title: N-acetylation of three aromatic amine hair dye precursor molecules eliminates their genotoxic potential.
abstract: N-acetylation has been described as a detoxification reaction for aromatic amines; however, there is only limited data available showing that this metabolic conversion step changes their genotoxicity potential. To extend this database, three aromatic amines, all widely used as precursors in oxidative hair dye formulations, were chosen for this study: p-phenylenediamine (PPD), 2,5-diaminotoluene (DAT) and 4-amino-2-hydroxytoluene (AHT). Aiming at a deeper mechanistic understanding of the interplay between activation and detoxification for this chemical class, we compared the genotoxicity profiles of the parent compounds with those of their N-acetylated metabolites. While PPD, DAT and AHT all show genotoxic potential in vitro, their N-acetylated metabolites completely lack genotoxic potential as shown in the Salmonella typhimurium reversion assay, micronucleus test with cultured human lymphocytes (AHT), chromosome aberration assay with V79 cells (DAT) and Comet assay performed with V79 cells. For the bifunctional aromatic amines studied (PPD and DAT), monoacetylation was sufficient to completely abolish their genotoxic potential. Detoxification through N-acetylation was further confirmed by comparing PPD, DAT and AHT in the Comet assay using standard V79 cells (N-acetyltransferase (NAT) deficient) and two NAT-proficient cell lines,V79NAT1*4 and HaCaT (human keratinocytes). Here we observed a clear shift of dose-response curves towards decreased genotoxicity of the parent aromatic amines in the NAT-proficient cells. These findings suggest that genotoxic effects will only be found at concentrations where the N-acetylation (detoxifying) capacity of the cells is overwhelmed, indicating that a 'first-pass' effect in skin could be taken into account for risk assessment of these topically applied aromatic amines. The findings also indicate that the use of liver S-9 preparations, which generally underestimate Phase II reactions, contributes to the generation of irrelevant positive results in standard genotoxicity tests for this chemical class.
authors.affiliation: The Procter and Gamble Co., Cosmital SA, Route de Chésalles 21, 1723 Marly, Switzerland.
authors.forename: Andreas
authors.initials: A
authors.lastname: Zeller
authors.forename: Stefan
authors.initials: S
authors.lastname: Pfuhler
mesh_terms: D000107:Acetylation; D000588:Amines / Q000737:chemistry / Q000378:metabolism / Q000633:toxicity*; D000814:Aniline Compounds / Q000633:toxicity; D000818:Animals; D002461:Cell Line, Transformed; D002869:Chromosome Aberrations / Q000187:drug effects; D020552:Comet Assay; D003408:Cresols / Q000633:toxicity; D005260:Female; D006202:Hair Dyes / Q000737:chemistry / Q000378:metabolism / Q000633:toxicity*; D006801:Humans; D008297:Male; D015162:Micronucleus Tests; D015394:Molecular Structure; D009152:Mutagenicity Tests; D010655:Phenylenediamines / Q000633:toxicity; D051381:Rats; D012486:Salmonella typhimurium / Q000187:drug effects
chemical_list: D000588:Amines; D000814:Aniline Compounds; D003408:Cresols; D006202:Hair Dyes; D010655:Phenylenediamines; C007303:2,5-diaminotoluene; C029728:4-phenylenediamine; C538896:4-amino-2-hydroxytoluene