SOFT 2004 Meeting Abstracts - Society of Forensic Toxicologists

M30
DOPING STEROID ANALYSIS IN NAIL CLIPPINGS
Kenichi Takaichi l ., R.A. Anderson I and D. Thieme 2 : IDepartment of Forensic Medicine and Science, University of
Glasgow, Glasgow G 12 8QQ, United Kingdom and 21nstitute of Doping Analysis/Sports Biochemistry, Dresdner Strasse
12, D-01731 Kreischa, Germany.
Extraction ofanalytes from fingernail is difficult due to its hard keratin composition. Our group has shown the feasibility
of drug analysis in nail using a cryogenic grinding technique, including cannabinoids and opioids (I). In particular,
cryogenic grinding has been shown to allow the extraction and analysis of intact esters such as diamorphine by avoiding
the alternative alkaline hydrolysis step. To date, few papers have reported the detection ofsteroids in nail (2). By contrast,
several papers have reported the detection and analysis of anabolic steroids in hair.
In the present study, cryogenic grinding was used to prepare nail clippings from doping abusers for extraction of
endogenous and anabolic steroids to show that both can be detected, as in hair. This method was compared to blood
(plasma) and urine samples that had been analysed at the same time.
Fingernail clippings from users of anabolic steroids (including testosterone esters, stanozolol and methenolone acetate)
were obtained from the Institute of Doping Analysis/Sports Biochemistry, Kreischa. Blank nail samples were obtained
from volunteers. The nail samples were first decontaminated by washing with 0.1% sodium dodecyl sulfate, water (x 3)
and methanol (x 3). After drying, the samples were pulverised for 4 min in a liquid-nitrogen cryogenic miII (SPEX
CertiPrep 6750 Freezer Mill). The powdered nail was then extracted with methanOl/ethyl acetate (7:3 v/v, 7 mL).
Deuterated anabolic steroids (5a-estran-3B-oI-17-one-d 3 , testosterone-d 3 , and stanozolol-d 3 ) and medroxyprogesterone
were used as internal standards.
The extracts were converted to TMSi derivatives with MSTFAlNH 4 112-mercaptoethanol and analyzed by GC-MS in the
EI + full scan and SIM modes, on a Finnigan Trace instrument equipped with an HP-5 column (30 m x 0.32 mm i.d.; film
thickness 0.25 J.lm) with temperature programming from 180°C to 240 °C at 3 °C/min, 240°C to 300 °C at 5 °C/min, and
final temperature held for 10 min.
In this initial study endogenous steroids (androsterone (And), etiocholanolone (Etio), dehydroepiandrosterone (DHEA),
epiandrosterone (epi-And), epitestosterone (epi-Test) and testosterone (Test» were identified and quantified in the nail
samples from both steroid users and non-users.
No. And Etio DHEA epi-And epi-Test Test
Suspect # I (Nail) 4.1 0.0 5.6 3.3 3.0 4.3
Suspect #2(Nail) 4.7 0.0 3.0 1.9 1.5 3.9
Suspect #3(Nail) 9.6 0.0 25.3 14.3 11.0 10.6
Suspect #4(Nail) 12.8 0.0 12.0 6.2 10.9 25.0
Suspect #5(Nail) 1.7 0.0 1.8 0.8 0.6 0.6
Healthy # 1 (Nail) 1.5 0.0 1.4 0.7 0.6 6.5
Healthy #2(Nail) 8.4 0.0 4.5 1.7 2.6 l.l
Healthy #1(Plasma) 30322.9 54222.5 11218.3 Trace 222.2 3583.8
Healthy #2(Plasma) 9923.3 2966.7 100.6 Trace 200.2 196.0
Healthy # I (Urine) 351.6 392.5 11.2 Trace 9.0 11.0
Healthy #2(Urine) 926.8 868.7 115.8 Trace 7.4 13.6
* Suspect #3
Test 292 pg/mL in urine,
Test-enanthate, decanoate
in hair were 20 and 18
pg/rng, respectively.
* Suspect #4
Test 30 pglrnL in urine,
Test-enanthate,
phenyl-propionate,
decanoate, and isocaproate
in hair were 25, 425, 220,
and 340 pg/rng,
respectively.
In general, endogenous steroid concentrations in nail were low, in the pgjmg range. Also, elevated concentrations of
testosterone in nail were positively associated with high concentrations in plasma and urine. However, although the
analytical results provided evidence for the presence of anabolic steroids in the samples from steroid users, including
testosterone and testosterone esters at low concentrations, it has not yet been possible to confirm this due to interference
from other endogenous substances. Nail remains a potential, but still to be confirmed, alternative biological specimen to
hair for the detection ofpast exposure to doping steroids.
References:
(1) K. Takaichi, N.P. Lemos, R.A. Anderson, Analysis of Opiates in Nail Clippings from Chronic Heroin Abusers,
presented at the 39 th Annual TIAFT Meeting, Prague, 200 I.
(2) M.H. Choi, Y.S. Yoo, B.C. Chung, Measurement of testosterone and pregnenolone in nails using GC-MS.,
J. Chromatogr., B 754, 495·501 (2001).
Keywords: Nail Analysis, Doping, Anabolic Steroids
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