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Published:
Journal of Analytical Toxicology,
ISSN 0146-4760,
Volume 26, Number 4, May/June
pp. 211-215
Detection of Flunitrazepam and 7-Aminoflunitrazepam in
Oral Fluid after Controlled Administration of Rohypnol®
Nele Samyn[1], Gert De Boeck[]1, Vincent Cirimele[2],
Alain Verstraete[3], and Pascal Kintz[2]
[1]National Institute of Criminalistics and Criminology, Toxicology Section,
Brussels, Belgium;
[2]Institut de Médecine Légale, Strasbourg, France; and
[3]Laboratory of Clinical Biology-Toxicology, Ghent University Hospital, Ghent,
Belgium
Although administered as a
short-acting hypnotic for sleeping disorders, flunitrazepam, often in combination
with alcohol or other drugs, was one of the most frequently abused benzodiazepines
over the last 10 years. It has been reported in cases of driving under the influence,
and its use is associated with marked psychomotor impairment. Studies over the
last five years have investigated the use of oral fluid as an alternative matrix
to blood and urine, especially when non-intrusive and quick sampling procedures
are important (e.g., screening for drugs of abuse at the roadside and screening
and confirmatory workplace drug testing). In this study, Rohypnol (flunitrazepam)
was administered to four healthy volunteers, and oral fluid samples were collected
by spitting into a polypropylene tube at fixed times between 0 and 6 h after
the intake of a tablet of 1 mg. A specific and very sensitive method was developed,
both for flunitrazepam and for its main metabolite 7-aminoflunitrazepam, based
on solid-phase extraction of the oral fluid samples, stored at +4°C, and
gas chromatographic–mass spectrometric analyses using negative chemical
ionization with methane as the ionization gas. The heptadeuterated parent compound
and metabolite were used as internal standards. The respective limits of detection
and quantitation were 0.05 µg/L and 0.1 µg/L for flunitrazepam,
and 0.1 and 0.15 µg/L for 7-aminoflunitrazepam. The parent drug could
only be detected when the analyses were performed within 12–24 h after
collection of the oral fluid samples or when 2% of NaF was added to the collection
tubes. The stability of flunitrazepam in oral fluid was poor, even at +4°C,
when no NaF was added to the sample. In any case, concentrations remained below
1 µg/L. The metabolite was detected in slightly higher concentrations,
with or without the presence of NaF, reaching a maximum of 1–3 µg/L
within 2–4 h after administration. In all cases the drug was detectable,
but at extremely low concentrations, for 6 h after intake of a normal dose of
Rohypnol and it will be an analytical challenge to come up with a sufficiently
sensitive onsite test for low-dose benzodiazepines in oral fluid. Reproduction
of editorial content of this journal is prohibited without publisher’s
permission.
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