title: Liquid chromatography-tandem mass spectrometry method for the determination of thiosulfate in human blood and urine as an indicator of hydrogen sulfide poisoning.
abstract: Being a stable metabolite of hydrogen sulfide, thiosulfate has been utilized as an index for hydrogen sulfide poisoning (HSP). Thiosulfate analysis is mainly performed using gas chromatography/mass spectrometry (GC-MS) due to its high sensitivity and specificity. The GC-MS analysis requires two-step derivatizations of thiosulfate, and the derivative is not stable in solution as it has a disulfide moiety. To resolve this stability issue, we developed a novel analytical method using liquid chromatography-tandem mass spectrometry (LC-MS/MS) for monitoring the pentafluorobenzyl derivative of thiosulfate (the first reaction product of the GC-MS method) in this study. The established method exhibited high reproducibility despite being a more simplified and rapid procedure compare to the GC-MS method. Phenyl 4-hydroxybenzoate was used as an internal standard because 1,3,5-tribromobenzene which had been used in the GC-MS method was not suitable compound for LC-MS/MS with Electrospray ionization (ESI) negative detection. The linear regression of the peak area ratios versus concentrations was fitted over the concentration ranges of 0.5-250μM and 0.25-250μM in blood and urine, respectively. The validation results satisfied the acceptance criteria for intra- and inter-day accuracy and precision. Blood and urine samples from 12 suspected HSP cases were tested using this method. The thiosulfate concentration detected in the sample coincided well with that determined at the scene of each HSP accident.
authors.affiliation: Department of Legal Medicine, Asahikawa Medical University, 2-1-1-1 Midorigaoka-Higashi, Asahikawa 078-8510, Japan.
authors.forename: Chikatoshi
authors.initials: C
authors.lastname: Maseda
authors.affiliation: Tokyo Medical Examiner's Office, Tokyo Metropolitan Government, 4-21-18 Otsuka, Bunkyo-ku, Tokyo 112-0012, Japan.
authors.forename: Akira
authors.initials: A
authors.lastname: Hayakawa
authors.affiliation: Department of Legal Medicine, Asahikawa Medical University, 2-1-1-1 Midorigaoka-Higashi, Asahikawa 078-8510, Japan. Electronic address: k-okuda@asahikawa-med.ac.jp.
authors.forename: Katsuhiro
authors.initials: K
authors.lastname: Okuda
authors.affiliation: Department of Legal Medicine, Asahikawa Medical University, 2-1-1-1 Midorigaoka-Higashi, Asahikawa 078-8510, Japan.
authors.forename: Masaru
authors.initials: M
authors.lastname: Asari
authors.affiliation: Department of Legal Medicine, Asahikawa Medical University, 2-1-1-1 Midorigaoka-Higashi, Asahikawa 078-8510, Japan.
authors.forename: Hiroki
authors.initials: H
authors.lastname: Tanaka
authors.affiliation: Department of Legal Medicine, Asahikawa Medical University, 2-1-1-1 Midorigaoka-Higashi, Asahikawa 078-8510, Japan.
authors.forename: Hiromi
authors.initials: H
authors.lastname: Yamada
authors.affiliation: Faculty of Health Sciences, Hokkaido University Graduate School of Medicine, N 12, W 5, Kita-ku, Sapporo 060-0812, Japan.
authors.forename: Shigeki
authors.initials: S
authors.lastname: Jin
authors.affiliation: Technical Support Department, Faculty of Medicine, Hokkaido University, N 15, W 7, Kita-ku, Sapporo 060-8638, Japan.
authors.forename: Kie
authors.initials: K
authors.lastname: Horioka
authors.affiliation: Department of Forensic Medicine, Hokkaido University Graduate School of Medicine, N 15, W 7, Kita-ku, Sapporo 060-8638, Japan.
authors.forename: Kotaro
authors.initials: K
authors.lastname: Matoba
authors.affiliation: Department of Legal Medicine, Asahikawa Medical University, 2-1-1-1 Midorigaoka-Higashi, Asahikawa 078-8510, Japan.
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Shiono
authors.affiliation: Department of Clinical Pharmacology and Therapeutics, Kyoto University Hospital, 54 Shogoin-Kawahara-cho, Sakyo-ku, Kyoto 606-8507, Japan.
authors.forename: Kazuo
authors.initials: K
authors.lastname: Matsubara
authors.affiliation: Department of Legal Medicine, Asahikawa Medical University, 2-1-1-1 Midorigaoka-Higashi, Asahikawa 078-8510, Japan.
authors.forename: Keiko
authors.initials: K
authors.lastname: Shimizu
mesh_terms: D002853:Chromatography, Liquid / Q000379:methods*; D053593:Forensic Toxicology; D006801:Humans; D006862:Hydrogen Sulfide / Q000506:poisoning*; D036301:Qualitative Research; D053719:Tandem Mass Spectrometry / Q000379:methods*; D013885:Thiosulfates / Q000097:blood* / Q000652:urine*
chemical_list: D013885:Thiosulfates; D006862:Hydrogen Sulfide
keywords: Hydrogen sulfide; LC-MS/MS; Quantitative determination; Thiosulfate