Urinalysis of α-Hydroxyalprazolam, α-Hydroxytriazolam, and Other Benzodiazepine Compounds by GC/EIMS

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Abstract
This procedure was developed as an overall benzodiazepine confirmation scheme and includes the detection of the most important urinary analytes encountered by clinical toxicology laboratories in North America: α-hydroxyalprazolam, α-hydroxytriazolam, 2-hydroxyethylflurazepam, oxazepam, temazepam, and Iorazepam. Desmethyldiazepam (nordiazepam) was not targeted because it Is metabolized to oxazepam. This procedure takes advantage of β-glucuronidase hydrolysis for analysis of intact benzodiazepine molecules, oxazepam-2H5 as an internal standard, a newly developed extraction solvent, and a silylating moiety that may be more sensitive than trimethylsilyl (-TMS) derivatives, the tertbutyldimethylsilyl (-TBDMS) derivative. For all compounds the extraction efficiency was greater than 90% and the limit of quantitation (LOQ at a S/N of 10) was less than 10 ng/mL. Coefficients of variation for a 200-ng/mL control were less than 5% and less than or equal to 11% for within-run and between-run trials, respectively. Of 13 human specimens screened by EMIT® and most with self-reported histories, α-hydroxyalprazolam was found in seven (range 49–1264 ng/mL), oxazepam was found in five (72–3897 ng/mL), and lorazepam (476 ng/mL), 2-hydroxyethylflurazepam (2301 ng/mL), and α-hydroxytriazolam (106 ng/mL) in one each.

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