Medical Aspects of Chemical Warfare (2008) - The Black Vault

Medical DiagnosticsLC-MS-MS method that can analyze the compounddirectly. 98 Other possible urinary analytes are an imidazolederivative formed from sulfur mustard’s reactionwith protein histidine residues 99 and sulfur mustardadducts to metallothionien. 100Application to Human ExposureVycudilik analyzed urine samples from two casualtiesof the Iran-Iraq War who were brought to a hospitalin Vienna, Austria, for treatment of suspected exposureto sulfur mustard. 81 The exposure was believed tohave occurred 1 week prior to their arrival in Vienna.No clinical description of the patients’ injuries wasprovided in the report. The concentration of sulfurmustard found in their urine samples using GC-MSwas approximately 1.0 ng/mL and 1.5 ng/mL. Additionalurine samples were obtained from the patientsseveral days after admission to the hospital. Analysisusing the same method produced negative results forall samples.Vycudilik also analyzed urine samples obtainedfrom 12 Iran-Iraq War casualties. 82 The only clinicaldescription of the patients was the observation thatthey had severe skin lesions resulting from an allegedmustard gas attack. Urine samples from six of thepatients produced positive results for sulfur mustard.Concentrations found ranged from 1 to 30 ng/mL. Themethod could not distinguish between sulfur mustardor its hydroxyethyl metabolites present in the urinesamples.Wils et al examined a large number of urine samplesfor TDG concentrations. The samples were obtainedfrom Iranian casualties of the Iran-Iraq War transportedto western European hospitals in Ghent and Utrechtfor treatment. 83,84 The majority of the urine samplesEXHIBIT 22-1SAMPLE PREPARATION METHODS FOR THE Gas Chromatographic/Mass Spectrometric/MassSpectrometric ANALYSIS OF THE SULFUR MUSTARD URINARY β-LYASEMETABOLITESProcedure of Black et al:• Add the internal standard deuterated 1,1’-sulfonylbis[2-(methylsulfinyl)ethane] to 1 mL of urine.• Add 0.4 mL of titanium trichloride.• Incubate sample at 40°C overnight (16 hours).• Filter solution through a preconditioned C 8Bond Elut solid-phase extraction cartridge.• Wash cartridge with water followed by a methanol and water mixture.• Allow cartridge to dry.• Elute analytes with acetone.• Evaporate to dryness under nitrogen and dissolve in toluene.• Analyze using GC-MS-MS with ammonia chemical ionization. 1Procedure of Young et al:• Place 0.5 mL of urine into a 15 mL tube.• Add the internal standard 13 C-1,1’-sulfonylbis[2-(methythio)ethane] to the urine.• Add 1 mL of titanium trichloride.• Incubate sample at 75°C for 1 hour.• Add 2 mL of 6N sodium hydroxide and mix.• Centrifuge samples for 5 minutes.• Pour supernatant into a Chem Elut column.• Elute analytes with 16 mL of dichloromethane and acetonitrile mixture.• Evaporate to dryness under nitrogen and dissolve in toluene.• Analyze using GC-MS-MS with isobutane chemical ionization. 2GC: gas chromatographyMS: mass spectrometryData sources: (1) Black RM, Clarke RJ, Read RW. Analysis of 1,1’-sulphonylbis[2-(methylsulphinyl)ethane] and 1-methylsulphinyl-2-[2-(methylthio)ethylsulphonyl]ethane, metabolites of sulphur mustard, in urine using gas chromatography-mass spectrometry.J Chromatogr. 1991;558:405–414. (2) Young CL, Ash D, Driskell WJ, et al. A rapid, sensitive method for the quantitation of specificmetabolites of sulfur mustard in human urine using isotope-dilution gas chromatography-tandem mass spectrometry. J Anal Toxicol.2004;28:339–345.713