Automated solid-phase extraction and liquid chromatography-electrospray ionization-mass spectrometry for the determination of flunitrazepam and its metabolites in human urine and plasma samples.
A sensitive and specific method using reversed-phase liquid chromatography coupled with electrospray ionization-mass spectrometry (LC-ESI-MS) has been developed for the quantitative determination of flunitrazepam (F) and its metabolites 7-aminoflunitrazepam (7-AF), N-desmethylflunitrazepam (N-DMF) and 3-hydroxyflunitrazepam (3-OHF) in biological fluids. After the addition of deuterium labelled standards of F,7-AF and N-DMF, the drugs were isolated from urine or plasma by automated solid-phase extraction, then chromatographed in an isocratic elution mode with a salt-free eluent. The quantification was performed using selected ion monitoring of protonated molecular ions (M+H(+)). Experiments were carried out to improve the extraction recovery (81-100%) and the sensitivity (limit of detection 0.025 ng/ml for F and 7-AF, 0.040 ng/ml for N-DMF and 0.200 ng/ml for 3-OHF). The method was applied to the determination of F and metabolites in drug addicts including withdrawal urine samples and in one date-rape plasma and urine sample.
Jourdil
N
N
Laboratory of Pharmacology, Grenoble University Hospital, BP 217, 38043 Grenoble Cedex 9, France.
Bessard
J
J
Vincent
F
F
Eysseric
H
H
Bessard
G
G
D000328:Adult; D014151:Anti-Anxiety Agents / Q000097:blood / Q000378:metabolism* / Q000652:urine; D001331:Automation; D002138:Calibration; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D005445:Flunitrazepam / Q000097:blood / Q000378:metabolism* / Q000652:urine; D006801:Humans; D008297:Male; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D019966:Substance-Related Disorders / Q000652:urine
D014151:Anti-Anxiety Agents; D005445:Flunitrazepam