title: An enhanced throughput method for quantification of sulfur mustard adducts to human serum albumin via isotope dilution tandem mass spectrometry.
abstract: 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.
authors.affiliation: 1Emergency Response Branch, Division of Laboratory Sciences, National Center for Environmental Health, Centers for Disease Control and Prevention, 4770 Buford Highway, NE, Atlanta, GA 30341, USA.
authors.forename: Tracy M
authors.initials: TM
authors.lastname: Andacht
authors.forename: Brooke G
authors.initials: BG
authors.lastname: Pantazides
authors.forename: Brian S
authors.initials: BS
authors.lastname: Crow
authors.forename: Alex
authors.initials: A
authors.lastname: Fidder
authors.forename: Daan
authors.initials: D
authors.lastname: Noort
authors.forename: Jerry D
authors.initials: JD
authors.lastname: Thomas
authors.forename: Thomas A
authors.initials: TA
authors.lastname: Blake
authors.forename: Rudolph C
authors.initials: RC
authors.lastname: Johnson
mesh_terms: D015415:Biomarkers / Q000097:blood; D002138:Calibration; D004781:Environmental Exposure / Q000032:analysis*; D057166:High-Throughput Screening Assays / Q000379:methods*; D006801:Humans; D009151:Mustard Gas / Q000633:toxicity*; D040901:Proteomics; D012189:Retrospective Studies; D012680:Sensitivity and Specificity; D012709:Serum Albumin / Q000737:chemistry*; D052616:Solid Phase Extraction; D013048:Specimen Handling; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D015415:Biomarkers; D012709:Serum Albumin; D009151:Mustard Gas