Genotoxicity, mutagenicity and cytotoxicity of carotenoids extracted from ionic liquid in multiples organs of Wistar rats Paula Martins Larangeira, Veridiana Vera de Rosso, Victor Hugo Pereira da Silva, Carolina Foot Gomes de Moura, Daniel Araki Ribeiro, DDS, PhD* Department of Biosciences, Federal University of Sao Paulo, UNIFESP, SP, Brazil A R T I C L E I N F O Article history: Received 10 June 2016 Received in revised form 17 August 2016 Accepted 7 September 2016 Keywords: Ionic liquid Lycopene Mutagenicity Genotoxicity Rat A B S T R A C T The ionic liquid or melted salt 1-Butyl-3-methylimidazolium is an alternative process to extract natural pigments, such as carotenoids. Lycopene represents 80–90% of total of carotenoids presents in tomatoes and it has been widely studied due its potent antioxidant action. The aim of this study was to evaluate genotoxicity, mutagenicity and cytotoxicity of carotenoids extracted from ionic liquid using experimental model in vivo. For this purpose, a total of 20 male Wistar rats were distributed into four groups (n = 5), as follows: control group; received a corresponding amount of corn oil for 7 days by intragastric gavage (i.g.), ionic liquid group, received 10 mg kg 1 body weight for 7 days by gavage; 10 mg carotenoids group, received 10 mg kg 1 bw dissolved in corn oil for 7 days by gavage and 500 mg carotenoids group, received 500 mg kg 1 bw dissolved in corn oil for 7 days by gavage. Rat liver treated with ionic liquid exhibited moderate histopathological changes randomly distributed in the parenchyma, such as cytoplasmic eosinophilia, apoptotic bodies, inflammatory infiltrate and focal necrosis. DNA damage was found in peripheral blood and liver cells of rats treated with ionic liquid or carotenoids at 500 mg. An increase of micronucleated cells and 8-OhDG immunopositive cells were also detected in rats treated with carotenoids at 500 mg. In summary, our results demonstrate that recommended dose for human daily intake of carotenoids extracted by ionic liquid did not induce genotoxicity, mutagenicity and cytotoxicity in multiple organs of rats. ã 2016 Elsevier GmbH. All rights reserved. 1. Introduction Ionic liquid is molten salts formed by an organic cation and different chemical class of anion. Its ionic properties have been used as extractor solvent under different applications and paradigms (Bi et al., 2010). Therefore, the use of ionic liquid is environmentally acceptable and it is recommended as sustainable processes of extraction, generating profitability and recognition among several companies dedicated to this purpose. An important advantage of ionic liquid resides on the fact that its ionic characteristics are high density and low vapor pressure facilitating the isolation process of organic compounds, which are usually slightly soluble (Dupont et al., 2002). Carotenoids are good examples of compounds obtained from chemical synthesis and they are employed in the food industry. The conventional method for extracting carotenoids is quite complicated from natural sources due to low solubility. To overcome the problem, it is used volatile organic solvents, such as acetone and hexane. These solvents cannot be used in food products since they are toxic to humans. In this context, the development of new extraction techniques employing ionic liquids becomes relevant and promising so far. Among carotenoids, lycopene has high coloring strength and chemical/biological activities against reactive oxygen species (ROS). Numerous studies have proved that lycopene exerts beneficial effect on human health (Boyacioglu et al., 2016). Lycopene contains approximately 80–90% of total carotenoids present in tomatoes being widely distributed throughout fruit (Del Valle et al., 2006). Genotoxicity, mutagenicity and cytotoxicity studies may be used as biomarkers for under- standing the adverse effects of ionic liquid and carotenoids after ingestion (Jodynis-Liebert et al., 2009; Aydin et al., 2013). Histopathological analysis, DNA damage, DNA repair capacity, and micronucleus are potential biomarkers traditionally used for this purpose. As a result and because of the lack of scientific evidence, the aim of the present study was to evaluate * Corresponding author at: Departamento de Biociências, Universidade Federal de São Paulo – UNIFESP, Av. Ana Costa 95, Zip code: 11060-001, Santos – SP, Brazil. E-mail addresses: daribeiro@unifesp.br , ak92@hotmail.com (D.A. Ribeiro). http://dx.doi.org/10.1016/j.etp.2016.09.003 0940-2993/ã 2016 Elsevier GmbH. All rights reserved. Experimental and Toxicologic Pathology xxx (2016) xxx–xxx G Model ETP 50976 No. of Pages 8 Please cite this article in press as: P.M. Larangeira, et al., Genotoxicity, mutagenicity and cytotoxicity of carotenoids extracted from ionic liquid in multiples organs of Wistar rats, Exp Toxicol Pathol (2016), http://dx.doi.org/10.1016/j.etp.2016.09.003 Contents lists available at ScienceDirect Experimental and Toxicologic Pathology journal homepage: www.else vie r.de /et p