Page 159 - Advances in Forensic Applications of Mass Spectrometry - Jehuda Yinon

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negative ion mode. The limits of sensitivity were about 100 pg/mg. Hair from
athletes revealed the presence of hydrocortisone acetate, methylprednisolone,
triamcinolone acetonide, and dexa/betametasone at 430, 1350, 280, and 1310
pg/mg, respectively. According to the authors, who tested in parallel the
corresponding urine specimens, the comparison of the results demonstrated
once again the dramatic complementary use of urinalysis and hair analysis. 23

3.3.3 Detection of bb bb-Adrenergic Stimulants
Because of their sympatomimetic properties (stimulant effects) and their
activity as anabolic agents at higher dosages, b2-agonists are banned. How-
ever, salbutamol is permitted by inhalers only and must be declared in
writing prior to the competition. To date, only three studies have been
published in the literature for these drugs in human hair, two for clenbuterol
24
and one for salbutamol, respectively. In their paper, Gleixner et al. identified
clenbuterol after incubation in 1,4-dithiothreitol, NaOH, and tertiary butyl-
methyl ether by enzyme immunoassay (EIA), with confirmation by
HPLC/EIA. Clenbuterol accumulated in hair after 10 mg/d for 25 d at con-
centrations ranging from 23 to 161 pg/mg, with relatively high concentra-
tions in dark hair. The drug was also found in the hair from two bodybuilders
at 50 and 92 pg/mg, respectively.
25
Machnik et al. tested clenbuterol in the hair of four females who had
therapeutically taken the drug as a tocolyticum. Hair was incubated in 1 M
KOH, and the drug extracted with tertiary butylmethyl ether, followed in some
cases by immunoaffinity chromatography, then derivatizated with MSTFA-
ammonium iodide-TMS ethanethiol. High-resolution mass spectrometry was
used to identify clenbuterol. Limit of detection was about 0.8 pg/mg. The
levels of clenbuterol determined in hair ranged from 2 to 236 pg/mg.
26
A screening procedure was developed for simultaneous identifications
of b2-agonists and b-blockers. The procedure involved overnight incubation
in 0.1 M HCl, followed by neutralization, solid-phase extraction with an
Isolute C18 column, derivatization with trimethylboroxine/ethyl acetate, and
GC/MS detection. Limits of detection were 2 pg/mg for both salbutamol and
clenbuterol. In nine asthmatic patients, the salbutamol concentrations in hair
were in the range 27 to 192 pg/mg. In two asthma deaths, salbutamol con-
centrations in hair were 210 and 87 pg/mg, respectively. Finally, the labora-
tory identified salbutamol in the hair of a swimmer, which was positive in
urine, at a level of 71 pg/mg. Figure 3.6 shows the mass chromatogram and
the electron ionization mass spectrum of the extracted hair specimen.
Most of the positive specimens are reported for salbutamol (46% of the
total urines in 1999 for the IOC laboratory in France). However, as this drug
is permitted for specific therapeutic purposes, together with a medical pre-
scription, it appears very easy to evade the test, and almost all cases are

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