Forensic analysis of triazolam in human tissues using capillary gas chromatography

Abstract

A reliable and sensitive method has been developed to assess the concentrations of the hypnotic drug triazolam in human tissues, including putrefied tissues. The method involves a 3-step solvent extraction, clean-up on a silica gel column and gas chromatography using a nitrogen phosphorus detector and a capillary column. Estazolam was used as an internal standard. The calibration curve was linear over the concentration range 1 ng/g-1 microgram/g and the lower limit of detection was 0.5 ng/g. A forensic study was performed on the toxicological effects of triazolam using putrefied tissues.

Detection of triazolam in skeletal remains buried for 4 years

“…As the amount of triazolam in the bone marrow samples was near the detection limit of GC/MS with NICI mode, direct subjection of the extracts to GC/MS was attempted in order to avoid contamination during the HPLC step of purification. Our extraction procedure [1] was modified so that even a trace amount of triazolam in skeletonizing remains could be selectively determined by GC/MS with NICI mode. Fewer interfering peaks were observed on the chromatogram by digesting samples with 2 M sodium hydroxide and by washing the aqueous layer with hexane twice in the 3-step solvent extraction procedure.…”

“…The method previously established for the analyses of triazolam in human solid tissues [1] was first used for the mummified tissue. The drug was extracted using a 3-step solvent extraction procedure, purified on a silica gel column, and analyzed by gas chromatography with a nitro- gen phosphorus detector.…”

“…Extraction procedure A method we previously reported was modified so that trace amounts of triazolam in decomposed tissues could be analyzed [1,3]. Bone marrow or mummified muscle samples (1 g) were mixed with 1 µl of IS solution (10 ng of estazolam) in a 30-ml centrifuge tube, and digested with 5 ml of 2 M sodium hydroxide at 100°C for 30 min.…”

“…As dosage of this drug is as low as 0.125 mg in one tablet, and the half-life is short (1.8-3.9 h), the detection and quantitation of minute amounts of triazolam in human tissues are difficult especially in cases of extensive putrefaction [1,2].…”

Gas chromatography–tandem mass spectrometry assay for the quantification of four benzodiazepines and citalopram in eleven postmortem rabbit fluids and tissues, with application to animal and human samples

“…Other benzodiazepines have also been quantified in various tissues for case reports or pharmacokinetic studies. Excepted one study of bromazepam [21] that included a fully validated assay on a large number of matrices; in other published studies, analytical validation was either lacking [22][23][24][25] or performed on blood samples [26,27] or on part of the studied matrices [28].…”