Urinary screening for adinazolam and its major metabolites by the Emit d.a.u. and FPIA benzodiazepine assays with confirmation by HPLC.
Adinazolam is a triazolobenzodiazepine, currently under clinical investigation, that possesses antidepressant and anxiolytic activity. It has a short half-life (less than 3 h), and less than 2% of an oral dose is excreted unchanged. The major urinary metabolite is N-desmethyladinazolam, and minor metabolites are estazolam and alpha-OH-alprazolam. The objective of this study was to characterize the reactivity of adinazolam, N-desmethyladinazolam, and estazolam in the Emit d.a.u. benzodiazepine assay and the Abbott TDx urine (FPIA) benzodiazepine assay. N-desmethyladinazolam and estazolam gave an equivalent response to the Emit cutoff calibrator (300 ng/mL) at 100-200 ng/mL, and adinazolam gave an equivalent response at 200 ng/mL. By FPIA, N-desmethyladinazolam and adinazolam had equivalent net polarization values as the 300-ng/mL low control at 500-1000 ng/mL, and estazolam gave a positive response at 300 ng/mL. Six volunteers received single oral doses of 10, 30, and 50 mg of adinazolam. Urine specimens (N = 7) were collected from 0 to 36 h post-administration. By Emit, all urine specimens at all doses were positive from 2 to 36 h, and all FPIA analyzed specimens were positive from 2 to 24 h. Confirmation testing was performed by HPLC by analyzing for N-desmethyladinazolam. All urine specimens were confirmed positive for N-desmethyladinazolam (greater than 200 ng/mL) except for the blank specimens (time = 0) and 7 of 18 specimens collected 36 h post-administration. In conclusion, both immunoassay screening assays are acceptable for detecting the presence of adinazolam in human urine for up to 24 h after a single oral dose of 10-50 mg.
Fraser
A D
AD
Toxicology Laboratory, Victoria General Hospital, Halifax, Nova Scotia, Canada.
Isner
A F
AF
Bryan
W
W
D000525:Alprazolam / Q000031:analogs & derivatives / Q000652:urine; D014151:Anti-Anxiety Agents; D001569:Benzodiazepines / Q000652:urine*; D002851:Chromatography, High Pressure Liquid; D004187:Disaccharides; D004949:Estazolam / Q000652:urine; D005455:Fluorescent Antibody Technique; D005965:Glucuronates; D006801:Humans; D007118:Immunoassay / Q000379:methods*; D007124:Immunoenzyme Techniques; D014116:Toxicology / Q000379:methods
D014151:Anti-Anxiety Agents; D004187:Disaccharides; D005965:Glucuronates; D001569:Benzodiazepines; D004949:Estazolam; C032318:alpha-hydroxyalprazolam; C059554:O-(glucuronic acid 2-sulfate)-(1--4)-O-(2,5)-anhydromannitol 6-sulfate; C039668:adinazolam; C050132:mono-N-demethyladinazolam; D000525:Alprazolam