Diagnosing human exposure to sulfur mustard by measuring human serum albumin adducts via isotope dilution tandem mass spectrometry (Poster)
conference paper
Sulfur mustard agent (HD) (2,2’-dichloroethyl sulfide) is a reactive electrophile that readily alkylates aromatic nitrogen atoms, carboxyl groups, sulfides, and sulfhydryl groups on DNA and protein. Adducts to both DNA and specific proteins have been used to assess human exposure to HD. Human serum albumin (HSA) is alkylated by HD on the only reduced cysteine residue, Cys-34. HSA serves as an excellent biomarker due to its abundance, residence time in the body, and ease of purification. Here we report on improvements to the current method for assessing HD exposure via quantification of an adducted pronase digest peptide from HSA by liquid chromatography-tandem mass spectrometry (LC-MS/MS). The method has been converted to a 96-well plate format, increasing throughput and ruggedness, and decreasing sample quantity requirements. This format decreases the sample preparation time by 3-fold compared to the manual method, and requires only 200 μL of sample. A precipitation step and solid-phase extraction concentration step have been included, increasing sensitivity 10-fold. The high performance liquid chromatography (HPLC) conditions have been optimized, decreasing analysis time by 4-fold. In addition, a confirmation ion has been included to increase specificity. The method is linear between 0.1 and 100 μM, with precision for both quality control samples approximately 5%, and accuracy >90%.
TNO Identifier
460687
Source title
18th Biennial Medical Chemical Defense Bioscience Review, Baltimore Marriott Hunt Valley Inn, Hunt Valley, Maryland, USA, 20-24 May 2012
Pages
Poster 26
Files
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