title: Gas chromatographic-mass spectrometric analysis of sulfur mustard-plasma protein adducts: validation and use in a rat inhalation model.
abstract: Sulfur mustard (HD) is an alkylating agent that reacts rapidly with macromolecular targets resulting in the formation of stable adducts providing depots for markers of exposure. The purpose of this study was to validate an analytical procedure for detection of HD-plasma protein adducts and to establish the utility of the method in an HD rat inhalation study. Calibration curves were prepared in human and rat plasma at six levels of HD (12.5 to 400 nM). Correlation coefficients for the mean data were 0.9987 for human and 0.9992 for rat plasma. The percent coefficient of variation (%CV) derived from the mean concentration data ranged from 0.53 to 14.1% in human (n = 5) and 0.57 to 10.63% in rat (n = 6) plasma. Intraday and interday precision and accuracy studies were conducted at three concentration levels (25, 150, 300 nM) to represent low, medium, and high concentrations of HD relative to those employed in the calibration curve. Precision and accuracy were assessed by determining %CV and % error, respectively. For intra- and interday studies, the %CVs and absolute % errors were less than 15%. The limits of quantitation were 20.88 nM for human and 16.73 nM for rat plasma. In animal studies, rats received nebulized HD at six doses. The data indicate a dose-dependent relationship between maximal plasma concentrations and dose administered (R(2) = 0.9728). Results from this study indicate an accurate, precise, and sensitive method. The method was useful in determining plasma protein adduct formation in a rat inhalation model.
authors.affiliation: Analytical Toxicology Division, US Army Medical Research Institute of Chemical Defense, 3100 Ricketts Point Road, Aberdeen Proving Ground, Maryland 21010-5400, USA. benedict.capacio@us.army.mil
authors.forename: Benedict R
authors.initials: BR
authors.lastname: Capacio
authors.forename: J Richard
authors.initials: JR
authors.lastname: Smith
authors.forename: Richard J
authors.initials: RJ
authors.lastname: Lawrence
authors.forename: Brian L
authors.initials: BL
authors.lastname: Boyd
authors.forename: Alicia M
authors.initials: AM
authors.lastname: Witriol
authors.forename: Michele L
authors.initials: ML
authors.lastname: Conti
authors.forename: Jennifer L
authors.initials: JL
authors.lastname: Collins
authors.forename: Alfred M
authors.initials: AM
authors.lastname: Sciuto
mesh_terms: D000280:Administration, Inhalation; D000478:Alkylation; D000818:Animals; D001565:Benzoates / Q000737:chemistry; D015415:Biomarkers / Q000097:blood; D001798:Blood Proteins / Q000737:chemistry / Q000378:metabolism*; D002138:Calibration; D004781:Environmental Exposure / Q000032:analysis; D004784:Environmental Monitoring / Q000379:methods*; D008401:Gas Chromatography-Mass Spectrometry / Q000379:methods*; D006801:Humans; D008297:Male; D009151:Mustard Gas / Q000008:administration & dosage / Q000032:analysis* / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D015203:Reproducibility of Results; D012972:Sodium Hydroxide / Q000737:chemistry; D013438:Sulfhydryl Compounds / Q000032:analysis / Q000737:chemistry; D005464:Fluorobenzenes; D005466:Fluorocarbons
chemical_list: D001565:Benzoates; D015415:Biomarkers; D001798:Blood Proteins; D009151:Mustard Gas; D012972:Sodium Hydroxide; D013438:Sulfhydryl Compounds; C014737:pentafluorobenzoyl chloride; D005464:Fluorobenzenes; D005466:Fluorocarbons; C047519:2,2'-thiodiethanol