1 Institute of Legal Medicine, Humboldt-University, Hannoversche Straße 6, 10115 Berlin, Germany
Received 5 August 2002; in revised form 11 October 2002; accepted 28 October 2002
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ABSTRACT |
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INTRODUCTION |
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The analysis of human hair for illicit and therapeutic drugs is now a routine procedure in many laboratories. Comprehensive reviews have been published about the anatomical, physiological and biochemical basis of hair analysis as well as the analytical methods used and the application of this methodology (Kintz, 1996; Pragst et al., 1998
; Sachs and Kintz, 1998
). Usually scalp hair is investigated. The use of hair from additional or alternative sites may be helpful if: (1) the head has been shaved (no scalp hair available); (2) the scalp hair is too short or it has been treated frequently by invasive cosmetics (dyeing, bleaching, permanent wave); (3) an additional sample is needed to confirm the scalp hair results; or (4) there is a suspicion of external contamination with the substances to be analysed (Mangin, 1996
). In most such cases, pubic hair was analysed, but beard, axillary or body hair has been used only on occasions. However, the analyte concentrations in these alternative samples may be different from those in scalp hair, since there are substantial inter-site differences in hair growth rate and growth cycle, in the nature and activity of sweat and sebum glands, and in hair thickness (Ebling, 1987
). Furthermore, there are differences with respect to atmospheric exposure and other external influences, as well as to hair care/use of cosmetics.
Drug concentrations in scalp, pubic and axillary hair have been compared by several authors (Mangin and Kintz, 1993; Offidani et al., 1993
; Kintz and Mangin, 1994
; Mangin, 1996
; Kosuge et al., 1998
). As a rule, the concentrations of opiates and cocaine decreased in the order pubic > scalp > axillary hair. Although beard hair was analysed several times for drugs in the context of the time of appearance of the analyte after drug intake (e.g. Cone, 1990
), no comparison of analyte concentrations in this site with hair from other sites has been described.
In order to investigate the extent to which hair samples from sites other than scalp hair could be analysed in cases of suspected alcohol misuse, FAEE concentrations in scalp and pubic hair from 22 deaths, of which 18 had an alcohol anamnesis, and of one teetotaller and five moderate social drinkers were measured. Axillary hair, beard hair and different body hair was also analysed in some cases.
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MATERIALS AND METHODS |
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The protocol was approved by the ethics commission of the University Hospital Charité of Humboldt University.
Sample preparation and measurement of FAEE by gas chromatographymass spectrometry (GCMS) and head-space solid-phase microextraction (HS-SPME)
The analytical procedure and the mass spectra of the four FAEE (the myristate, palmitate, oleate and stearate ethyl esters) and of the corresponding deuterated internal standards (d5-FAEE) have been described in detail (Pragst et al., 2001). Briefly, 2050 mg of the washed hair sample were cut to pieces of about 1 mm length and weighed. Then, 0.5 ml of dimethylsulphoxide, 2 ml of heptane and 40 ng of each of the four d5-FAEE were added in 20 µl of chloroform. The mixture was shaken (15 h, 25°C), centrifuged, the heptane phase separated, the solvent evaporated to dryness in a nitrogen stream, and the residue analysed by HS-SPME and GC-MS-SIM (selected ion monitoring). The limits of detection and limits of quantification were: ethyl myristate, 0.015 and 0.05 ng/mg; ethyl palmitate, 0.02 and 0.07 ng/mg; ethyl oleate, 0.04 and 0.12 ng/mg; and ethyl stearate, 0.01 and 0.04 ng/mg, respectively. Assay reproducibility was assessed by replicate analysis (n = 10) of the one hair sample. The standard deviations were: ethyl myristate (0.067 ng/mg), 15.7%; ethyl palmitate (0.796 ng/mg), 6.6%; ethyl oleate (0.387 ng/mg), 5.9%; and ethyl stearate (0.133 ng/mg), 3.5%.
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RESULTS AND DISCUSSION |
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From the study data in Table 1, it is clear that, in most cases, the FAEE concentrations in scalp hair differed considerably from the concentrations in hair from other sites in the same subject, but, nevertheless, excessive alcohol consumption was clearly indicatedthis is shown for a moderate social drinker and two alcoholics in Fig. 1
. Some features of hair growth and hair properties, which could contribute to the explanation of these differences in FAEE concentrations between samples from different sites, are given in Table 2
. Human hair follicles and their associated sebaceous and apocrine glands are generally regulated by androgens and do not behave identically, but vary greatly between body site (Takayasu et al., 1980
; Ebling, 1986
). Different growth rate and growth cycle, as well as different size and activity of the glands, could explain the differences in FAEE concentrations observed.
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Beard hair
All 16 beard hair samples were collected from the chin. Of these, seven showed lower FAEE concentrations (beard/scalp FAEE concentration ratio, 0.450.89) and nine higher FAEE concentrations (beard/scalp, 1.25.7), compared with scalp hair. With alcoholics particularly (generally much higher FAEE concentrations in beard hair, e.g. S247/01), it is possible that the beard is frequently wetted by alcoholic beverages from which ethanol could be absorbed topically and transformed into FAEE in the sebum glands. A similar phenomenon may occur after use of hair lotions containing high ethanol concentrations (Hartwig et al., 2003). Assuming again a cut-off of 1.0 ng/mg, the interpretation of the beard hair concentrations with respect to excessive alcohol consumption leads to the same 11 positive results as with scalp hair.
Body hair
In eight cases, hair samples from the chest, arms or legs, and in one case also from the eyebrow, were analysed. Of all kinds of human hair, body hair varies most in growing conditions, length and diameter (Ebling, 1990). The FAEE concentrations indicated excessive alcohol consumption in four cases, in the same way as the samples from the other sites by CFAEE > 1.0 ng/mg. However, the number of samples was too small for firm conclusions to be drawn.
In conclusion, FAEE contain the ethyl group of ethanol and, therefore, are direct markers of ethanol exposure. They are deposited in all kinds of human hair after alcohol consumption. Chronic excessive alcohol consumption can be detected reliably by the presence of high FAEE concentrations in scalp, pubic, axillary, beard as well as body hair, although the actual values obtained vary markedly between different sites in the same subject. The analysis of one or more hair samples from different body sites serves to confirm scalp hair results and helps to avoid errors in interpretation caused by the use of hair cosmetics. We believe that the results of this preliminary study warrant further research on larger samples of both alcoholic and control subjects.
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ACKNOWLEDGEMENTS |
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FOOTNOTES |
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REFERENCES |
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