High throughput quantitative analysis of the β-lyase sulfur mustard metabolite, 1,1′-sulfonylbis[2-(methylsulfinyl)ethane] in urine via high performance liquid chromatography tandem mass spectrometry

•An HPLC-MS/MS analysis for the determination of exposure to sulfur mustard (HD).•A high-throughput method with a 2.0 min cycle time and 96 well plate format.•It employs oxidative conversion of the β-lyase metabolites to SBMSE using H2O2.

Sulfur Mustard (HD) has a 100 year history of use as a chemical warfare agent and recent events in the Middle East are causing it to once again be a potential concern. We report a new high-throughput method for the determination of HD exposure by the analysis of the β-lyase metabolite 1,1′-sulfonylbis[2-(methylsulfinyl)ethane] (SBMSE) in human urine. This method features a hydrogen peroxide (H2O2) oxidative conversion of the β-lyase metabolites to SBMSE, followed by sample extraction and concentration using solid phase extraction in 96-well plate format. Subsequent high performance liquid chromatography tandem mass spectrometry (HPLC-MS/MS) analysis gave linear quantitation over a calibration range of 0.1-100 ng/mL, with a method detection limit of 0.03 ng/mL. Liquid chromatographic separation was achieved using a hydrophilic interaction liquid chromatography (HILIC) column with an analyte retention time of 0.9 min and method time of 1.5 min (cycle time = 2.0 min). Users of this method could prepare and analyze approximately 650 samples in 24 h which would be important for an emergency response.