Hair analysis drug metabolites

Figure 3.1 A hair sample from a suspected drug user is prepared for forensic analysis. As hair grows, it incorporates small amounts of chemicals that are produced when drugs are broken down in the body. To identify these drugs, the hair is first cut into pieces and soaked in a liquid solvent The solvent removes the traces of drug metabolites from the hair so that they can be identified by chromatography and mass spectrometry.

Lew authors described antidepressant analysis in alternative specimens, such as hair or oral fluid. LC-CID-MS and MS/MS mass spectra libraries for identification of several drugs were employed by Muller et al. [32] for the detection of maprotiline, citalopram, and their desmethyl metabolites in authentic hair specimens extracted ions chromatograms were employed for subsequent antidepressant quantification. Also Klys et al. [33] applied LC-MS/MS to the analysis of blood, urine, and hair specimens in a fatal case due to clomipramine overdose in combination with alcohol. Blood clomipramine and norclomipramine concentrations explained the fatal outcome, and hair analysis confirmed that the deceased was on clomipramine treatment for, at least, 12 months prior to his death. With regard to oral fluid analysis, de Castro et al. [34] developed and validated a... 

Yes. In situations where drugs are known to be present in the environment, it is easy to demonstrate that passive exposure can produce positive hair analysis results. In a study by Haley and Hoffmaim of the nicotine and cotinine concentrations in the hair of smokers and nonsmokers, there appeared to be a higher average of nicotine in the unwashed hair of smokers (average 8.75 ng/mg). However, nonsmokers also had an appreciable level of nicotine (average 2.42 ng/mg) which overlapped that of smokers. In contrast, cotinine (the nicotine metabolite) does appear to be a marker of tobacco use in this population. More recently, Kintz and co-workers and Kintz proposed a cutoff level of 2 ng of nicotine/mg of hair to ehminate nonsmoking individuals. Even at this level, some nonsmokers would be positive. 

The evidence from this laboratory and from others reviewed in this chapter reinforces and extends the serious concern that external contamination of hair by drugs of abuse can easily occur. Any interpretation of hair analysis data should consider the prospect that the sample could have been externally contaminated. The pharmacokinetics of the incorporation of drugs into many tissues has been well elaborated. However, substantial additional information on the mechanisms for incorporation of drugs into hair, the decontamination of hair, the differentiation between exposure to exogenous and endogenous drugs, and the meaning of the presence of metabolites in the hair are needed before hair analysis can be employed in many forensic applications. 

Springfield, A. C., CartmeU, L. W., Aufderheide, A. C., Buikstra, J., and Ho, J., Cocaine and metabolites in the hair of ancient Peruvian coca leaf chewers. Forensic Sci. Int., 63, 269, 1993. Baumgartner, W. A. and HiU, V. A., Sample preparation techniques. Forensic Sci. Int., 63,121,1993. Nakahara, Y. and Kikura, R., Hair analysis for drugs of abuse. VII. The incorporation rates of cocaine, benzoylecgonine cmd ecgonine methyl ester into rat hair and hydrolysis of cocaine in rat hair. Arch. Toxicol, 68, 54,1994. 

Due to the low amount of specimen used for hair analysis and the low concentration of drugs and their metabolites in hair, extraction procedures are mostly necessary to concentrate the analytes in the tested solution. Various methods have been used for this step. The variety of described procedures show that there is no universal solution. The choice of an extraction procedure may be influenced by the kind of drug and drug metabolite(s) to be analyzed and the method used for the subsequent analysis. Preceding the real extraction, there are four treatments with more or less intermethodical differences to prepare the hair matrix for the extraction ... 

GC/MS Analysis for Miscellaneous Drugs/Drug Metabolites in Hair... 

Independent from this controversy, hair analysis for drugs offers a way to uncover chronic use in cases where blood or urine analyses fail. The detection of the parent drug and the metabolites seem to be a possibility to decide whether the drug has been incorporated or not. In cases of incorporation, the concentration of the more lipophilic parent drug seems to be nearly always higher than that of the metabolites, ° ° contrary to urinalysis. 

Nakahara, Y., Ochiai, T., Kikura, R., Hair analysis for drugs of abuse. V. The facility in incorporation of cocaine into hair over its major metabolites, benzoylecgonine and ecgonine methyl ester. Arch. Toxicol, 66, 446-449, 1992. 

The growing concern regarding the widespread abuse of illicit drugs has inspired the development of new technologies for the analysis of therapeutic and abused drugs and their metabolites in unusual biological tissues, including hair. Analysis of... 

In a study of maternal drug use by Kintz and Mangin, hair samples were collected from neonates with confirmed in utero exposure to heroin. Morphine was present in all samples in a concentration range similar to previous reports based on analysis of hair from adults. Codeine was also present in aU samples, but in low concentration. Maternal hair was not tested. Urine samples were also collected and tested for opioids. Four (44%) samples were positive for morphine and 6-AM. The data indicate that drug and/or drug metabolite is transferred to the fetus through the placenta and retained in hair. 

Goldberger, B. A., Darraj, A. G., Cone, E. ]., and Caplan, Y. H., Detection of methadone, methadone metabolites, and other illicit drugs of abuse in hair of methadone treatment subjects, 2nd International Meeting on Clinical and Forensic Aspects of Hair Analysis, Genova, Italy, 1994. 

For the accurate interpretation of a positive hair analysis result, it is necessary to subject hair to specially developed wash and extraction procedures. During the development of these procedures, we discovered several important properties of hair, properties which were essential for defining three unique wash kinetic criteria and their cutoff levels which, in conjunction with washing, enhance the acciuacy of the interpretation of a positive hair analysis result. Additional certainty can be achieved by the measurement of metabolites or metabolite/drug ratios, by the development of ultrasensitive gas chromatography/tandem mass spectrometry (GC/MS/MS) or ion... 

Overall, hair analysis provides convincing evidence of past exposure to a drug. Viala et al. identified by GC/MS chloroquine and its major metabolite in hair samples of patients who received the antimalaria drug for several months. 

The nonplant synchrotron IR microspectroscopic analysis at the NSLS beamline included drug metabolites in hair [31], depth profiling of photodecomposition of polymer layers [19], and numerous mammalian tissue probings, including the brain tissue of rats that had consumed D2O in their drinking water [10]. A summary of the plant material experience from BNL over a continuous 15-month period was reported in 1998 [17], and included the spectra of individual cells within a wheat primary root and the mapping of transitions between the botanical parts of wheat, safflower, oats, corn and barley. 

As with urine, saliva (spumm) is easy to collect. The levels of protein and lipids in saliva or spumm are low (compared to blood samples). These matrices are viscous, which is why extraction efficiency of xenobioties amoimts to only 5 to 9%. By acidifying the samples, extraction efficiencies are improved as the samples are clarified, and proteinaceous material and cellular debris are precipitated and removed. Some xenobioties and their metabohtes are expressed in hair. Hair is an ideal matrix for extraction of analytes to nonpolar phases, especially when the parent xenobioties are extensively metabolized and often nondetectable in other tissues (parent molecules of xenobioties are usually less polar than metabolites). Hair is a popular target for forensic purposes and to monitor drug compliance and abuse. Human milk may be an indicator of exposure of a newborn to compounds to which the mother has been previously exposed. The main components of human milk are water (88%), proteins (3%), lipids (3%), and carbohydrates in the form of lactose (6%). At present, increasing attention is devoted to the determination of xenobioties in breath. This matrix, however, contains only volatile substances, whose analysis is not related to PLC applications. 

Results of hair tests for abused drugs such as cocaine and opioids can provide useful information regarding drug use. Compared to urine analysis, hair testing provides an extended window of analyte (parent and metabolites) detection which cannot be affected by short periods of abstinence. In addition, collection of hair is considered by most a noninvasive technique, and retesting through a second sample is possible. Also, in contrast to urine, hair is not readily adulterated. 

Moriya, R, Miyaishi, S., and Ishizu, H., Presumption of an history of methamphetamine abuse by postmortem analyses of hair and nails a case report, Jpn. ]. Alcohol Drug Depend., 27,152,1992. Nakahara, Y., Kikura, R., Takahashi, K., and Kunoma, K., GC/MS analysis of drugs and metabolites in hair for diagnosis of chronic methamphetamine abuse, Adv. Chem. Diagnos. Treat. Metab. Disord., Matsumoto, Ed., London, 1994,187. 

So from a simple mass transport point of view, hair is less susceptible to contamination by exogenous mechanisms than urine by endogenous mechanisms. But then hair, but not urine, is cleansed by normal hygienic practices and by special laboratory wash procedures. The latter are further strengthened by special kinetic analysis of the wash data. And, finally, the deposition of exogenous drugs onto hair, unlike their accumulation in urine, is not associated with the formation of metabolites. 

The ultimate aim in the analysis of hair in relation to DFCs, is the detection of a single dose of a drug. It is also desirable to detect metabolites of the drug to verify that the source of the drug in the hair is from the body and not from the environment. The low concentrations encountered, in the low pg/mg range, dictates the use of highly sensitive detection systems. The amazing developments in LC and tandem mass spectrometry in recent years make this combination of techniques ideal for the task. 

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