An Enhanced Throughput Method for Quantification of Sulfur Mustard Adducts to Human Serum Albumin via Isotope Dilution Tandem Mass Spectrometry
Published Date:2014 Jan-Feb
Source:J Anal Toxicol. 38(1):8-15.
High-Throughput Screening Assays
Sensitivity And Specificity
Solid Phase Extraction
Tandem Mass Spectrometry
Pubmed Central ID:PMC4539155
Funding:CC999999/Intramural CDC HHS/United States
Description:Here, we report an enhanced throughput method for the diagnosis of human exposure to sulfur mustard. A hydroxyethylthioethyl (HETE) ester-adducted tripeptide, produced by pronase digestion of human serum albumin, was selected as the quantitative exposure biomarker. Cibacron Blue enrichment was developed from an established cartridge method into a 96-well plate format, increasing throughput and ruggedness. This new method decreased sample volume 2.5-fold. Addition of a precipitation and solid-phase extraction concentration step increased the sensitivity of the method. With the conversion to a 96-well plate and optimization of chromatography, the method resulted in a 3-fold decrease in analysis time. Inclusion of a confirmation ion has increased specificity. The method was found to be linear between 0.050 and 50 µM sulfur mustard exposure with a precision for both quality control samples of ≤6.5% relative standard deviation and an accuracy of >96%. The limit of detection (3So) was calculated to be ∼0.0048 µM, an exposure value similar to that of the HETE-albumin adduct method first described by Noort and co-workers (Noort et al., 1999; Noort el al., 2004) which used protein precipitation to isolate albumin. A convenience set of 124 plasma samples from healthy unexposed individuals was analyzed using this method to assess background levels of exposure to sulfur mustard; no positive results were detected.
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