title: In vitro genotoxicity testing of (1-chloroethenyl)oxirane, a metabolite of beta-chloroprene.
abstract: (1-Chloroethenyl)oxirane (CEO) is a metabolite of beta-chloroprene (2-chloro-1,3-butadiene, CD). The purpose of this study was to evaluate the in vitro mutagenic and clastogenic (chromosome breaking) potential of CEO. For comparative purposes, the study also included an evaluation of the racemic compounds, 3,4-epoxy-1-butene (EB) and 1,2:3,4-diepoxybutane (DEB). Mutagenicity was evaluated in a bacterial reverse mutation test (Ames), using the pre-incubation method in the presence and absence of an exogenous metabolism system (Aroclor)-induced rat liver S9). Four Salmonella typhimurium tester strains, TA97a, TA98, TA100 and TA1535 were used. The exposure concentrations in the sealed incubation vials ranged from 0 to 69 mM for CEO, 0 to 102 mM for EB, and 0 to 83 mM for DEB. All three compounds showed signs of toxicity, with DEB being substantially more toxic than either CEO or EB. Mutagenic activity was observed with all three chemicals in primarily the base pair substitution strains (S. typhimurium TA100 and TA1535), but some activity was also seen in the frameshift elimination strains (S. typhimurium TA97a and TA98). The observed mutagenic responses after exposure with CEO or EB were greater than the observed response for DEB, most likely because of the higher toxicity of DEB. Generally, the mutagenic responses were unchanged in the frameshift strains and base pair substitution strains in the presence of S9 metabolism. In vitro clastogenicity was evaluated using the cytochalasin-B blocked micronucleus test in cultured Chinese hamster V79 cells. The test was conducted without S9 metabolism because of the absence of substantial changes in the Ames test. Exposure concentrations ranged from 0 to 0.943 mM for CEO, 0 to 3.0 mM for EB, and 0 to 0.035 mM for DEB, with the upper exposure concentrations dictated by cytotoxicity. Cytotoxicity, measured as a reduction in the proportion of binucleated cells and altered cell morphology, was observed for CEO at concentrations > or =0.175 mM. Exposure to EB led to a reduced proportion of binucleated cells at concentrations > or =2.0 mM, and cell death was observed after DEB exposure at concentrations > or =0.025 mM. No clastogenicity was observed in the V79 cells when tested up to cytotoxic concentrations of CEO, whereas an elevated frequency of micronuclei was observed after exposure to either EB (> or =1.0 mM) or DEB (> or =0.0125 mM). These results suggest that CEO-induced mutagenicity, but not clastogenicity, may contribute to the observed beta-chloroprene-induced carcinogenicity in the rodent bioassay studies.
authors.affiliation: E.I. du Pont de Nemours and Company, Haskell Laboratory for Toxicology and Industrial Medicine, PO Box 50, 1090 Elkton Road, Newark, DE 19711, USA. matthew.w.himmelstein@usa.dupont.com
authors.forename: M W
authors.initials: MW
authors.lastname: Himmelstein
authors.forename: N L
authors.initials: NL
authors.lastname: Gladnick
authors.forename: E M
authors.initials: EM
authors.lastname: Donner
authors.forename: R D
authors.initials: RD
authors.lastname: Snyder
authors.forename: R
authors.initials: R
authors.lastname: Valentine
mesh_terms: D000818:Animals; D002737:Chloroprene / Q000378:metabolism* / Q000633:toxicity*; D004852:Epoxy Compounds / Q000378:metabolism / Q000633:toxicity; D005027:Ethylene Oxide / Q000031:analogs & derivatives / Q000378:metabolism* / Q000633:toxicity*; D016368:Frameshift Mutation; D066298:In Vitro Techniques; D008099:Liver / Q000378:metabolism; D009152:Mutagenicity Tests; D009153:Mutagens / Q000378:metabolism* / Q000633:toxicity*; D017354:Point Mutation; D051381:Rats; D012486:Salmonella typhimurium / Q000187:drug effects / Q000235:genetics
chemical_list: C439367:(1-chloroethenyl)oxirane; D004852:Epoxy Compounds; D009153:Mutagens; D002737:Chloroprene; C024370:3,4-epoxy-1-butene; C007366:diepoxybutane; D005027:Ethylene Oxide