Drugs Of Abuse In Hair Research Articles

Detection of methadone, methadone metabolites, and other illicit drugs of abuse in hair of methadone-treatment subjects.

The disposition of methadone, its metabolites, and other illicit drugs of abuse was investigated in head hair samples collected from heroin users (N = 20) enrolled in an outpatient detoxification study. Hair samples were assayed for methadone, methadone primary metabolite (2-ethylidene-1,5-dimethyl-3,3-diphenylpyrrolidine, EDDP), methadone secondary metabolite (2-ethyl-5-methyl-3,3-diphenylpyrroline, EMDP), cocaine, phencyclidine, heroin, and 6-acetylmorphine. Hair samples were cut, washed, and incubated in methanol. The methanolic hair wash and incubation fractions were purified with solid-phase extraction and assayed by gas chromatography-mass spectrometry. Methadone, methadone metabolites, cocaine, and phencyclidine were assayed quantitatively, and other drugs were measured qualitatively. The number of positive results and the corresponding concentration ranges were as follows: methadone, 0-15.0 ng/mg (N = 18); EDDP, trace (N = 13); EMDP, trace (N = 1); cocaine, 0->40 ng/mg (N = 14); phencyclidine, 0-1.5 ng/mg (N = 2); heroin, positive (N = 3); and 6-acetylmorphine, positive (N = 4). These data suggest that testing hair for methadone, methadone metabolites, and other illicit drugs of abuse may be useful to drug-treatment specialists as a means of verifying drug-use history, monitoring compliance, and providing a broad measure of drug exposure.

Interlaboratory comparison of quantitative determination of amphetamine and related compounds in hair samples

Testing human hair for drugs of abuse is a relatively new technique which requires control before being fully accepted in justice applications. Laboratories must be able to demonstrate that they can accurately determine what drugs are present in unknown hair samples and at what levels. To date few exercises have been organized in USA, Germany and France, all devoted to opiates, cocaine and cannabis. However, the number of drugs which can be detected in hair is growing every day. Among them, amphetamine and related compounds, such as MDMA, are of major interest due to increasing abuse. At the initial stage of this work, four different preparation procedures were used to test amphetamine, MDA and MDMA. Direct methanol extraction, acid (HCl 0.1 N), alkaline (NaOH 1 N) and enzymatic (β-glucuronidase/arylsulfatase) hydrolyses were compared. Best recoveries were observed after alkaline hydrolysis. The same hair sample was powdered and sent to 16 laboratories, in USA (4), Germany (6), France (3), Spain (1), Japan (1) and Korea (1) to test amphetamine, methamphetamine, MDA and MDMA. All laboratories returned results within 3 months. Amphetamine tested positive 13 times with concentrations ranging from 3.3 to 17.5 ng/mg. Only 2 laboratories identified methamphetamine, using GC/MS, at low concentration (0.8 and 1.8 ng/mg), which appears to be a false positive. MDA and MDMA both tested positive in 14 cases, with concentrations ranging from 1.8 to 19.5, and 8.9 to 100.0 ng/mg for MDA and MDMA, respectively. These scattered results clearly indicated that new exercises are needed to ensure quality in hair testing. This is one of the major aims of the Society of Hair Testing.

Hair analysis as evidence in forensic cases.

The analysis of hair for drugs of abuse is a powerful tool useful in answering questions that cannot be solved by the detection of drugs in body fluids, e.g., blood, urine, or saliva. The most frequent forensic investigations occur in cases where narcotics laws are offended and in questions of criminal responsibility where the chronic use of a drug is an issue. In the list of drugs of abuse, heroin, cocaine, amphetamine, and cannabis are the drugs of abuse that are most frequently involved in judicial inquiries.

Interlaboratory studies on the analysis of hair for drugs of abuse: results from the fourth exercise.

Fourteen laboratories interested in the analysis of human hair for drugs of abuse participated in a fourth interlaboratory study to determine how well drugs could be detected and quantitated in hair. The drugs of abuse included cocaine, benzoylecgonine, cocaethylene, codeine, morphine, and 6-monoacetylmorphine. The hair samples analyzed were in the form of short segments and included hair from drug users, soaked hair (drug-free hair into which drugs had been soaked), and drug-free hair. Results from the study show that the laboratories performed well qualitatively, but that there was a large amount of scatter in quantitative results. Various methods were used to extract the drugs from the hair, and the most commonly used approaches, HCl extractions, methanol extractions, and enzyme digestions, all gave comparable results. Gas chromatography-mass spectrometry (GC-MS) was the most commonly used analytical technique. Of the laboratories using GC-MS, some produced consistently good results, whereas others produced results of poorer quality. Two laboratories using liquid chromatography-mass spectrometry produced good results, but the one laboratory using liquid chromatography without mass spectrometric detection produced less accurate results. Laboratories that reported good results on control material sent with the unknowns generally performed better on the unknowns than did laboratories whose results on the control were less accurate.

Determination of Fentanyl in Hair: The Case of the Crooked Criminalist

When a State Crime Laboratory Director found that fentanyl patches were missing from a case submission stored in his evidence vault, he performed an investigation of those members of his staff who had access to the materials. As part of this investigation, two staff members were forced to submit to hair testing for fentanyl and other opiates and opioids. This unusual testing protocol was used to identify a senior Criminalist as a chronic abuser of fentanyl, and led to the rapid resolution of the case. To the best of our knowledge, this is the first report of the forensic analysis of hair for drugs of abuse in which the technology was used to identify an individual as a chronic fentanyl abuser.

Interlaboratory comparison of quantitative determinations of drug in hair samples

Testing human hair for drugs of abuse is a relatively new technique which requires control before being fully accepted in Justice applications. A consensus procedure was recently proposed to the four French Laboratories performing hair analysis for opiates and cocaine. Results of two independent controls have shown that the laboratories have performed very well quantitatively, using the recommended method. In order to compare these results with those obtained by other procedures, one sample was sent to 15 laboratories concerned with the analysis of human hair for drugs of abuse in Germany, Italy, Spain, and United States. Results from this study have indicated that the French recommended method is in accordance with the general procedures.

Decontamination procedures for drugs of abuse in hair: are they sufficient?

This paper reviews the methods for decontaminating hair exposed to external solutions of drugs of abuse. Exposure of hair to cocaine at 1 μg/ml for 5 min is sufficient to contaminate hair, yet decontamination is a very slow process. Using externally contaminated hair, a number of decontamination procedures were attempted, and none removed all the contamination. The percentage of external contamination removed depended on the hair type, with thick black hair being the most resistant to decontamination. Hair treated by dying incorporated externally applied drugs differently, depending on the hair type. Thick black hair became more absorbent whereas thin brown hair became less absorbent. Kinetic wash criteria are evaluated for their ability/inability to determine if hair has been contaminated from external sources. A theoretical framework for the incorporation and removal of drugs from hair is discussed, and the hypothesis that inaccessible domains exist in hair which trap drugs is critically examined. The results presented in this paper strongly suggest that much more information on the decontamination of hair and the differentiation of exogenously and endogenously incorporated drugs is needed before hair analysis can be employed in most forensic applications. We propose that the radioactive tracer methods discussed herein are well suited for evaluating any new decontamination or extraction technique.

Capillary electrophoresis: a new tool in forensic toxicology. Applications and prospects in hair analysis for illicit drugs

Capillary electrophoresis, the modern approach to instrumental electrophoresis, is probably the most rapidly expanding analytical technique that has appeared in recent years. In the hands of forensic toxicologists, capillary electrophoresis (CE) represents a powerful new analytical tool, which has proved suitable for the investigation of illicit drugs in seized preparations and also in complex biological matrices, among which is hair. CE can be applied according to different separation mechanisms, and among those that are toxicologically relevant are capillary zone electrophoresis and micellar electrokinetic capillary chromatography, which display different selectivities. For the investigation of hair for drugs of abuse, capillary electrophoresis proved effective, providing simultaneous determinations of different drugs without derivatization, with acceptable sensitivity (typically better than 1 ng of drug per mg of hair). The possibility of carrying out determinations of the same analytes, based on different separation mechanisms (capillary zone electrophoresis and micellar electrokinetic chromatography) with the same instrumentation, simply changing the buffer composition, provides an interesting possibility of ‘internal’ confirmation of the results.

NIST reference materials to support accuracy in drug testing

Substance abuse is a major problem worldwide. There is considerable emphasis placed upon testing individuals for evidence of use of controlled substances. Because the consequences of a positive test can be quite severe, laboratories conducting such tests must rigorously follow a carefully designed quality assurance program. Such a QA program should include use of reference materials to assure that the methods used to detect and quantify drugs are providing accurate results. The National Institute of Standards and Technology (NIST) supports accuracy in drugs of abuse testing by providing Standard Reference Materials (SRMs) with certified concentrations of drugs of abuse in urine- and hair-based reference materials. NIST, working in collaboration with the College of American Pathologists (CAP), has developed urine-based SRMs for marijuana metabolite, cocaine metabolite, morphine and codeine, and morphine glucuronide and CAP Reference Materials for amphetamines and phencyclidine. Certification measurements performed at NIST involve two independent methods for each analyte, one of which always uses GC/MS with the other usually being an LC method with either MS or UV detection. Work has recently been completed on a seven component drug in urine SRM. In addition NIST conducts research in the analysis of hair for drugs of abuse. To assist laboratories testing hair for that purpose, NIST has developed two drugs in hair reference materials.

Testing human hair for drugs of abuse. IV. Environmental cocaine contamination and washing effects

Active cocaine use results in sequestration of parent drug in hair. In addition, hair has unique physicochemical properties that permit absorption of cocaine from the environment. When hair is tested for evidence of cocaine, it is important to consider whether the positive test resulted from active drug use or environmental contamination. In a series of laboratory experiments, it was found that exposure of ‘cut’ hair to cocaine vapor (‘crack’ smoke) and to aqueous solutions of cocaine hydrochloride resulted in significant contamination of hair samples. Similar results were obtained with two subjects who were exposed to cocaine vapor in an unventilated room. The amount of contamination adsorbed by hair depended upon both time and extent of exposure. Washing the hair samples with methanol removed >70% of the cocaine contaminant after cocaine vapor exposure, but was less effective (<50%) following contamination with aqueous cocaine. Shampoo treatment cycles (overnight soaking) progressively removed increasing amounts of cocaine from the contaminated hair, but residual cocaine remained after 10 cycles. Studies were also performed to determine the usefulness of benzoylecgonine as a marker of active cocaine administration. Small amounts of benzoylecgonine (ca. 1 ng/mg) were formed in hair as a result of environmental contamination with cocaine. Also, it was found that benzoylecgonine could be adsorbed from illicit cocaine contaminated with benzoylecgonine. It was concluded that positive hair test results should be interpreted cautiously due to the possibility of environmental contamination from cocaine and related constituents.

Radioimmunoassay of drugs of abuse in hair. Part 1: Methadone in human hair, method adaptation and the evaluation of decontamination procedures

A method suitable for the determination of methadone in human hair is presented. Adaptation and evaluation of a solid-phase I125 radioimmunoassay, designed for the quantitative measurement of methadone in urine, and development of a pre-analytical wash procedure has enabled a specific, sensitive and accurate analytical procedure to be developed. The specificity of the antiserum towards other drugs or biologically active compounds is evaluated up to a concentration of 100,000 ng ml-1 and accuracy covering a range of 0-450 ng ml-1 is found to be within 6% of expected methadone concentrations prepared in both drug free hair extract and urine. Inter-assay relative standard deviation (RSD) at concentrations of 5.1, 76.0 and 247 ng ml-1 methadone are 5.5, 2.5 and 3.6% respectively (n = 10) and intra-assay RSD at concentrations 2.3, 25.2 and 217 ng ml-1 are 5.3, 3.6 and 6.8% (n = 5). The limit of detection is 0.5 ng ml-1. Extraction of control drug free hair samples spiked with methadone at concentrations of 100, 250 and 400 ng ml-1 achieved recoveries of 86, 80 and 89%, respectively. Control hair samples contaminated with methadone are examined under differing wash procedures to assess their effectiveness in the removal of methadone contaminant. A suitable pre-analytical wash regime is proposed for removal of contaminant derived from external or environmental sources. The mechanics of the wash action and contaminant application to the hair is discussed. It is concluded that the adapted radioimmunoassay and developed pre-analytical decontamination procedure is a suitable technique to employ for the measurement of methadone in human hair, be it prescribed or abused, with concentrations expressed as ng methadone per mg hair.

Interlaboratory comparison studies on the analysis of hair for drugs of abuse

Eleven laboratories interested in the analysis of human hair for drugs of abuse participated in a study to determine how well drugs could be detected and quantified in hair. For the two exercises completed to date, substances to be determined were limited to cocaine, benzoylecgonine, and morphine. Samples sent to the participating laboratories included hair from drug users, drug-free hair, and hair into which drugs had been soaked. For the first exercise, the hair samples were sent as powders; for the second, they were in the form of short segments. Results from these studies have shown that the laboratories, with a few exceptions, have performed very well qualitatively. However, scatter in quantitative results was high. Various approaches were used to liberate drugs from the hair, with the most commonly used, acid extractions and enzyme digestions, producing similar results. Laboratories using GC MS generally performed well and reported no false positives. In contrast, one laboratory analyzing hair directly using MS MS without extractions produced three of the four false positives and the worst quantitative results.

Ion trap mass spectrometry, a new tool in the investigation of drugs of abuse in hair

The use of an ion trap mass spectrometer for the detection, via tandem mass spectrometry, of morphine and cocaine in the hair of drug addicts proved to be successful. In particular, detection limits of 1–20 ppb for morphine and 5–30 ppb for cocaine were achieved, with good reproducibility and absence of interferent species.

The occurrence of cocaine, heroin and metabolites in hair of drug abusers

The analysis of hair for drugs of abuse reveals information regarding past drug exposure. We developed methods for washing, extraction and analysis of hair samples for cocaine, heroin and metabolites. Twenty paired head- and arm-hair samples, collected from known heroin/cocaine abusers, were analyzed with a new comprehensive GC MS assay for cocaine, heroin and metabolites. Cocaine and 6-acetylmorphine (6-AM) were the major analytes present in both head- and arm-hair samples. Cocaine was detected in all head- and 17 arm-hair samples. The concentration of cocaine found was 4–760 ng 10 mg in head hair and 0–1090 ng 10 mg in arm hair. Less benzoylecgonine was present in a concentration range of 0–158 ng 10 mg of head hair and 0–125 ng 10 mg of arm hair. Heroin was found in only 2 head-hair samples, whereas 6-AM was present in 14 head and 6 arm-hair samples. The concentration of 6-AM was 0–8 ng 10 mg in head hair and 0–31 ng 10 mg in arm hair. Morphine was present in 3 head-hair samples in a range of 2–9 ng 10 mg and was not detected in arm-hair samples. When results were compared by groups (head hair versus arm hair, Caucasoid versus Africoid), only two significant differences were found. Cocaine concentrations in both head and arm hair were significantly ( P < 0.05) higher in the Africoid group than in the Caucasoid group. The reasons for these differences were not readily apparent, but could have been due to differences in the level of cocaine use or to ethnic differences in the deposition of drug in hair.

Testing Human Hair for Drugs of Abuse. III. Identification of Heroin and 6-Acetylmorphine as Indicators of Heroin Use

Hair samples from 20 documented heroin users contained 6-acetylmorphine, a unique metabolite of heroin, in all samples. Heroin was identified in smaller amounts in seven of these samples. The identity of 6-acetylmorphine and heroin was established by comparison of full scan spectra of extracts to standard reference materials. The presence of 6-acetylmorphine generally predominated over heroin, morphine, and codeine. The mean concentrations of analytes were as follows: 6-acetylmorphine, 0.90 ng/mg, N = 20; heroin, 0.17 ng/mg, N = 7; morphine, 0.26 ng/mg, N = 20; codeine, 0.18 ng/mg, N = 15. Analysis of hair samples obtained from 10 drug-free control subjects were negative for 6-acetylmorphine, morphine, and codeine. However, a small interfering peak was observed at the retention time for heroin. Control samples soaked in aqueous solutions of heroin and 6-acetylmorphine were found to be contaminated, even though an initial wash step was included in the analysis. These data suggest that hair analysis for 6-acetylmorphine can be used to differentiate heroin users from other types of opiate exposure (e.g., poppy seed, licit morphine, and codeine); however, environmental contamination can potentially produce false positives during opiate testing.

Testing Human Hair for Drugs of Abuse. II. Identification of Unique Cocaine Metabolites in Hair of Drug Abusers and Evaluation of Decontamination Procedures

Two unique metabolites of cocaine, cocaethylene and norcocaine, were identified by GC/MS in the hair of cocaine users. Their presence cannot be explained by environmental contamination; thus, their presence together with cocaine provides convincing evidence that cocaine is excreted in hair after active cocaine administration. The amount of cocaine in hair predominated over all metabolites generally by a factor of 5-10. Two washing procedures were evaluated for their efficiency in removal of cocaine from environmentally contaminated hair. Neither procedure completely removed cocaine, suggesting that false positives can result from environmental contamination. Analysis of the methanolic wash of the hair of cocaine users also revealed the presence of cocaine metabolites, indicating that washing removes cocaine from the interior as well as from the exterior surface of hair during decontamination procedures.

Testing Human Hair for Drugs of Abuse. I. Individual Dose and Time Profiles of Morphine and Codeine in Plasma, Saliva, Urine, and Beard Compared to Drug-Induced Effects on Pupils and Behavior*

The time course of appearance of morphine and codeine in beard after single dose administration in two human subjects was monitored by radioimmunoassay and confirmed by gas chromatography/mass spectrometry. Both morphine and codeine appeared in beard approximately 7-8 days after drug administration at a time when drug levels in urine, plasma, and saliva were not detectable and drug-induced effects had disappeared. Drug levels in beard appeared to be dose-related suggesting that hair analysis can provide evidence of time and degree of drug exposure.