ISSN: 0022-1198
CODEN: JFSCA
Page Count: 9

Analysis of Biofluids for Flunitrazepam and Metabolites by Electrospray Liquid Chromatography/Mass Spectrometry
Wagner, JR
College of Medicine, University of California-Irvine, CA

LeBeau, MA
Chemistry Unit, FBI Laboratory, Federal Bureau of Investigation, DC

Montgomery, MA
Chemistry Unit, FBI Laboratory, Federal Bureau of Investigation, DC

Miller, ML
Chemistry Unit, FBI Laboratory, Federal Bureau of Investigation, DC

(Received 25 June 1999; accepted 5 November 1999)

Abstract

A rapid and sensitive liquid chromatography/electrospray ionization mass spectrometry (LC/MS) procedure has been developed for the analysis of biofluids containing flunitrazepam and its metabolites. Specimens were spiked with deuterated analogs of the analytes. Urine specimens were enzymatically hydrolyzed and blood specimens were untreated. Extractions were carried out using CleanScreen DAU SPE cartridges. The drugs were separated on a C18 column using a methanol-water-ammonium hydroxide (60:40:0.03 v/v) mobile phase. After determination of base peaks using full scan mass spectrometry, the mass spectrometry method was optimized to operate in selected-ion monitoring (SIM) mode for the base peak of each analyte. Positive findings were confirmed by LC/MS/MS using the same mobile phase and column.

This analytical procedure allows for the detection of low levels of flunitrazepam and metabolites in biofluids. It is useful for ascertaining the role of flunitrazepam in cases of drug-facilitated sexual assault.

Keywords:
forensic science, flunitrazepam, Rohypnol®, drug-facilitated, sexual assault, drug rape, date rape, forensic toxicology

Paper ID: JFS14840J
DOI: 10.1520/JFS14840J
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