Simultaneous quantification of seven plasma metabolites of sulfur mustard by ultra high performance liquid chromatography–tandem mass spectrometry

Sulfur mustard (SM) is a hazardous chemical warfare agent that has been used in several military conflicts. SM is also considered as a major threat to civilians because of its existing stockpiles and easy production. Analysis of exposure biomarkers in biological samples collected from suspected victims is a useful tool for early diagnosis of SM poisoning. In this study, a sensitive and rapid quantitative method with ultra high performance liquid chromatography–tandem mass spectrometry (UPLC–MS/MS) was developed for simultaneous determination of seven SM plasma biomarkers, including its oxidative, hydrolysis and β-lyase metabolites. A simple one-step protein precipitation with acetonitrile–methanol (4:1) was used for sample preparation. A full validation was conducted with respect to specificity, linearity, recovery, matrix effect, precision, accuracy and stability. The lower limits of quantification for the seven metabolites ranged from 0.01 μg L−1 to 5 μg L−1. The intraday relative standard deviation was less than 7.0%, and the interday deviation was less than 9.1%. The recoveries varied in the range from 82.8% to 118%. This method has been successfully applied to a toxicokinetic study for obtaining the plasma profiles of seven metabolites in SM-exposed rats, following a single subcutaneous dose of 3.3 mg kg−1. All the targeted compounds were detected in rat plasma. bis-β-Chloroethyl sulfoxide (SMO), thiodiglycol (TDG), thiodiglycol sulfoxide (TDGO), 1,1′-sulfonylbis-[2-S-(N-acetylcysteinyl)ethane (SBSNAE), 1,1′-sulfonylbis-[2-(methylsulfinyl)ethane] (SBMSE) and 1-methylsulfinyl-2-[2-(methylthio)ethylsulfonyl]ethane (MSMTESE) were found to be the major metabolites in rat plasma. The time windows for the detection of these metabolites were varied in the range of 5 min to 48 h after exposure. The method provides a useful tool for short-term diagnosis of SM poisoning.

► A novel UPLC–MS/MS method was established and fully validated.
► It was used for simultaneous quantitation of 7 plasma sulfur mustard biomarkers.
► The method has been proved to be simple, rapid and sensitive.
► It is suitable for early monitoring sulfur mustard poisoning.
► The plasma profiles of these biomarkers in rats were reported for the first time.