Analytical methodology for the detection of benzodiazepine consumption in opioid-dependent subjects

Benzodiazepines are frequently abused by heroin users, but not all compounds have shown the same abuse liability. We developed an analytical method that was able to detect various benzodiazepine compounds in a single run. Enzymatically hydrolyzed urine underwent a liquid-liquid extraction procedure with chloroform/isopropanol (9:1) at pH 8-9 followed by a solid-liquid clean-up (Bond Elut TCA C18) to obtain appropriate extracts for HPLC analysis. Mobile phase composition was optimized by means of the linear solvation energy relationship methodology based on Reichardt's normalized solvatochromic parameter (E(T)N). The method was validated for the simultaneous detection and quantitation of alprazolam, alpha-hydroxyalprazolam, oxazepam, 7-aminoflunitrazepam, flunitrazepam, nordiazepam, lorazepam, and diazepam. The prevalence of benzodiazepine consumption in 229 opioid-dependent subjects on methadone-maintenance treatment was 48%. Oxazepam and nordiazepam were the benzodiazepines most frequently recoved in urine samples. The prevalence of alprazolam use (40%) was higher than that of flunitrazepam (10%).