ABSTRACT
Allergic and irritant skin reactions caused by topical exposure to permanent hair dyes are a common problem. For regulatory and ethnical purposes, it is required to perform chemical safety assessment following the replacement, reduction, and refinement of animal testing (3Rs). Permanent hair dyes are formed by a mixture of ingredients that vary from low to extreme skin sensitizing potency and that inter-react to form unknown by-products. Because of the complex reaction, this cytotoxic mechanism has not yet been elucidated and is the subject of this study. Here, we topically exposed p-phenylenediamine (PPD), Resorcinol (RES), Hydrogen Peroxide (H2O2) alone or as a mixture to RhE and evaluated parameters related to skin irritation such as epidermal viability, keratinocytes damage, barrier loss and IL-1 α. Our data indicates that ingredients tested alone did not lead to an increase of cytotoxic parameters related to skin irritation. However, when the mixture of PPD/H2O2/RES and PPD/H2O2 was applied to the RhE, some of the parameters such as morphological changes including the presence of apoptotic cells, barrier loss and increased IL- 1 α release were observed. The results indicate that the mixture of ingredients used in permanent hair dyes have an irritant effect in RhE while the ingredients alone not.
Subject(s)
Allergens/toxicity , Epidermis/drug effects , Hair Dyes/toxicity , Interleukin-1alpha/immunology , Skin/drug effects , Cells, Cultured , Epidermis/immunology , Epidermis/injuries , Humans , Interleukin-1alpha/genetics , Keratinocytes/drug effects , Keratinocytes/immunology , Skin/immunologyABSTRACT
This work describes the mutagenic response of Sudan III, an adulterant food dye, using Salmonella typhimurium assay and the generation of hazardous aromatic amines after different oxidation methods of this azo dye. For that, we used metabolic activation by S9, catalytic oxidation by ironporphyrin and electrochemistry oxidation in order to simulate endogenous oxidation conditions. The oxidation reactions promoted discoloration from 65% to 95% of Sudan III at 1 × 10(-4)molL(-1) and generation of 7.6 × 10(-7)molL(-1) to 0.31 × 10(-4)molL(-1) of aniline, o-anisidine, 2-methoxi-5-methylaniline, 4-aminobiphenyl, 4,4'-oxydianiline; 4,4'-diaminodiphenylmethane and 2,6-dimethylaniline. The results were confirmed by LC-MS-MS experiments. We also correlate the mutagenic effects of Sudan III using S. typhimurium with the strain TA1535 in the presence of exogenous metabolic activation (S9) with the metabolization products of this compound. Our findings clearly indicate that aromatic amines are formed due to oxidative reactions that can be promoted by hepatic cells, after the ingestion of Sudan III. Considering that, the use of azo compounds as food dyestuffs should be carefully controlled.
Subject(s)
Amines/metabolism , Azo Compounds/metabolism , Azo Compounds/toxicity , Coloring Agents/metabolism , Cytochrome P-450 Enzyme System/metabolism , Amines/analysis , Amines/toxicity , Animals , Azo Compounds/chemistry , Catalysis , Coloring Agents/chemistry , Coloring Agents/toxicity , Electrochemical Techniques , Food Coloring Agents/toxicity , Isoenzymes/metabolism , Liver/enzymology , Metalloporphyrins/chemistry , Metalloporphyrins/metabolism , Mutagenicity Tests/methods , Mutation , Oxidation-Reduction , Rats , Rats, Sprague-Dawley , Salmonella typhimurium/drug effects , Salmonella typhimurium/genetics , Tandem Mass SpectrometryABSTRACT
Indigo carmine forms a stable complex with different ions, and the stability constant of the complexes were evaluated as log K equal to 5.75; 5.00; 4.89 and 3.89 for complexes with Cu(II), Ni(II), Co(II) and Zn(II) ions, respectively, in 0.1 mol L-1 carbonate buffer solution at pH 10. The interaction between Cu(II) ions and indigo carmine (IC) in alkaline medium resulted in the formation of the Cu2(IC) complex, measured by the spectrophotometric method, with a stoichiometric ratio between indigo carmine and metal ions of 2:1 (metal-ligand). The reported method has also been successfully tested for determination of copper in pharmaceutical compounds based on copper-gluconate without pre-treatment.
Índigo carmim forma complexos estáveis com diferentes íons e a constante de estabilidade dos complexos foi avaliada como log K igual 5,75; 5,00; 4,89 e 3,89, respectivamente, para os complexos com os íons Cu(II), Ni(II), Co(II) e Zn(II) em solução tampão carbonato 0,1 mol L-1, pH 10. A interação entre o íon Cu(II) e índigo carmin (IC) em meio alcalino resultou na formação do complexo Cu2(IC) monitorado por método espectrofotométrico, com razão estequiométrica entre o índigo carmim e o íon metálico de 2:1 (metal-ligante). O método relatado também tem sido testado com sucesso para determinação de cobre em compostos farmacêuticos à base de cobre-gliconato sem qualquer pré-tratamento.
