Gc/Ms Assays for Abused Drugs in Body Fluids

GC/MS ASSAYS FOR ABUSED DRUGS IN BODY FLUIDS U.S. DEPARTMENT OF HEALTH AND HUMAN SERVICES • Public Health Service • Alchol, Drug Abuse, and Mental Health Administration GC/MS Assays for Abused Drugs in Body Fluids Rodger L. Foltz, Ph.D. Center for Human Toxicology University of Utah Salt Lake City, Utah 64112 Allison F. Fentiman, Jr., Ph.D. Ruth B. Foltz Battelle Columbus Laboratories Columbus, Ohio 43201 NIDA Research Monograph 32 August 1980 DEPARTMENT OF HEALTH AND HUMAN SERVICES Public Health Service Alcohol, Drug Abuse, and Mental Health Administration National Institute on Drug Abuse Division of Research 5600 Fishers Lane Rockville, Maryland 20857 For sale by the Superintendent of Documents, U.S. Government Printing Office Washington, D.C. 20402 The NIDA Research Monograph series is prepared by the Division of Research of the National Institute on Drug Abuse. Its primary objective is to provide critical reviews of research problem areas and techniques, the content of state-of-the- art conferences, integrative research reviews and significant original research. Its dual publication emphasis is rapid and targeted dissemination to the scientific and professional community. Editorial Advisory Board Avram Goldstein, M.D. Addiction Research Foundation Palo Alto, California Jerome Jaffe, M.D. College of Physicians and Surgeons Columbia University, New York Reese T. Jones, M.D. Langley Porter Neuropsychiatric Institute University of California San Francisco, California William McGlothlin, Ph.D. Department of Psychology, UCLA Los Angeles, California Jack Mendelson, M.D. Alchol and Drug Abuse Research Center Harvard Medical School Mclean Hospital Belmont, Massachusetts Helen Nowlis, Ph.D. Office of Drug Education, DHHS Washington, D.C. Lee Robins, Ph.D. Washington University School of Medicine St. Louis, Missouri NIDA Research Monograph series William Pollin, M.D. DIRECTOR, NIDA Marvin Snyder, Ph.D. DIRECTOR, DIVISION OF RESEARCH, NIDA Robert C. Petersen, Ph.D. EDITOR-IN-CHIEF Eleanor W. Waldrop MANAGING EDITOR Parklawn Building, 5600 Fishers Lane, Rockville, Maryland 20857 GC/MS Assays for Abused Drugs in Body Fluids ACKNOWLEDGMENTS Support for the development and evaluation of the GC/MS assays described in this monograph and the preparation of the manuscript was provided by the National Institute on Drug Abuse, contracts ADM 45-74-140, HSM 42-72-183, and 271-77-3419 to Battelle Columbus Laboratories, and contract 271-80-3719 to the Center for Human Toxicology, and by the Insurance Institute for Highway Safety, proj- ect 6762 at the Center for Human Toxicology. The United States Government does not endorse or favor any specific commercial product, commodity, or supplier. Trade or proprietary names and names of suppliers appearing in this publication are used only because they are considered essential in the context of the information presented. Library of Congress catalog card number 80-600143 DHHS publication number (ADM) 80-1014 Printed 1980 NIDA Research Monographs are indexed in the Index Medicus. They are selectively included in the coverage of Biosciences Information Service, Chemical Abstracts, Current Contents, Psychological Abstracts, and Psychopharmocology abstracts. iv Foreword This monograph presents a collection of methods for the quantitative analysis of several important drugs of abuse by the technique of gas chromatography-mass spectrometry (GC/MS). The material is arranged textbook fashion, by chapters on specific drugs, to assist researchers in setting up GC/MS assay procedures in their own laboratories. These assays are particularly suited to basic studies on the pharmacology and pharmacokinetics of drugs of abuse. Forensic toxicology, which sometimes requires the highly sensitive and specific sample analysis provided by GC/MS, is another important area of application. Even when an alternate method such as immu- noassay is available for such analyses, GC/MS is often the only means of providing a confirmation assay. The sensitivity and specificity of GC/MS are especially critical to studies on the impact of drugs on traffic safety. In addition, demand for leg- islation relating to drugs and highway safety is increasing, and enforcement of such laws will necessitate appropriate sample assays. The unique characteristics of the GC/MS analytical method make it the only means of determining drug levels under certain circum- stances. The inherent complexity of the technique, however, often dissuades investigators from embarking on studies that involve its use. At a time when basic questions underlying drug abuse call for increasingly sophisticated methods of data acquisition, this monograph should provide the information needed to make one of these procedures easier to establish and to encourage new areas of research. Marvin Snyder, Ph. D. Director, Division of Research National Institute on Drug Abuse v Preface Soon after the establishment of the National Institute on Drug Abuse in 1974, the development of an analytical program empha- sizing gas chromatography/mass spectrometry (GC/MS) was initiated. A major part of this program was directed at the development, testing, and eventual publication of GC/MS methods suitable for a number of abused drugs of interest to this Institute. It was felt that by making available carefully described quantitative procedures for these drugs, more researchers would be encouraged to move into several critically important areas of investigation. A major segment of this development and testing program was carried out by Battelle (Columbus Laboratories and at the Center for Human Toxicology at the University of Utah. These GC/MS methods empha- size chemical ionization techniques which frequently offer significant advantages in sensitivity and specificity over techniques of electron impact ionization. Each of the techniques described in this monograph has been field tested by one or more independent laboratories in order to optimize both the procedure itself and the clarity of its presentation. This field testing was carried out by Battelle and consisted of sending the participating laboratories a series of known standard samples along with the information brochures included herein as chapters. The participating laboratories then set up the method in their own facilities and analyzed the spiked samples. Problems with either the instructional material or the method allowed Battelle to make many improvements to insure the highest probability of an inexperienced lab’s success with one of these assays. This monograph represents the combined efforts of many individuals and has evolved to its present state over several years. We hope that it will save research investigators valuable time in estab- lishing quantitative essays of these drugs in their own laboratories. The basic principles of GC/MS quantitative analysis as presented here should be useful in setting up procedures for other drugs as well. Richard L. Hawks , Ph.D. Division of Research vii Contents Foreword . v Marvin Snyder Preface . vii Richard L. Hawks Chapter 1 Introduction . 1 Chapter 2 Experimental Considerations and Operations Common to All of the Assays . 5 Chapter 3 Phencyclidine(PCP) . .25 Chapter 4 Methaqualone . .39 Chapter 5 Methadone . .51 Chapter 6 -Tetrahydrocannabinol (THC) and Two of Its Metabolites, 11-Hydroxy- -THC and 11-Nor-9-Carboxy- -THC . .62 Chapter 7 Cocaine and Its Major Metabolite, Benzoylecgonine . 90 Chapter 8 Morphine . , . 110 Chapter 9 Diazepam and Its Major Metabolite, N-Desmethyl- diazepam . 128 Chapter 10 Amphetamine . 150 Chapter 11 Methamphetamine . 165 Chapter 12 2,5-Dimethoxy-4-Methylamphetamine (DOM) . 179 Chapter 13 Mescaline . , . 189 ix Chapter 1 Introduction The combination of gas chromatography and mass spectrometry (GC/MS) became widely recognized in the 1960’s as the most sensitive and versatile tool available for the identification of volatile organic compounds. It was not until the 1970’s, however, and the development of selected ion monitoring techniques, that the poten- tial of GC/MS for quantitative analysis was generally appreciated. It now seems certain that the quantitative measurement of specific organic compounds in complex mixtures will become the major application of GC/MS simply because its sensitivity, accuracy, and ver- satility are unequalled by any other technique, including the popu- lar immunological assays. The rapid increase in the use of GC/MS has occurred in spite of several severe limitations and deterrents. The major limitation is that for a compound to be analyzed by GC/MS it must have suf- ficient volatility and thermostability to pass through the gas chro- matographic column intact in the vapor state, or be capable of con- version to a derivative which can do so; this requirement prohibits GC/MS analysis for 80 to 90 percent of known organic compounds. Fortunately, this is not a serious problem for the toxicologist, because most drugs and their metabolites are among the structures which can be analyzed by GC/MS. More serious deterrents to wider use of GC/MS in toxicological and clinical laboratories are the expense of GC/MS instrumentation and the relatively high level of training and skill required to make effective use of these instru- ments. Although the former is unlikely to be reduced in the fu- ture, the latter will undoubtedly become less burdensome as the reliability of GC/MS instrumentation is improved and its design simplified. The purpose of this monograph is to assemble, within a single vol- ume, detailed descriptions of GC/MS

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