Analysis of Urinary Metabolites of Sulfur Mustard in Two Individuals after Accidental Exposure

Abstract

]In July 2004, two individuals developed blisters after the destruction of a WWl-era munition. To determine the causative agent, urine samples were collected from both the highly blistered patient (patient 1; 6.5% of total body surface area) and patient 2, who had only one small blister. Their urine was analyzed for metabolites of known vesicants including sulfur mustard (HD), Lewisite (L1), and nitrogen mustards. The urine samples only tested positive for metabolites of HD. Additional metabolites were measured to confirm the exposure of sulfur mustard agent HD, including thiodiglycol (TDG), TDG-sulfoxide, and the bismercapturate of mustard sulfone. On day 2 after the exposure, patient I had a lS-lyase metabolite level of 41 ng/mL, and patient 2 had a level of 2.6 ng/mt. Detectable levels of the ~-Iyase metabolite were observed in patient 1 for 11 days and in patient 2 for 7 days. Levels of TDG and both TDG and its sulfoxide measured together in the urine of patient 1 were found to be 24 ng/mL and 50 ng/mL, respectively, on day 2. The bis-mercapturate of mustard sulfone was detected in patient 1 (3.1 ng/mL) on day 2 but was not detected in samples taken on subsequent days.The metabolism of HD has been studied extensively in rats (3,4). Research has identified several urinary metabolites, including the free and conjugated forms of the simple hydrolysis product thiodiglycol (TDG) and TDG-sulfoxide (TDG's oxidized form), the glutathione-derived metabolite 1,1'-sul- fonylbis[2-S-(N-acetylcysteinyl)ethane], and the I~-lyase metabolites 1,1'-sulfonylbis[2-(methylsulfinyl)ethane] (SBMSE) and 1-methylsulfinyl-2-[2-(methylthio)-ethylsulfonyl]ethane (MSMTESE). Although 1,]'-sulfonylbis[2-(methylthio)ethane] (SBMTE)was not identified in rat urine, it is a likely precursor to the I~-lyase metabolites that were identified (3-8).Recently, two individuals were thought to have been exposed to HD accidentally during the destruction of a WWI-era shell (9,10). The extent of blistering between the two individuals varied. One individual (patient 1) had blisters on an estimated 6.5% of his body including his hands and forearms; the second individual (patient 2) had only one small blister (9,10). Urine samples from both individuals were analyzed to confirm that the symptoms were caused by HD exposure.

Accumulation of intact sulfur mustard in adipose tissue and toxicokinetics by chemical conversion and isotope-dilution liquid chromatography–tandem mass spectrometry

“…A series of oxidation, hydrolysis or β-lyase-free metabolites and covalent adducts with macromolecules of SM have been identified in urine (Barr et al 2008;Black et al 1992b;Black and Read 1995;Davison et al 1961;Fidder et al 1996b;Read and Black 2004;Sandelowsky et al 1992;Zhang et al 2014) and blood Black et al 1997a, b;Fidder et al , 1996aNoort et al 1997;Sandelowsky et al 1992;van der Schans et al 1994) after SM exposure. The excretion or toxicokinetic profiles of these metabolites and adducts were studied to get an insight into the mechanism of toxicity and metabolism of SM.…”

Development and validation of efficient stable isotope dilution LC–HESI–MS/MS method for the verification of β-lyase metabolites in human urine after sulfur mustard exposure

“…LOQs (Formulas (3) and (4)) were 10 ng/ml and 11 ng/ml for SBMSE and MSMTESE, respectively. In previous studies of victims after exposure [11,12], measured combined amount of SBMSE and MSMTESE from urine have been 42-57 ng/ml between 1 and 3 days after exposure. These results have been measured after reduction and GC-MS analysis.…”

“…The metabolism of HD has been studied in vivo in rats [10]. Some of these metabolites were identified in human subjects accidentally or deliberately exposed to sulfur mustard [11][12][13]. These were the hydrolysis products thiodiglycol (TDG) and thiodiglygol sulfoxide (TDGO) but also the 1,1 -sulfonylbis[2-S-(N-acetylcysteinyl)ethane (SBSNAE) and the ␤-lyase metabolites 1,1 -sulfonylbis-[2-(methylsulfinyl)ethane] (SBMSE) and…”

“…These are regarded as unequivocal biomarkers of exposure to sulfur mustard [19,20]. SBSNAE, also derived from an initial reaction with glutathione, appears to be a minor metabolite in humans based on the analysis of samples after three accidental exposed victims [12,21]. Accurate analytic methods using both gas chromatography-tandem mass spectrometry (GC-MS/MS) [17,18] and liquid chromatography-tandem mass spectrometry (LC-MS/MS) [20] have been published.…”

History and perspectives of bioanalytical methods for chemical warfare agent detection

“…Adducts were identified in vitro, with nucleophilic amino acid residues on haemoglobin (Nterminal valine, histidine, aspartic and glutamic acids) [30,31], albumin (cysteine) [32], and with DNA [33]. Following the development of analytical methods, based on GC-MS, GC-MS-MS, LC-MS-MS, and immunoassay (DNA), all of these adducts have been detected retrospectively in stored blood samples from Iranian casualties [34,35], and/or in samples from accidental human exposures [35,36].…”