title: Further investigations into the genotoxicity of 2,6-xylidine and one of its key metabolites.
abstract: The metabolite of several amide anaesthetics, 2,6-xylidine, is a possible human (Group 2B) carcinogen and induced nasal tumours in rats after dietary administration. However, published papers on the genotoxicity of 2,6-xylidine in vitro have given inconsistent results. It has been proposed that the genotoxicity of 2,6-xylidine is dependent on its metabolism to a key metabolite dimethylphenyl N-hydroxylamine (DMHA), which would then be further converted to form a reactive nitrenium ion by phase 2 (mainly acetylation) metabolism. In order to study whether the inconsistent results could be explained by different systems having different potential for DMHA to be formed and to induce genotoxicity in vitro, we have tested 2,6-xylidine in conventional Ames bacteria, and strains engineered to overexpress acetyltransferase, in the presence of different concentrations of induced rat liver and human liver S9. All tests gave consistently negative results. The formation of DMHA by induced rat liver S9 and human S9 was clearly shown to occur, and to be concentration- and time-dependent. The potential inhibitory effects of the solvent DMSO were also studied, but it was clearly not responsible for the negative results with 2,6-xylidine. Thus, whatever is the mode of action of 2,6-xylidine carcinogenicity in rodents, it has proven impossible to detect mutagenic effects in Ames tests with numerous variations of metabolic conditions, or even using acetyltransferase overexpressing strains of bacteria.
authors.affiliation: Kirkland Consulting, P.O. Box 79, Tadcaster LS24 0AS, United Kingdom. dkirkland@genetoxconsulting.co.uk
authors.forename: David
authors.initials: D
authors.lastname: Kirkland
authors.forename: Mark
authors.initials: M
authors.lastname: Ballantyne
authors.forename: Stefanie
authors.initials: S
authors.lastname: Harlfinger
authors.forename: Olaf
authors.initials: O
authors.lastname: Will
authors.forename: Ulrich
authors.initials: U
authors.lastname: Jahnel
authors.forename: Alexander
authors.initials: A
authors.lastname: Kraus
authors.forename: Corinne van
authors.initials: Cv
authors.lastname: Dorp
mesh_terms: D000814:Aniline Compounds / Q000378:metabolism / Q000633:toxicity*; D000818:Animals; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism; D004926:Escherichia coli / Q000187:drug effects / Q000235:genetics; D006801:Humans; D006898:Hydroxylamines / Q000378:metabolism / Q000633:toxicity*; D008099:Liver / Q000378:metabolism; D009152:Mutagenicity Tests; D051381:Rats; D011994:Recombinant Proteins / Q000378:metabolism; D012486:Salmonella typhimurium / Q000187:drug effects / Q000235:genetics
chemical_list: D000814:Aniline Compounds; D006898:Hydroxylamines; D011994:Recombinant Proteins; C007766:2,6-xylidine; D003577:Cytochrome P-450 Enzyme System; C036684:2,4-dimethylphenylhydroxylamine