title: Monitoring exposure to acrylonitrile using adducts with N-terminal valine in hemoglobin.
abstract: Human exposure to acrylonitrile (ACN), a carcinogen in rats, may occur in industrial settings, through waste water and tobacco smoke. ACN is an electrophilic compound and binds covalently to nucleophilic sites in macromolecules. Measurements of adducts with hemoglobin could be utilized for improved exposure assessments. In this study, a method for quantification of N-(2-cyanoethyl)valine (CEVal), the product of reaction of ACN with N-terminal valine in hemoglobin has been developed. The method is based on the N-alkyl Edman procedure, which involves derivatization of the globin with pentafluorophenyl isothiocyanate and gas chromatographic-mass spectrometric analysis of the resulting thiohydantoin. An internal standard was prepared by reacting valylglycylglycine with [2H3]ACN, spiked with [14C]ACN to a known sp. act. Levels of CEVal were measured in globin from rats exposed to 3-300 p.p.m. ACN in drinking water for 105 days and from humans (four smokers and four non-smokers). CEVal was detected at all exposure levels in the drinking water study. The relationship between adduct level and water concentration was linear at concentrations of 10 p.p.m. (corresponding to an average daily uptake of c. 0.74 mg ACN/kg body wt during the 65 days prior to sacrifice) and below, with a slope of 37.7 pmol CEVal/g globin/p.p.m. At higher concentrations, adduct levels increased sublinearly, indicating saturation of a metabolic process for elimination of ACN. Comparison of adduct formation with the estimated dose (mg/kg/day) of ACN indicated that at low dose (0-10 p.p.m.) CEVal = 0.508 x ACN dose + 0.048 and at high dose (35-300 p.p.m.) CEVal = 1.142 x ACN dose - 1.098. Globin from the smokers (10-20 cigarettes/day) contained about 90 pmol CEVal/g, whereas the adduct levels in globin from non-smokers were below the detection limit. The analytical sensitivity should be sufficient to allow monitoring of occupationally exposed workers at levels well below the current Occupational Safety and Health Administration standard of 2 p.p.m.
authors.affiliation: Chemical Industry Institute of Toxicology, Research Triangle Park, NC 27709.
authors.forename: S M
authors.initials: SM
authors.lastname: Osterman-Golkar
authors.forename: J P
authors.initials: JP
authors.lastname: MacNeela
authors.forename: M J
authors.initials: MJ
authors.lastname: Turner
authors.forename: V E
authors.initials: VE
authors.lastname: Walker
authors.forename: J A
authors.initials: JA
authors.lastname: Swenberg
authors.forename: S J
authors.initials: SJ
authors.lastname: Sumner
authors.forename: N
authors.initials: N
authors.lastname: Youtsey
authors.forename: T R
authors.initials: TR
authors.lastname: Fennell
mesh_terms: D000181:Acrylonitrile / Q000032:analysis* / Q000494:pharmacology; D000595:Amino Acid Sequence; D000818:Animals; D004781:Environmental Exposure; D004784:Environmental Monitoring; D006454:Hemoglobins / Q000737:chemistry / Q000187:drug effects*; D006801:Humans; D008969:Molecular Sequence Data; D051381:Rats; D011916:Rats, Inbred F344; D012680:Sensitivity and Specificity; D012907:Smoking; D014633:Valine / Q000031:analogs & derivatives* / Q000737:chemistry / Q000187:drug effects
chemical_list: D006454:Hemoglobins; C091192:N-(2-cyanoethyl)valine; D014633:Valine; D000181:Acrylonitrile