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

• Published in: Journal of analytical toxicology 2011-04, Vol.35 (3), p.170-175
• Main Authors: Pan, Ryh-Nan, Hsiong, Cheng-Huei, Huang, Pei-Wei, Tsai, Wei-Shiuan, Li, Wen-Yi, Pao, Li-Heng
• Format: Article
• Language: English
• Subjects: Adjuvants, Anesthesia - chemistry; Adjuvants, Anesthesia - urine; Adult; Analysis; Biological and medical sciences; Chromatography, Liquid; General pharmacology; Humans; Male; Medical sciences; Pharmacology. Drug treatments; Substance Abuse Detection - methods; Tandem Mass Spectrometry; Triazolam - analogs & derivatives; Triazolam - chemistry; Triazolam - urine
• ISSN: 0146-4760; 1945-2403
• DOI: 10.1093/anatox/35.3.170

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® C18 column, and the mobile phase was composed of acetonitrile/H2O/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-d4, 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.