title: Development of a liquid chromatography-multiple reaction monitoring procedure for concurrent verification of exposure to different forms of mustard agents.
abstract: A novel liquid chromatography-multiple reaction monitoring (LC-MRM) procedure has been developed for retrospective diagnosis of exposure to different forms of mustard agents. This concise method is able to validate prior exposure to nitrogen mustards (HN-1, HN-2, and HN-3) or sulfur mustard (HD) in a single run, which significantly reduces analysis time compared to separate runs to screen for different mustards' biomarkers based on tandem mass spectrometry. Belonging to one of the more toxic classes of chemical warfare agents, these potent vesicants bind covalently to the cysteine-34 residue of human serum albumin. This results in the formation of stable adducts whose identities were confirmed by a de novo sequencing bioinformatics software package. Our developed technique tracks these albumin-derived adduct biomarkers in blood samples which persist in vitro following exposure, enabling a detection limit of 200 nM of HN-1, 100 nM of HN-2, 200 nM of HN-3, or 50 nM of HD in human blood. The CWA-adducts formed in blood samples can be conveniently and sensitively analyzed by this MRM technique to allow rapid and reliable screening.
authors.affiliation: Defence Medical and Environmental Research Institute, DSO National Laboratories, 20 Science Park Drive, Singapore 118230. ythonghi@dso.org.sg
authors.forename: Thong-Hiang
authors.initials: TH
authors.lastname: Yeo
authors.forename: Mer-Lin
authors.initials: ML
authors.lastname: Ho
authors.forename: Weng-Keong
authors.initials: WK
authors.lastname: Loke
mesh_terms: D000478:Alkylation; D015415:Biomarkers / Q000097:blood; D002846:Chromatography, Affinity; D002853:Chromatography, Liquid / Q000379:methods*; D019295:Computational Biology / Q000379:methods; D004781:Environmental Exposure / Q000032:analysis; D004784:Environmental Monitoring / Q000379:methods*; D006801:Humans; D008466:Mechlorethamine / Q000032:analysis / Q000378:metabolism; D009150:Mustard Compounds / Q000032:analysis* / Q000378:metabolism; D009151:Mustard Gas / Q000032:analysis / Q000378:metabolism; D009588:Nitrogen Mustard Compounds / Q000032:analysis / Q000378:metabolism; D015203:Reproducibility of Results; D012709:Serum Albumin / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D012984:Software; D052616:Solid Phase Extraction; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D015415:Biomarkers; D009150:Mustard Compounds; D009588:Nitrogen Mustard Compounds; D012709:Serum Albumin; D008466:Mechlorethamine; C121482:bis(2-chloroethyl)ethylamine; C009802:2,2',2''-trichlorotriethylamine; D009151:Mustard Gas