Chapter 28

Hair in Forensic Medicine

Bianca Maria Piraccini, Massimiliano Pazzaglia, Antonella Tosti

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Synonyms hair drug screening test, forensic hair analysis

Key Features

Drugs, chemicals, and biological substances are

stored in hair where they can be detected and measured. In hair samplings, the window of drug detection extends to months or even years. Collection is noninvasive, relatively easy to perform, and in forensic situations it may be achieved under close supervision to prevent adulteration or substitution.

The sample size required for analysis is small and is easily stored at room temperature. Possible applications of hair analysis include diag-

nosis of drug abuse, poisoning and doping, evaluation of occupational exposure to toxics, evaluation of prenatal exposure to drugs, and monitoring of a patients compliance with drug prescription.

Contents 28.1 28.2 28.3 28.4 28.4.1 28.4.2 28.5 28.6 28.6.1 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . History . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Pathophysiology of Incorporation of Substances into Hair . . . . . . . . . . . . . . . . Techniques of Analysis [8, 9] . . . . . . . . . . . Hair Sampling . . . . . . . . . . . . . . . . . . . . . . . Decontamination, Extraction, and Detection . . . . . . . . . . . . . . . . . . . . . . . Clinical Interpretation . . . . . . . . . . . . . . . . Purposes of Hair Analysis . . . . . . . . . . . . . Forensic Investigations . . . . . . . . . . . . . . . . 539 540 540 541 541 541 541 541 541 28.6.1.1 28.6.1.2 28.6.2 28.6.3 28.6.4 28.6.5 28.7 Drug Abuse and Poisoning . . . . . . . . . . . . Rape Cases . . . . . . . . . . . . . . . . . . . . . . . . . . Doping Controls . . . . . . . . . . . . . . . . . . . . . Occupational Medicine . . . . . . . . . . . . . . . Prenatal Exposure . . . . . . . . . . . . . . . . . . . . Patients Compliance to Drug Prescription . . . . . . . . . . . . . . . . . . . . . . . . . Future Developments . . . . . . . . . . . . . . . . . Summary for the Clinician . . . . . . . . . . . . 541 541 541 541 542 542 542 542

References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 542

28.1

Introduction

Hair analysis for drugs and toxics has recently become very popular in several fields of medicine, including forensic and clinical toxicology and occupational medicine [2, 4, 8, 9] (Table 28.1). Hair may in fact give a long-term history of drug intake and abuse as well as toxin exposure and therefore

represents an unique substrate for forensic purposes. Collection is noninvasive, relatively easy to perform, and in forensic situations it may be achieved under the close supervision of law enforcement officers to prevent adulteration or substitution. The sample size required for analysis is small and is easily stored at room temperature. The window of drug detection dramatically extends to months or even years, because long scalp hair may

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B. M. Piraccini, M. Pazzaglia, A. Tosti Table 28.1 Drugs, chemical, and biological substances that can be detected and measured in hair [4]

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Amphetamines

Amphetamine Ephedrine Fenfluramine

provide retrospective information about the previous 57years. Axillary and pubic hair can be utilized when the scalp hair is cut short.

Anticonvulsants Antidepressants and antipsychotics Benzodiazepines Cannabinoids Chloroquine Cocaine Doping substances Indinavir Metals Aluminum Arsenic Cadmium Chromium Copper Gold Iron Lead Manganese Mercury Nickel Selenium Silver Thallium Zinc

28.2

History

Hair analysis for the detection of exposure to various substances can be done even years post-mortem. An historic example is the recent discovery that Napoleons death was a result of arsenic poisoning. Measurement of the arsenic content along Napoleons hair shaft permitted his assassination to be traced along a cosmetic and lethal phase. The cosmetic phase consisted of arsenic poisoning over time to weaken Napoleon, making the associated debility appear to be a natural illness. On May 3, 1821, at 5:30 p.m., the lethal phase was carried out: Napoleon was given Calomel (HgCl), a cathartic, and a popular orange-flavored drink called orgeat, which was flavored with the oil of bitter almonds. Together they formed mercury cyanide, which is lethal.

28.3

Pathophysiology of Incorporation of Substances into Hair

Drugs and toxics are incorporated into hair through three different modalities:
Passive diffusion to the hair matrix from the

Nicotine/Cotinine Opiates Codeine Heroin Morphine

Pesticides and persistent organic pollutants Polyamines Putrescine Spermidine Spermine Estrone 17-estradiol DHT Epitestosterone Testosterone Pregnenolone

Sexual hormones

blood and successive incorporation in the hair shaft during keratinization. Within the hair shaft drugs are bound to proteins, melanin or lipids. Correlation between blood and hair concentrations of the substances is not always linear and sometimes the distribution of the substances along the shaft does not correlate well with the time of exposure. Transfer to the formed hair shaft from sebum and sweat. Transfer to the formed shaft from the environment. External contamination may alter hair analysis and therefore decontamination processes are necessary to avoid false-positive results due to passive environmental exposure. Hair decontamination relies on the fact that substances transferred into hair by passive exposure are loosely bound to the surface of the shaft and can be removed by appropriate washing procedures.

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28.4

Techniques of Analysis [8, 9]

28.6

Purposes of Hair Analysis

28.4.1 Hair Sampling

Hair sampling is usually made from the vertex or from the pubis where hair is less contaminated by environmental and cosmetic factors. Hair should be cut close to its point of emergence from the skin to provide information about recent exposure. Hair analysis can be altered by cosmetic procedures such as dyeing, bleaching and permanent waving. About 100200 g of hair is required for analysis. To determine the precise date or duration of intake, one can make use of the knowledge that the mean hair growth rate is 1cm per month, cut the hair sample every 12cm, and analyze the segments separately. Samples can be stored for several months in a dry environment using paper or plastic envelopes.

28.6.1 Forensic Investigations 28.6.1.1 Drug Abuse and Poisoning

Hair analysis permits the tracing of abuse of opiates, cocaine, amphetamines cannabinoids, nicotine, alcohol, and benzodiazepines.

28.6.1.2

Rape Cases

DNA typing can be easily performed on plucked hair and dandruff scales, which can be used to look for the DNA of aggressors in cases where the victims struggled to defend themselves [5].

28.4.2 Decontamination, Extraction, and Detection

Hair washing is important for removing external contaminants that are loosely bound to the hair surface. Washing with organic solvents and phosphate buffers permits the removal of both water-soluble and -insoluble substances. Hair extraction procedures depend on the chemical characteristics of the compound being investigated. These include alkaline digestion, acidic extraction, and enzymatic digestion. Hair analysis methods include immunoassays, gas chromatography, liquid chromatography, mass spectrometry, capillary electrophoresis, and infrared microscopy. Gas chromatography with mass spectrometry offers the best selectivity, sensitivity, and specificity for most substances.

28.6.2 Doping Controls

Although not yet approved by the International Olympic Committee (IOC) hair analysis is very useful for evaluating doping practices in professional athletes. Doping substances that can be detected in the hair include: clenbuterol, corticosteroids, ephedrine, methenolone, nandrolone metabolites, salbutamol, stanozolol, and testosterone. The storage of both nandrolone and its metabolites (norandrosterone and noretiocholanolone) in hair samples permit discrimination of the intake of doping agents from the intake of other 19-norsteroids such as norandrostenedione and norandrostenediol, which are available in over the counter vitamin supplementations [6, 7].

28.6.3 Occupational Medicine

Hair analysis can be used to assess indoor hair pollution by organic chemicals and occupational exposure to toxic substances such as pesticides through inhalation or via the food chain [2]. Substances that can be traced include organochlorine pollutants, organophosphates, carbamates, and pyrethroides. Hair samples can also be utilized to evaluate environmental and occupational exposure of workers to pollutants (lead, nicotine) or metals (nickel, chromium). A strict correlation between mercury level in the hair and fish intake has been recently established.

28.5

Clinical Interpretation

Ruling out external contamination is a main issue. An important criterion for establishing whether active consumption/ingestion has occurred is the presence of both the substance and its metabolites within the sample. Since some drugs such as heroin and cocaine are labile molecules, which can be partially hydrolyzed within the hair fibers, cut-off ratios of metabolite to parent drug have been established (6-monoacetylmorphine:morphine >1.3; benzoylecgonine:cocaine >0.05).

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B. M. Piraccini, M. Pazzaglia, A. Tosti

28.6.4 Prenatal Exposure

R eferences
1. Bernard L, Vuagnat A, Peitavin G et al (2002) Relationship between levels of indinavir in hair and virologic response to highly active antiretroviral therapy. Ann Intern Med 137: 6569 Boumba VA, Ziavrou KS, Vougiolakis T (2006) Hair as a biological indicator of drug use, drug abuse or chronic exposure to environmental toxicants. Int J Toxicol 25: 143163 Caprara DL, Klein J, Koren G (2006) Diagnosis of fetal alcohol spectrum disorder (FASD): fatty acid ethyl esters and neonatal hair analysis. Ann Ist Super Sanita 42: 3945 Daniel CR 3rd, Piraccini BM, Tosti A (2004) The nail and hair in forensic science. J Am Acad Dermatol 50: 25861 Herber B, Herold K (1998) DNA typing of human dandruff. J Forensic Sci 43: 64856 Kintz P, Cirimele V, Jeanneau T, Ludes B (1999) Identification of testosterone and testosterone esters in human hair. J Anal Toxicol 23: 3526 Kintz P, Cirimele V, Ludes B (2000) Discrimination of the nature of doping with 19norsteroids through hair analysis. Clin Chem 46: 20202 Kintz P, Villain M, Cirimele V (2006) Hair analysis for drug detection. Ther Drug Monit 28: 4426 Musshoff F, Madea B (2006) New trends in hair analysis and scientific demands on validation and technical notes. J Forensic Sci Int 5: 24

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Hair analysis can be applied clinically to document in utero exposure to drugs, and other toxic substances including nicotine and alcohol [3].

28.6.5 Patients Compliance to Drug Prescription

Hair analysis is a useful tool to monitor treatment compliance in psychiatric, epileptic, and HIV-positive patients [1]. Another clinical application could be hormone analysis in hair shafts. The dosage of androgens in terminal scalp hair may provide a basis for predicting baldness: the ratio of testosterone: epitestosterone has been demonstrated to be significantly greater in the hair of balding fathers and their sons than in the hair of nonbalding control subjects [6].

2.

3.

4.

5. 6.

28.7

Future Developments

7.

New substances are constantly being measured and hair analysis has the potential for use in other fields of medicine.

8. 9.

Summary for the Clinician

Hair analysis is a necessary complement to blood and urine analysis in order to verify long-term and single exposure to drugs and poisons. Sample collection is noninvasive and easy to perform under conditions that prevent adulteration and substitution.