Journal of Ethnopharmacology 135 (2011) 730–736 Contents lists available at ScienceDirect Journal of Ethnopharmacology journal homepage: www.elsevier.com/locate/jethpharm Acute, subacute toxicity and mutagenic effects of anacardic acids from cashew (Anacardium occidentale Linn.) in mice Ana Laura Nicoletti Carvalho a,∗ , Raquel Annoni a , Paula Regina Pereira Silva a , Primavera Borelli b , Ricardo Ambrósio Fock b , Maria Teresa Salles Trevisan c , Thais Mauad a,d a Experimental Atmospheric Pollution Laboratory (LPAE), Department of Pathology, São Paulo Medical School, University of São Paulo, Av Dr Arnaldo, 455, Room 1155, 01246-903 São Paulo, SP, Brazil b Experimental Hematology Laboratory, Department of Clinical and Toxicological Analyses, Faculty of Pharmaceutical Sciences, University of São Paulo, Av Prof Lineu Prestes, 580/17, 05508-900 São Paulo, SP, Brazil c Department of Organic and Inorganic Chemistry, Federal University of Ceará, CP 12200, 60451-970 Fortaleza, CE, Brazil d National Institute for Integrated Analysis of Environmental Risk (INAIRA), National Council for Scientific and Technological Development (CNPq), Ministry of Science and Technology, for the Development of National Institutes of Science and Technology (MCT), Av Dr Arnaldo, 455, Room 1220, 01246-903 São Paulo, SP, Brazil article info Article history: Received 13 December 2010 Received in revised form 9 March 2011 Accepted 3 April 2011 Available online 12 April 2011 Keywords: Anacardic acids Anacardium occidentale Acute toxicity Subacute toxicity Mutagenicity abstract Aim of the study: Anacardium occidentale Linn. (cashew) is a Brazilian plant that is usually consumed in natura and is used in folk medicine. Anacardic acids (AAs) in the cashew nut shell liquid are biologically active as gastroprotectors, inhibitors of the activity of various deleterious enzymes, antitumor agents and antioxidants. Yet, there are no reports of toxicity testing to guarantee their use in vivo models. Materials and methods: We evaluated AAs biosafety by measuring the acute, subacute and mutagenic effects of AAs administration in BALB/c mice. In acute tests, BALB/c mice received a single oral dose of 2000 mg/kg, whereas animals in subacute tests received 300, 600 and 1000 mg/kg for 30 days. Hemato- logical, biochemical and histological analyses were performed in all animals. Mutagenicity was measured with the acute micronucleus test 24 h after oral administration of 250 mg/kg AAs. Results: Our results showed that the AAs acute minimum lethal dose in BALB/c mice is higher than 2000 mg/kg since this concentration did not produce any symptoms. In subacute tests, females which received the highest doses (600 or 1000 mg/kg) were more susceptible, which was seen by slightly decreased hematocrit and hemoglobin levels coupled with a moderate increase in urea. Anacardic acids did not produce any mutagenic effects. Conclusions: The data indicate that doses less than 300 mg/kg did not produce biochemical and hematolog- ical alterations in BALB/c mice. Additional studies must be conducted to investigate the pharmacological potential of this natural substance in order to ensure their safe use in vivo. © 2011 Elsevier Ireland Ltd. All rights reserved. 1. Introduction Cashew, Anacardium occidentale Linn. is a tropical tree native to northeast Brazil. Cashew apples (pseudofruit) and nuts can be con- sumed in natura and converted into various nutritional products (juice, tea, jam and beverages), (Kubo et al., 1993; Trevisan et al., Abbreviations: AAs, anacardic acids; ALP, alkaline phosphatase; ALT, alanine aminotransferase; AST, aspartate aminotransferase; b.w., body weight; CAJ, cashew apple juice; CNU, cashew nut oil; CNSL, cashew nut shell liquid; ECPs, polychro- matic erythrocytes; MNPCEs, micronucleated polychromatic erythrocytes; MNU, N-methyl-N-nitrosourea; ROS, reactive oxygen species. ∗ Corresponding author. Tel.: +55 11 30617173; fax: +55 11 30628098. E-mail addresses: analauranc@yahoo.com.br (A.L.N. Carvalho), rqannoni@yahoo.com.br (R. Annoni), preginahpv@usp.br (P.R.P. Silva), borelli@usp.br (P. Borelli), hemato@usp.br (R.A. Fock), trevisan@ufc.br (M.T.S. Trevisan), tmauad@usp.br (T. Mauad). 2006). Cashew nut shell liquid (CNSL) is used in industrial applica- tions such as food preservatives, paints, cements and for gasoline stabilization; as such, it is an important commercial product in sev- eral tropical countries (Paramashivappa et al., 2001; Trevisan et al., 2006; Narasimhan et al., 2008). In addition, this plant has been widely used in folk medicine in Brazil, India and Africa to treat inflammation, gastrointestinal dis- eases and hypertension (Mota et al., 1985; Cavalcante et al., 2003; Konan and Bacchi, 2007b). Several studies have evaluated the bio- logical effects and pharmaceutical potential of cashew tree extracts and parts. For instance, pre-treatment with 200 mg/kg of the methanol extract of Anacardium occidentale stem bark completely protected against lipopolysaccharide-induced septic shock in Swiss mice (Olajide et al., 2004). Hydroethanolic extract from cashew leaves, which are rich in polyphenols, inhibited gastric lesions induced by HCl/ethanol in female rats (Konan and Bacchi, 2007b). Finally, a mixture of condensed and hydrolysable tannins from 0378-8741/$ – see front matter © 2011 Elsevier Ireland Ltd. All rights reserved. doi:10.1016/j.jep.2011.04.002