Free Content Serum and Urine Concentrations of Flunitrazepam and Metabolites, after a Single Oral Dose, by Immunoassay and GC–MS

Authors: Snyder H.1; Schwenzer K.S.1; Pearlman R.1; McNally A.J.1; Tsilimidos M.1; Salamone S.J.1; Brenneisen R.2; ElSohly M.A.3; Feng S.3

Source: Journal of Analytical Toxicology, Volume 25, Number 8, November/December 2001 , pp. 699-704(6)

Publisher: Preston Publications

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Abstract:

A clinical study was conducted to assess the ability of commercially available immunoassays to detect flunitrazepam (FNP) in plasma and urine samples and to compare the results with those obtained by gas chromatography–mass spectrometry (GC–MS). The clinical study consisted of four individuals (two male and two female) who had taken a single 2-mg dose of FNP. Serum was collected over a 48-h period and urine was collected over a 72-h period. The serum and urine samples were analyzed by the COBAS® INTEGRA Serum Benzodiazepines assay (SBENZ), the TDx serum and urine Benzodiazepines assay, and GC–MS. The GC–MS procedure was developed for analysis of FNP and metabolites in plasma and urine using an acid hydrolysis step resulting in the formation of specific benzophenones corresponding to FNP and its metabolites. The relative sensitivities of the assays for the detection of FNP and metabolites in serum and urine were GC–MS > SBENZ > TDx. The immunoassay results for serum samples showed peak concentrations of FNP metabolites at 8 h after FNP ingestion for three individuals and at about 1 h for the fourth individual. The GC–MS, SBENZ, and TDx urine immunoassays detected drug above the stated limit of detection (LOD) in 44, 41, and 35 serial FNP urine samples, respectively. FNP metabolites were detected in urine samples with all three assays for up to 72 h after a 2-mg dose. The improved detection rate with the SBENZ assay as compared to the TDx assay is likely explained by its higher cross-reactivity with the major metabolite, 7-amino-flunitrazepam (7-amino-FNP), and its lower LOD.

Language: English

Document Type:

Affiliations: 1: Roche Diagnostic Systems, Somerville, New Jersey 08876 2: University of Bern, Bern, Switzerland 3: ElSohly Laboratories, Incorporated, Oxford, Mississippi 38655

Publication date: 2001-11-01

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  • The Journal of Analytical Toxicology (JAT), established in 1977 and published 9 times a year, is the international source covering a broad range of clinical, forensic, and industrial laboratory topics regarding the isolation, identification, and quantitation of potentially toxic substances.

    With an emphasis on practical application, JAT articles provide improved and novel techniques for use in clinical, forensic, workplace, sports testing (doping), and other toxicology laboratories. Articles describe newly developed methods in immunoassay testing, gas chromatography, liquid chromatography, mass spectrometry, atomic absorption spectrometry, solid and liquid phase extraction techniques, and other analytical approaches. Worldwide readership includes toxicologists, pathologists, chemists, clinicians, researchers, and educators working in medical examiner and law enforcement laboratories, hospitals, university and independent analytical laboratories, as well as the drug manufacturing industry.

    Each year in October, we publish a special issue from the Society of Forensic Toxicologists.

    JAT, as determined by ISI Citation Index, is one of the two most referenced international journals in forensic science.

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