SOFT 2004 Meeting Abstracts - Society of Forensic Toxicologists

M9
SCREENING OF BENZODIAZEPINES AND HYPNOTICS IN HAIR BY LC-MSIMS:
APPLICATION TO FOUR DRUG-FACILITATED CRIMES
M. VilIain 1 *, M. Cheze 2 , B. Ludes 1 , P. Kintz!; 1 Institut de Medecine Legale, II rue Humann, 67000
Strasbourg, France and 2 ToxLab, 7 rue Jacques Cartier, 75018 Paris, France
The use of a drug to modifY a person's behavior is not a recent phenomenon, but reports dealing with drugfacilitated
crimes (sexual assaults, robbery ... ) are constantly increasing. Among drugs involved in such
cases, benzodiazepines (flunitrazepam, bromazepam, ... ) and hypnotics (zopiclone, zolpidem) are largely
concerned.
In most cases and due to a long delay between the event and the claim to the police, blood and even urine
samples are of little value. This is the reason why this laboratory developed a original approach based on
hair testing. In practice, hairs of the victim are collected about one month after the alleged offense,
segmented and analyzed by LC-MSIMS. In positive cases, the drug is supposed to be found only in the
segment corresponding to the period ofthe event.
The Hair strand is twice decontaminated using methylene chloride and then segmented (3 segments of 2
cm). Each segment is cut into small pieces «lmm). About 20 mg are overnight incubated in phosphate
buffer at pH 8.4, in the presence of 1 ng of diazepam-ds used as internal standard (IS). After a liquid-liquid
extraction with a mixture of methylene chlorideldiethyl ether (80/20) and evaporation to dryness, the
residue is reconstituted in 50 J.1L of acetonitrile/water (50/50).
Chromatography is achieved using a XTerra MS CI8 column (100 x 2.1 mm, 3.5J.1m) eluted with a
gradient of acetonitrile and formate buffer delivered at a flow rate of 0.2 mLlmin.
A Quattro Micro triple-quadrupole mass spectrometer is used for analyses. Ionization is achieved using
electrospray in the positive ionization mode (ES+). For each compound, detection is related to two
daughter ions.
To demonstrate the applicability ofthis method, 4 expertises are describe as follows:
1. A-21 year old girl was offered a coffee that made her unconscious. The victim's hair, collected 15
days after the alleged offense, was tested by LC-MSIMS and the first 2 cm segment was positive
at a concentration of4.4 pg/mg whereas the 2 other segments were zolpidem free.
2. A 16-year old girl claimed to have been raped during an afternoon, while sedated. Hair was
sampled 9 weeks after the alleged offense and the first 3 cm segment was positive for zopiclone at
a concentration of 4.2 pg/mg, the second (3 to 5 cm) at 1.0 pg/mg, whereas the last segment (5 to
7 cm) was zopiclone free.
3. A 39-year old woman felt sleepy for 24 hours after having drunk a coffee. Hair was collected I
month after the event and the proximal 2 cm-Iong segment was positive for bromazepam at 10.3
pg/mg and the other segments (2-4 and 4-6 cm) remained negative.
4. A woman was confined illegally for 12 days, repetitively raped and forced to sign .cheques, while
being under the influence of an hypnotic. Hair was positive for 7-aminoflunitrazepam in the
proximal segment (0 to 3 cm) at 31.7 pg/mg, in the second segment (3 to 6 cm) at 2.0 pg/mg and
was negative in the distal one (6 to 9 cm).
In drug-facilitated crimes, hair is a very useful alternative specimen, and the concentrations measured, in
the pg/mg range, require tandem mass spectrometry.
Keywords: Hair, LC-MS/MS, drug-facilitated crime
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