title: Monitoring urinary metabolites resulting from sulfur mustard exposure in rabbits, using highly sensitive isotope-dilution gas chromatography-mass spectrometry.
abstract: A highly sensitive method for the determination of sulfur mustard (SM) metabolites thiodiglycol (TDG) and thiodiglycol sulfoxide (TDGO) in urine was established and validated using isotope-dilution negative-ion chemical ionization (NICI) gas chromatography-mass spectrometry (GC-MS). TDGO in the samples was reduced with TiCl3, and then determined together with TDG as a single analyte. The sample preparation procedures, including two solid-phase-extraction (SPE) clean-up steps, were optimized to improve the sensitivity of the method. The limits of detection (LOD) for both TDG and TDG plus TDGO (TDG + TDGO) were 0.1 ng mL(-1), and the limits of quantitation (LOQ) for both were 0.3 ng mL(-1). The method was used in a rabbit cutaneous SM exposure model. Domestic rabbits were exposed to neat liquid SM at three dosage levels (0.02, 0.05, and 0.15 LD50), and the urinary excretion of four species of hydrolysis metabolites, namely free TDG, free plus conjugated TDG (total TDG), free TDG + TDGO, and free plus conjugated TDG + TDGO (total TDG + TDGO), was evaluated to investigate the metabolic processes. The total urinary excretion profiles of the metabolites, including the peak time, time window, and dose-response and time-response relationships, were clarified. The results revealed that the concentrations of TDG and TDG + TDGO in the urine increased quickly and then decreased rapidly in the first two days after SM exposure. The cumulative amount of total TDG + TDGO excreted in urine during the first five days accounted for 0.5-1% of the applied dose of SM. It is also concluded that TDG and TDGO in urine existed mainly in free form, the levels of glucuronide and of sulfate conjugates of TDG or TDGO were very low, and most hydrolysis metabolites were present in the oxidized form (TDGO). The study indicates that the abnormal increase of TDG and TDGO excretion levels can be used as a diagnostic indicator and establishes a reference time-window for retrospective analysis and sampling after SM exposure.
authors.affiliation: State Key Laboratory of Toxicology and Medical Countermeasures, and Laboratory of Toxicant Analysis, Institute of Pharmacology and Toxicology, Academy of Military Medical Sciences, 27 Taiping Road, Haidian District, Beijing, 100850, China.
authors.forename: Zhiyong
authors.initials: Z
authors.lastname: Nie
authors.forename: Yajiao
authors.initials: Y
authors.lastname: Zhang
authors.forename: Jia
authors.initials: J
authors.lastname: Chen
authors.forename: Ying
authors.initials: Y
authors.lastname: Lin
authors.forename: Bidong
authors.initials: B
authors.lastname: Wu
authors.forename: Yuan
authors.initials: Y
authors.lastname: Dong
authors.forename: Jianlin
authors.initials: J
authors.lastname: Feng
authors.forename: Qin
authors.initials: Q
authors.lastname: Liu
authors.forename: Jianwei
authors.initials: J
authors.lastname: Xie
mesh_terms: D000279:Administration, Cutaneous; D000818:Animals; D001711:Biotransformation; D002619:Chemical Warfare Agents / Q000378:metabolism / Q000633:toxicity*; D003879:Dermatologic Agents / Q000378:metabolism / Q000633:toxicity*; D004305:Dose-Response Relationship, Drug; D008401:Gas Chromatography-Mass Spectrometry / Q000379:methods; D007201:Indicator Dilution Techniques; D008297:Male; D009151:Mustard Gas / Q000378:metabolism / Q000633:toxicity*; D010084:Oxidation-Reduction; D011817:Rabbits; D012867:Skin / Q000098:blood supply / Q000187:drug effects / Q000378:metabolism; D052616:Solid Phase Extraction; D013438:Sulfhydryl Compounds / Q000652:urine*; D013454:Sulfoxides / Q000652:urine*; D014025:Titanium / Q000737:chemistry
chemical_list: D002619:Chemical Warfare Agents; D003879:Dermatologic Agents; D013438:Sulfhydryl Compounds; D013454:Sulfoxides; C073450:2,2'-sulfinylbisethanol; C047519:2,2'-thiodiethanol; D014025:Titanium; C039460:titanium trichloride; D009151:Mustard Gas