SOFT 2004 Meeting Abstracts - Society of Forensic Toxicologists

M20
DRUG-FACILITATED CRIMES - INTEREST OF TANDEM MASS SPECTROMETRY AND
HAIR ANALYSIS - STATISTICS OF DRUGS IMPLICATED DURING A YEAR OF WORK AT
TOXLAB
Duffort G.*, Cheze M., Pepin G.
TOXLAB Laboratory, 7 rue Jacques Cartier -75018 Paris - FRANCE
AIMS: Drug-facilitated crimes cases are often difficult to resolve, from an analytical point of view, since
the drugs administered, largely benzodiazepines or hypnotics, are generally given as one therapeutic dose,
and most often have a short half-life elimination. In France, for these kind of aggression cases, the police
directs the victim towards a hospital or, in big cities, to an emergency forensic service office. Ifnecessary,
after the clinical exam, blood and sometimes urine samples are taken. Using HPLCIDAD and/or GCIMS,
immunochemical as well as toxicological screenings are performed on these samples. The immunochemical
techniques constantly produce negative results. HPLCIDAD and GCIMS do not permit most basic
molecules used in drug-facilitated crimes to appear because the aggression reports, and then the samples,
are often late and thus substances are undetectable with those techniques. This is why we have developed
an HPLC-MSIMS method for the benzodiazepines and hypnotics determination in biological fluids and
hair. By that way, we can reach the detection limits of the nglmL order in biological fluids and of the
pglmg order in hair. Human hair is sampled by us, about one month after the exposure. The sequential hair
analysis by LC-MSIMS allows to affirm that the identified molecules in blood and/or in urine were in fact
from an isolated single dose and not from a medical treatment, so it brings evidence for drug-facilitated
crimes.
METHODS: The detection of main benzodiazepines in biological fluids were performed by LC-MS/MS
after an extraction of I mL with Toxitube A ® (Varian). Hair was segmented, decontaminated, cut, then 20
mg were extracted with deuterated standard, whether one night incubation in ImL Soerensen buffer (PH
7.6), or whether, for aminometabolites, at 95° for 15 minutes in NaOH O,IN. A liquid-liquid extraction was
performed with dichloromethane/ether (80/20). The organic phase was filtered before being evaporated.
The dry extract was reconstituted by 60JlL of ACNlMeOH (50/50) and injected into the LC-MS/MS
(ThermoElectron). The liquid chromatography was carried out on an Uptisphere ODB C18 51-! 150 x 2mm
column (Interchim) with a gradient of acetonitrile and formate 2mM pH3 buffer. The run was 15 minutes
long. The mass tandem TSQ Quantum detection allowed the simultaneous determination of 27
benzodiazepines in SRM mode with two runs.
RESULTS: The detection limits reached by these techniques, are in the order or inferior to 0.5 nglmL in
urine and about 1 pglmg in hair for the 27 benzodiazepines studied. We present two cases of drug-facilitated
crimes in which cIonazepam and flunitrazepam were found in blood and urine samples. In these two cases,
a single dose of these molecules appeared on the specific segment of hair corresponding to the period of the
offense. From June 2003 to May 2004 we treated 90 cases of presumed drug-facilitated crimes, for which
we had urine and/or hair samples. In 35.5% ofthese cases, the benzodiazepines or hypnotics were detected.
Among these, a single dose, taken without someone's knowledge, appeared in 65.5%. In the 90 cases,
35.5% of the victims were vulnerable following an alcohol and/or illegal drugs consumption; in 22.2% the
investigation showed that it was not a drug-facilitated crimes and 6.7% were not able to be solved.
CONCLUSIONS: This technique produces an analytical response that is essential to police and justice
authorities. It brings out criminal acts and produces the scientific proof of drug-facilitated crimes with
benzodiazepines and hypnotics as shown by our statistics and the two specific cases cited.
Keywords: Benzodiazepines, LC/MS/MS, Hair Analysis
Page 303