Please cite this article in press as: F.M.D. Chequer, et al., The cosmetic dye quinoline yellow causes DNA damage in vitro, Mutat. Res.: Genet. Toxicol. Environ. Mutagen. (2014), http://dx.doi.org/10.1016/j.mrgentox.2014.11.003 ARTICLE IN PRESS G Model MUTGEN 402555 1–8 Mutation Research xxx (2014) xxx–xxx Contents lists available at ScienceDirect Mutation Research/Genetic Toxicology and Environmental Mutagenesis jo ur nal home page: www.elsevier.com/locate/gentox Comm unit y ad dress: www.elsevier.com/locate/mutres The cosmetic dye quinoline yellow causes DNA damage in vitro Farah Maria Drumond Chequer a,b,∗ , Vinícius de Paula Venâncio a , Q1 Maíra Rocha de Souza Prado a , Luiz Raimundo Campos da Silva e Cunha Junior c , Thiago Mescoloto Lizier d , Maria Valnice Boldrin Zanoni d , Rommel Rodríguez Burbano c , Maria Lourdes Pires Bianchi a , Lusânia Maria Greggi Antunes a a Departamento de Análises Clínicas, Toxicológicas e Bromatológicas, Faculdade de Ciências Farmacêuticas de Ribeirão Preto, Universidade de São Paulo, USP, Ribeirão Preto, SP 14040-903, Brazil b Departamento de Análises Clínicas e Toxicológicas, Faculdade Federal de Minas Gerais, UFMG, Belo Horizonte, MG 31270-901, Brazil c Laboratório de Citogenética Humana, Instituto de Ciências Biológicas, Universidade Federal do Pará, Belém, PA, Brazil d Instituto de Química. Departamento de Química Analítica, Universidade Estadual Paulista – UNESP, Quitandinha 14800-900, Araraquara/SP, Brazil a r t i c l e i n f o Article history: Received 29 June 2014 Received in revised form 6 November 2014 Accepted 11 November 2014 Available online xxx Keywords: Genotoxicity Cosmetic dye Micronucleus Comet assay Oxidation a b s t r a c t Quinoline yellow (QY) is a chinophthalon derivative used in cosmetic compositions for application to the skin, lips, and/or body surface. However, regulatory data about the genotoxicity and/or mutageni- city of this compound are still controversial. Therefore, this work evaluated the genotoxicity of QY using the comet assay and the cytokinesis-block micronucleus cytome assay (CBMN-Cyt) in the metabolically competent cell line HepG2, which closely mimics phase I metabolism. This research also identified the products formed after electrochemical oxidation of the QY dye, which simulates hepatic biotransforma- tion. The primary products generated after the oxidation process were analyzed by High Performance Liquid Chromatography coupled with a Diode Array Detector (HPLC/DAD), which detected the production of 4,4 ′ -diaminodiphenylmethane, 2-methoxy-5-methylaniline and 4,4 ′ -oxydianiline. The results demon- strated that low (from 0.5 to 20 g mL -1 ) QY concentrations were genotoxic in HepG2 cells on both assays and those harmful compounds were detected after the oxidation process. Our findings suggest that this colorant could cause harmful effects to humans if it is metabolized or absorbed through the skin. © 2014 Published by Elsevier B.V. 1. Introduction Synthetic dyes are used extensively in many industries, including the cosmetics, textile, pharmaceutical, food, plastics, photography and paper industries [1–4]. It is estimated that over 10,000 different dyes and pigments are used industrially and that over 7 × 10 5 tons of synthetic dyes are produced annually world- wide [4–6]. However, there is insufficient information about their potential health risks for humans and the environment [7,8]. The available toxicological data about cosmetics dyes have shown effects that range from contact allergies to different types of genetic damages, including genotoxicity, mutagenicity and early age leukemia [9–13]. ∗ Corresponding author at: Departamento de Análises Clínicas, Toxicológicas e Q2 Bromatológicas, Faculdade de Ciências Farmacêuticas de Ribeirão Preto, Universi- dade de São Paulo, USP, Ribeirão Preto, SP 14040-903, Brazil. Tel.: +55 16 3602 4186; fax: +55 16 3602 4725. E-mail address: farahchequer@gmail.com (F.M.D. Chequer). The dye quinoline yellow (QY) is a chinophthalon derivative used in cosmetic compositions for application to the skin, lips, and/or body surface [14]. This dye (also known as D&C Yellow no. 11) was found to induce allergic contact dermatitis; in a human maximization test, 15 of 20 subjects became sensitized [15,16]. The regulatory data regarding QY genotoxicity and/or mutagenicity are still controversial [14,17]. Therefore, we studied the dye QY in this research. The aim of this investigation was to evaluate the genotox- icity of QY using the alkaline comet assay and the cytokinesis-block micronucleus cytome assay (CBMN-Cyt) in the metabolically com- petent cell line HepG2, which closely mimics phase I metabolism. Micronuclei (MN) were also analyzed using the fluorescence in situ hybridization (FISH) technique for further hazard characterization. In addition, it is known that aromatic amines can be produced during oxidative and/or reductive processes [3,18], and the forma- tion of these aromatic amine byproducts could be important for understanding the chemical transformation of dyes. Therefore, the present work also aimed to determine if 16 aromatic amines used as standard models of amines classified by IARC [19] could be pro- duced during the oxidative and/or reductive conditions. 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