title: Detection of benzodiazepines in hair using ELISA and LC-ESI-MS-MS.
abstract: This study was designed to validate an enzyme-linked immunosorbent assay (ELISA) and liquid chromatography-tandem mass spectrometry (LC-MS-MS) method for the detection of nine benzodiazepines in hair. Sixteen hair case samples were tested from drug-related deaths where a positive benzodiazepine blood result was obtained. The case samples were decontaminated with 0.1% sodium dodecyl sulfate, distilled water, and dichloromethane. For ELISA analysis, the samples were extracted by incubation in monobasic phosphate buffer for 1 h and then neutralized with dibasic phosphate buffer. They were diluted 1:5 with phosphate buffer saline (PBS) prior to analysis. For LC-MS-MS, the samples were incubated overnight in methanol/25% ammonium hydroxide (20:1). The benzodiazepines were extracted by solid phase. Thirteen samples were confirmed positive by LC-MS-MS. The benzodiazepines detected included diazepam, nordiazepam, temazepam, oxazepam, nitrazepam, and lorazepam. Using a cut-off concentration of 0.1 ng/mg oxazepam, the Immunalysis Benzodiazepine Microplate ELISA demonstrated a sensitivity and specificity of 100% and 81%, respectively, compared with LC-MS-MS results.
authors.affiliation: Forensic Medicine and Science Department, University of Glasgow, University Place, Glasgow, G12 8QQ, Scotland. ei_miller2003@yahoo.co.uk
authors.forename: Eleanor I
authors.initials: EI
authors.lastname: Miller
authors.forename: Fiona M
authors.initials: FM
authors.lastname: Wylie
authors.forename: John S
authors.initials: JS
authors.lastname: Oliver
mesh_terms: D001569:Benzodiazepines / Q000032:analysis* / Q000097:blood; D002853:Chromatography, Liquid / Q000379:methods; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D006197:Hair / Q000737:chemistry*; D006801:Humans; D006993:Hypnotics and Sedatives / Q000032:analysis* / Q000097:blood; D015203:Reproducibility of Results; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods; D015813:Substance Abuse Detection
chemical_list: D006993:Hypnotics and Sedatives; D001569:Benzodiazepines