Genetic Toxicology Testing

A Laboratory Manual

1st Edition - May 26, 2016
Editor: Ray Proudlock
Language: English
Paperback ISBN:
9 7 8 - 0 - 1 2 - 8 0 0 7 6 4 - 8
eBook ISBN:
9 7 8 - 0 - 1 2 - 8 0 1 0 0 6 - 8

Genetic Toxicology Testing: A Laboratory Manual presents a practical guide to genetic toxicology testing of chemicals in a GLP environment. The most commonly used assays are descr… Read more

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Genetic Toxicology Testing: A Laboratory Manual
presents a practical guide to genetic toxicology testing of chemicals in a GLP environment. The most commonly used assays are described, from laboratory and test design to results analysis. In a methodical manner, individual test methods are described step-by-step, along with equipment, suggested suppliers, recipes for reagents, and evaluation criteria.

An invaluable resource in the lab, this book will help to troubleshoot any assay problems you may encounter to optimise quality and efficiency in your genetic toxicology tests. Genetic Toxicology Testing: A Laboratory Manual is an essential reference for those new to the genetic toxicology laboratory, or anyone involved in setting up their own.

List of Contributors
Foreword
Preface
- References
Chapter 1. A Practical Guide to Genetic Toxicology Testing
- Abstract
- 1.1 Introduction
- References
Chapter 2. General Recommendations
- Abstract
- 2.1 Establishing a New Assay
- 2.2 Spreadsheets and Manipulation of Results
- 2.3 Laboratory Historical Control Databases
- 2.4 Use of Computer Systems
- 2.5 Study Design
- 2.6 Evaluation Criteria
- 2.7 Organization of SOPs
- 2.8 Planning a Study
- 2.9 Preparing a Protocol Complying with GLP
- 2.10 Collecting Results
- 2.11 Reports
- 2.12 Training
- 2.13 Improving Quality and Efficiency
- 2.14 QA
- 2.15 Qualifying a Contract Laboratory
- 2.16 Responsibilities of the Study Monitor
- References
Chapter 3. Formulation of Test Articles
- Abstract
- 3.1 Introduction
- 3.2 Formulation Laboratories
- 3.3 Safety Data Sheets
- 3.4 Receipt of the Test Article
- 3.5 Formulation Types and Planning
- 3.6 Solubility and In Vitro Compatibility Testing
- 3.7 Formulation of Dose Solutions
- 3.8 Formulation of Bulk Formulations
- 3.9 Formulation of Suspensions
- 3.10 Chemical Analysis and Stability
- References
Chapter 4. The Bacterial Reverse Mutation Test
- Abstract
- 4.1 Introduction
- 4.2 History
- 4.3 Fundamentals
- 4.4 Equipment
- 4.5 Consumables
- 4.6 Reagents and Recipes
- 4.7 Suggested Phases in Development of the Test
- 4.8 The Bacterial Strains
- 4.9 Routine Testing
- 4.10 Standard Test Procedures
- 4.11 Screening Tests
- 4.12 Appendix 1: Growing and Monitoring Suspension Cultures
- References
Chapter 5. The Mouse Lymphoma TK Assay
- Abstract
- 5.1 Introduction
- 5.2 History
- 5.3 Provenance of the Cells
- 5.4 Spontaneous Mutation Frequency
- 5.5 Materials
- 5.6 Study Design
- 5.7 Evaluation Criteria
- 5.8 Predictivity of the MLA
- References
Chapter 6. The In Vitro Micronucleus Assay
- Abstract
- 6.1 Introduction
- 6.2 Practical Considerations
- 6.3 Methods
- 6.4 Materials
- 6.5 Protocols
- 6.6 Flow Cytometric Method
- References
Chapter 7. The In Vitro Chromosome Aberration Test
- Abstract
- 7.1 Introduction
- 7.2 History
- 7.3 Fundamentals
- 7.4 Equipment
- 7.5 Consumables and Reagents
- 7.6 Reagents and Recipes
- 7.7 Phases in Development of the Test
- 7.8 Cell Characterization
- 7.9 Routine Testing
- 7.10 Standard Test Procedures
- 7.11 Interpretation of Results
- 7.12 Screening Versions of the Test
- 7.13 Automation
- References
Chapter 8. The In Vivo Rodent Micronucleus Assay
- Abstract
- 8.1 Introduction
- 8.2 History
- 8.3 Fundamentals
- 8.4 Test Substance Considerations
- 8.5 Study Design
- 8.6 Manual Methods
- 8.7 Automated Analysis and Flow Cytometry
- 8.8 Study Validity
- 8.9 Interpretation of Results and Statistical Analysis
- 8.10 Nongenotoxic Mechanisms of Induction of Micronucleated Erythrocytes
- 8.11 Limitations of the Rodent Erythrocyte Micronucleus Test
- References
Chapter 9. The Rodent Bone Marrow Chromosomal Aberration Test
- Abstract
- 9.1 Introduction
- 9.2 History
- 9.3 Related Methods
- 9.4 Study Design and Performance
- 9.5 Terminal Procedures
- Appendix
- References
Chapter 10. The In Vivo Comet Assay Test
- Abstract
- 10.1 Introduction
- 10.2 The In Vivo Comet Assay in Regulatory Safety Testing
- 10.3 Fundamentals
- 10.4 Equipment and Nondisposable Supplies
- 10.5 Consumables
- 10.6 Reagents and Solutions
- 10.7 Test System
- 10.8 Study Design Considerations
- 10.9 Standard Test Procedures
- 10.10 Data and Reporting
- 10.11 Evaluating Unclear Results
- Acknowledgments
- References
Chapter 11. The Pig-a Endogenous Gene Mutation Assay
- Abstract
- 11.1 Introduction
- 11.2 History
- 11.3 Fundamentals
- 11.4 Study Design
- 11.5 Equipment
- 11.6 Consumables
- 11.7 Reagents and Recipes
- 11.8 Method Overview
- 11.9 Blood Collection
- 11.10 Leukodepletion and Platelet Removal
- 11.11 Sample Labeling
- 11.12 Column Separation and Sample Staining
- 11.13 Flow Cytometric Analysis: 96-Well Plate-Based Protocol
- 11.14 Tabulating and Summarizing Results
- 11.15 Evaluation and Interpretation of Results
- 11.16 Flow Cytometric Template Preparation
- 11.17 Example Plots
- 11.18 Storage and Shipment of Blood Samples
- 11.19 Aspiration of Postcolumn Samples
- References
Index

Ray Proudlock

Ray Proudlock graduated from Thames Polytechnic in Woolwich in 1974. His first laboratory jobs involved plant tissue culture and DNA turnover in cultured lymphocytes. Subsequently, he performed and supervised a wide range of genetic toxicology assays at Huntingdon Life Sciences (HLS, now Envigo), the third largest contract toxicology company in the world. He specialized in practical improvement of the quality and efficiency of test methods, many of which are still in routine use including Ames, in vivo micronucleus, in vitro and in vivo chromosome aberration. While at Huntingdon, he was awarded a master's degree for work on in vitro and in vivo UDS by Swansea University, which remains the academic center of excellence for genetox in the UK.

After 21 years, he left HLS as a principal toxicologist responsible for all in vivo genetox to move to Charles River Laboratories (CRL) in Montreal, Quebec, the largest toxicology facility in the world. At Montreal, he designed the laboratories then established and managed the new Genetic Toxicology facility. He trained the staff and helped set up and validate all the current standard assays from Ames to in vivo comet. The department soon established a sound reputation for quality and on-time reporting in a minimum time-frame while achieving rapid growth and good profitability. He was also responsible for heading the in vitro and in vivo drug metabolism department for a temporary (6-month) period during this time. Ray left CRL after 11 years and has since worked as a Principal Toxicologist at Bausch and Lomb (specializing in ophthalmic drugs and medical devices), as Scientific Director at Moltox (responsible for genetic toxicology testing product improvement and development) and as an independent consultant in genetic toxicology.

He has authored or co-authored numerous technical papers on genetic toxicology testing and was directly involved in the international validation of the rat micronucleus test and its integration into longer-term toxicology tests. During his career, he has trained many technical and scientific staff, written and reviewed many thousands of genetox reports and has directed innumerable studies intended for regulatory submission under European, Japanese, North American and OECD GLPs. He has worked on all sorts of materials including gases, diesel exhaust, dyes and pharmaceutical impurities. He plays squash well and golf badly.

Affiliations and expertise

Boone, North Carolina, USA

Life Sciences

Physical Sciences & Engineering

Social Sciences & Humanities

Health