Page 157 - Advances in Forensic Applications of Mass Spectrometry - Jehuda Yinon

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Figure 3.5 shows a chromatogram representative of an authentic hair
specimen with the corresponding concentrations: 2.5, 3.2, 10, and 0.6 pg/mg
for testosterone, epitestosterone, DHEA, and DHT, respectively.

3.3.2 Detection of Corticosteroids

Cortisone and hydrocortisone, naturally occuring hormones, influence
metabolism, inflammation, and electrolyte and water balance. Their synthetic
derivatives are used in therapeutic medicine for their antiinflammatory and
immunosuppressive actions. They are used in certain sports to improve the
performances of the athletes (euphoria, motor activity).
Cirimele et al. published in 1999 the first identification of such a drug,
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in this case, prednisone, in the hair of a subject treated for years. A 50-mg
hair specimen was incubated overnight in Sorensen buffer, then extracted by
solid-phase extraction using an Isolute C18 column. Prednisone was detected
by LC/MS at 1280 pg/mg. Using the same preparation technique, and cortisol-
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d as an internal standard, the same group published several months later
3
a screening procedure for 10 corticosteroids, with detection limits in the range
of 30 to 170 pg/mg. Two applications were documented for prednisone and
beclomethasone, identified in hair at 140 and 230 pg/mg, respectively.
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Using a 2 mm i.d. column, Cirimele et al. demonstrated in 10 patients
treated with prednisone a low but not insignifiant correlation (R = 0.578,
2
p < 0.03) between the total amount of ingested drug and the measured con-
centrations in hair. The procedure was sensitive enough to detect prednisone
in the hair of patients treated with a low 5 mg/d dose.
Repetitive abuse of corticosteroids by athletes can be demonstrated by
segmental analysis along the hair shaft, in contrast to ponctual urinalysis. A
single treatment of about 1 week will make positive a single 1-cm segment,
while long-term abuse will lead to the identification of the corticoid(s) in
several segments. For such an application, particularly in the case of longi-
tudinal surveys of athletes, hair analysis appears as the solution of choice to
document doping practices. It has been demonstrated that a single oral
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therapeutic treatment with 4-mg/d betamethasone for 9 consecutive days is
detectable through hair analysis. The drug tested at a concentration of 4.7
pg/mg in the corresponding hair segment, whereas no betamethasone could
be identified in the distal hair strand. Extraction of the drug was classic from
this group, however, to enhance sensitivity a MIC 15 CP Nucleosil C18, 150
¥ 1 mm i.d. column was used.
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A confirmatory method was developed for the quantitative determina-
tion in hair of the most common corticosteroids used as doping agents by
athletes. Drugs were extracted from 50 mg of powdered hair by methanolic
extraction followed by a solid-phase extraction on a C18 column. Detection
was performed with an electrospray ionization mass spectrometer in the

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