.loc.gov/2016033527

WILEY SERIES ON MASS SPECTROMETRY

Departments of Neurology and Biochemistry
University of Tennessee Health Science Center

Preface

Food safety continues to be a topic of great interest to consumers and is frequently a topic for media discussion. However, what is not routinely reported is the vast effort by many national, regional, and international bodies and scientists – both governmental and independent – to ensure that food production and trade do not place consumers at risk while ensuring a continuous supply of wholesome food.

A key aim of regulating the use of veterinary drugs is to ensure that authorized products are used responsibly in animals and that their residues in food of animal origin do not pose unacceptable health risks to consumers. To assist in this process, robust and validated analytical methods to detect a wide range of potential residues in food matrices are required.

The earlier volume in this series, Chemical Analysis of Antibiotic Residues in Food, was published in 2012 and set out in detail how drug safety is considered and limits are set for their residues in foods. It also described how residue monitoring programs are established and checked to ensure sound results are generated to inform necessary regulatory actions to protect consumers. These topics are generic and apply equally to antibiotics and other veterinary drug classes. The companion volume to this current book also provided detailed information on analytical methods for antibiotic residues.

The purpose of this current book, Chemical Analysis of Non-antimicrobial Veterinary Drug Residues, is to update readers on developments in technology and approaches since the publication of the earlier volume. It also seeks to expand the coverage of veterinary drug residues to all other key areas of veterinary drug treatments, thus providing a comprehensive two-volume set for reference and training purposes.

The main themes of the book include detailed discussions on emerging technologies (Chapter 2); high resolution mass spectrometry and related techniques (Chapter 3); hormones and β-agonists (Chapter 4); anthelmintics (Chapter 5); sedatives and tranquilizers (Chapter 6); pyrethroids, carbamates, organophosphates, and other pesticides used in veterinary medicines (Chapter 7); non-steroidal anti-inflammatories (Chapter 8); dyes (Chapter 9); and developments in the validation of multi-class multi-residue methods and related quality control/quality assurance issues (Chapter 10).

The editors and authors of this book are internationally recognized experts and leading scientists with extensive personal experience in preparing food safety regulations and/or in the chemical analyses of veterinary drug residues in food of animal origin. This book offers a valuable and up to date view of the science in this area. It has been deliberately written and organized to complement and update where necessary the information contained in the earlier companion volume. The editors hope that this volume completes and addresses the need for readers from regulatory backgrounds and analytical laboratory staff to have a cutting-edge reference and training resource for the residues of all veterinary drug residues in food of animal origin.

26 August, 2016

Jack F. Kay
University of Strathclyde,
Glasgow, Scotland

James D. MacNeil
St. Mary's University,
Halifax, Canada

Jian Wang
Canadian Food Inspection Agency,
Calgary, Canada

List of Contributors

Christine Akre

Saskatoon Laboratory

Canadian Food Inspection Agency

Saskatoon, Saskatchewan

Canada

Wendy C. Andersen

Animal Drugs Research Center

US Food and Drug Administration

Denver, Colorado

USA

Marco H. Blokland

RIKILT

Part of Wageningen UR

Wageningen

The Netherlands

Joe O. Boison

Saskatoon Laboratory

Canadian Food Inspection Agency

Saskatoon, Saskatchewan

Canada

Toine Bovee

RIKILT

Part of Wageningen UR

Wageningen

The Netherlands

Andrew Cannavan

Food and Environmental Protection Laboratory

Joint FAO/IAEA Division on Nuclear Techniques in Food and Agriculture

International Atomic Energy Agency

Vienna

Austria

Vesna Cerkvenik-Flajs

Veterinary Faculty

Department of Pathology and Administrative Veterinary Medicine

Laboratory for Ecotoxicology and Forensic Veterinary Medicine

University of Ljubljana

Ljubljana

Slovenia

Alan Chicoine

Department of Veterinary Biomedical Sciences

Western College of Veterinary Medicine

University of Saskatchewan

Saskatoon, Saskatchewan

Canada

Martin Danaher

Food Safety Department

Teagasc Food Research Centre

Ashtown

Dublin 15

Ireland

Ambrose Furey

Department of Chemistry

Cork Institute of Technology (CIT)

Bishopstown

Cork

Ireland

Zora Jandrić

Food and Environmental Protection Laboratory

Joint FAO/IAEA Division on Nuclear Techniques in Food and Agriculture

International Atomic Energy Agency

Vienna

Austria

Anton Kaufmann

Official Food Control Authority of the Canton of Zürich

Fehrenstrasse

Zürich

Switzerland

Jack F. Kay

Department of Mathematics & Statistics

University of Strathclyde

Glasgow, Scotland

United Kingdom

Jack J. Lohne

Animal Drugs Research Center

US Food and Drug Administration

Denver, Colorado

USA

James D. MacNeil

Department of Chemistry

St Mary's University

Halifax, Nova Scotia

Canada

Jelka Pleadin

Laboratory for Analytical Chemistry

Croatian Veterinary Institute

Zagreb

Croatia

Fernando Ramos

Pharmacy Faculty

Coimbra University

Coimbra

Portugal

Nathalie G.E. Smits

RIKILT

Part of Wageningen UR

Wageningen

The Netherlands

Saskia S. Sterk

RIKILT

Part of Wageningen UR

Wageningen

The Netherlands

Philip Teale

LGC Ltd

Fordham

Cambridgeshire

Sarah Tuck

Food Safety Department

Teagasc Food Research Centre

Dublin 15

Ireland

Sherri B. Turnipseed

Animal Drugs Research Center

US Food and Drug Administration

Denver, Colorado

USA

Leendert A. van Ginkel

RIKILT

Part of Wageningen UR

Wageningen

The Netherlands

Eric Verdon

Laboratory of Fougères

EU Reference Laboratory for Antimicrobial and Dye Residues in Food French Agency for Food Environmental and Occupational Health Safety

Fougères

France

Ana Vulić

Laboratory for Analytical Chemistry

Croatian Veterinary Institute

Zagreb

Croatia

About the Editors

Dr. Jack F. Kay received his Ph.D. from the University of Strathclyde, Glasgow, Scotland in 1980 and has been involved with veterinary drug residue analyses since 1991. He worked for the UK Veterinary Medicines Directorate to provide scientific advice on residue monitoring programs and managed the R&D program until his early retirement in September 2014. He helped draft Commission Decision 2002/657/EC and is an ISO trained assessor for audits to ISO 17025. He was co-chair of the CCRVDF ad hoc Working Group on Methods of Sampling and Analysis and steered Codex Guideline CAC/GL 71-2009 to completion after Dr. MacNeil retired. He co-chaired work extending this to cover multi-residue method performance criteria. He assisted JECFA in preparing an initial consideration of setting MRLs in honey and then took this forward for the CCRVDF. He also holds an Honorary Senior Research Fellowship in the Department of Mathematics and Statistics at the University of Strathclyde.

Dr. James D. MacNeil received his Ph.D. from Dalhousie University, Halifax, NS, Canada in 1972 and worked as a government scientist until his retirement in 2007. From 1982 to 2007 he was Head, Centre for Veterinary Drug Residues, now part of the Canadian Food Inspection Agency. He has served as a member of the Joint FAO/WHO Expert Committee on Food Additives (JECFA), co-chair of the working group on methods of Analysis and Sampling, Codex Committee on Residues of Veterinary Drugs in Foods (CCRVDF), is the former scientific editor for “Drugs, Cosmetics & Forensics” of J. AOAC Int., worked on IUPAC projects, has participated in various consultations on method validation, and is the author of numerous publications on veterinary drug residue analysis. He is a former General Referee for methods for veterinary drug residues for AOAC International and was appointed scientist emeritus by CFIA in 2008. He holds an appointment as an adjunct professor in the Department of Chemistry, St. Mary's University, Halifax, Canada, and has served as a part-time consultant to the JECFA Secretariat of the Food and Agriculture Organization of the United Nations since 2012.

Dr. Jian Wang received his Ph.D. at the University of Alberta in Canada in 2000, and then worked as a Postdoctoral Fellow at the Agriculture and Agri-Food Canada in 2001. He has been working as a leading research scientist at the Calgary Laboratory with the Canadian Food Inspection Agency since 2002. His scientific focus is on method development and validation using LC-MS/MS, UHPLC-QTOF, and UHPLC-Q Orbitrap for analyses of chemical residues including antibiotics, pesticides, and other chemical contaminants in foods. He also develops statistical approaches to estimating the measurement uncertainty based on method validation and quality control data using SAS program.