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Published:
Journal of Analytical Toxicology,
ISSN 0146-4760,
Volume 32, Number 1, January/February,
pp.51-56
Development of a Liquid Chromatography– Multiple
Reaction Monitoring Procedure for Concurrent Verification of
Exposure to Different Forms of Mustard Agents
Thong-Hiang Yeo, Mer-Lin Ho, and Weng-Keong Loke
Defence Medical and Environmental Research Institute, DSO National
Laboratories, 20 Science Park Drive, Singapore 118230
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 200nM of HN-1, 100nM
of HN-2, 200nM of HN-3, or 50nM 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.
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