Simultaneous Quantification of Triazolam and its Metabolites in Human Urine by Liquid Chromatography-Tandem Mass Spectrometry

Abstract

A simple, simultaneous, sensitive and selective liquid chromatography-tandem mass spectrometry (LC-MS-MS) method for the determination of triazolam and its metabolites, α-hydroxytriazolam (α-OHTRZ) and 4-hydroxytriazolam (4-OHTRZ), in human urine was developed and validated. Triazolam-d4 was used as the internal standard (IS). This analysis was carried out on a Thermo(®) C(18) column, and the mobile phase was composed of acetonitrile/H(2)O/formic acid (35:65:0.2, v/v/v). Detection was performed on a triple-quadrupole tandem MS using positive ion mode electrospray ionization, and quantification was performed by multiple reaction monitoring mode. The MS-MS ion transitions monitored were m/z 343.1 → 308.3, 359.0 → 331.0, 359.0 → 111.2, and 347.0 → 312.0 for triazolam, α-OHTRZ, 4-OHTRZ, and triazolam-d(4), respectively. The lower limits of quantification of the analytical method were 0.5 ng/mL for triazolam, 5 ng/mL for α-OHTRZ, and 0.5 ng/mL for 4-OHTRZ. The within- and between-run precisions were less than 15%, and accuracy was -12.33% to 9.76%. The method was proved to be accurate and specific, and it was applied to a urinary excretion study of triazolam in healthy Chinese volunteers.

A drug rape case involving triazolam detected in hair and urine

“…Furthermore, controlled studied of single dose intake in humans such as the study on ketamine by Xiang et al [13] would be interesting to perform with triazolam in order to obtain more knowledge on levels in hair and the influence of sweat and sebum on segments. The detection of α-hydroxytriazolam in urine concurs with other studies of triazolam metabolism [14,15]. These have reported that α-hydroxytriazolam is the major metabolite excreted in urine within the first 12 h. However, they also observed low amounts of it 30 to 40 h after intake.…”