Page 150 - Advances in Forensic Applications of Mass Spectrometry - Jehuda Yinon

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female subjects. After decontamination with dichloromethane, 100 mg of hair
was incubated in 1 M NaOH in the presence of testosterone-d . After neu-
3
tralization, the extract was purified using solid-phase extraction with Isolute
C18 columns followed by liquid–liquid extraction with pentane. After silyla-
tion, the drugs were analyzed by GC/MS. Concentrations for DHEA were in
the range 1 to 7 pg/mg (mean 4 pg/mg) and 0.5 to 11 pg/mg (mean 5 pg/mg)
for the males (n = 15) and females (n = 12), respectively. Concentrations for
testosterone were in the range 0.5 to 12 pg/mg (mean 4 pg/mg) and not
detected to 2 pg/mg for the males (n = 41) and females (n = 12), respectively.
Unlike testosterone in urine, the interpretation of concentration findings
in hair can be difficult and critical. The range between physiological concen-
trations of testosterone and those found in abusers seems to be rather small.
Therefore, in complement of testosterone determination, the identification
of unique testosterone esters in hair enables an unambiguous charge for
doping because the esters are certainly exogenous substances. This was largely
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10
developed by Thieme et al. and Gaillard et al. Recently Rivier claimed that
8
although hair analysis alone cannot be useful for screening purposes, it could
become in the future a possibly useful technique for obtaining additional
information on long-term testosterone abuse.
8
Thieme et al. published in 2000 a complete analytical strategy for detect-
ing anabolics in hair. The preparation of the sample was carried out by a
methanol extraction step with sonication for all the anabolics except for
stanozolol which was incubated in NaOH. Extensive cleanup procedures were
employed such as HPLC and solid-phase extraction, followed by derivatiza-
tion to form the enol-TMS derivatives. Drugs were identified either by
GC/MS/MS or GC/HRMS. Metandienone and its metabolite, 6b-hydroxy-
metandienone, stanozolol and its metabolite 3¢-hydroxy-stanozolol, mester-
olone, metenolone enantate, and nandrolone decanoate, and several
testosterone esters such as propionate, isocaproate, decanoate, and phenyl-
propionate were identified in the hair of several bodybuilders.
Gaillard et al. developed a method for testing both the anabolic steroids
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and their esters. A 100-mg amount of powdered hair was first treated with
methanol for extraction of esters, then alkaline digested with 1 M NaOH for
the recovery of the other drugs. These preparations were extracted with ethyl
acetate, pooled, then finally highly purified using a twin solid-phase extrac-
tion on amino and silica cartridges. After silylation, drugs were detected by
GC/MS/MS. Figure 3.1 shows the MS/MS chromatograms of an extract of a
human hair spiked with 50 pg/mg of each one of a series of drugs. Nandrolone
and testosterone undecanoate were identified in the hair of 2 athletes at 5.1
and 15.2 pg/mg, respectively.
A sensitive, specific, and reproducible method for the quantitative deter-
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mination of stanozolol in human hair was developed by Cirimele et al. The

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