Article DNA damage in human lymphocytes exposed to four food additives in vitro Serkan Yilmaz 1 , Fatma U ¨ nal 2 , Deniz Yu ¨ zbas ¸ıog ˘lu 2 and Mustafa C ¸ elik 3 Abstract In vitro genotoxic effects of antioxidant additives, such as citric acid (CA) and phosphoric acid (PA) and their combination, as well as antimicrobial additives, such as benzoic acid (BA) and calcium propionate (CP), on human lymphocytes were determined using alkaline single-cell gel electrophoresis. There was a significant increase in the DNA damage in human lymphocytes after 1 h of in vitro exposure to CA, PA, BA and CP (200, 25–200, 50–500, 50–1000 mg/mL, respectively). The combination of CA and PA significantly increased the mean tail intensity at all the concentrations used (25–200 mg/mL) and significantly increased the mean tail length mainly after higher concentrations (100 and 200 mg/mL). Data in this study showed that the concentra- tions of food additives used induce DNA damage and PA was the most genotoxic and CA was less genotoxic additives among them. Keywords Food additives, citric acid, phosphoric acid, benzoic acid, calcium propionate, comet assay Introduction Food additives are the substances that are added to food in order to prolong the shelf-life of the factory made foods by inhibiting the development of bacteria, fungi and other microorganisms. They are also used for some other purposes including coloring, flavoring, sweetening and thickening (Rekna and Dharman, 2011). There are about 2500 chemicals that function as food additives that give rise to about 5000 trade name products on a world wide-basis (Scotter and Castle, 2004). However, the increased consumption of food additives may cause toxic reactions. It was reported that some food additives have genotoxic and carcinogenic effects in different test organisms including bacteria, plants, human lymphocytes, mice and rats (Mamur et al., 2010; Mpountoukas et al., 2008; Sasaki et al., 2002; Yılmaz et al., 2008a, 2008b, 2009). In late 1988, the European Community adopted a directive that set out the criteria by which the addi- tives would be assessed and established the list of additives that could be used, the foods in which they could be used and their maximum intake levels (Council Directive, 1989; Mpountoukas et al. 2008). The use of preservatives is controlled by the legislation across the European Union by formulating a specific directive (Council Directive, 1995; Mpoun- toukas et al., 2008). However, some of the food additives have been prohibited from use because of their toxicity. For example, 2-(2-furyl)-3-(5-nitro-2- furyl)acrylamide was used as a food preservative in Japan before 1965 until it was banned because of carcinogenicity in experimental animals (IARC, 1983). Butter yellow (p-dimethylaminoazobenzene), an azo compound, was listed for use in food in the United States and delisted in the same year; legisla- tion prohibiting its use in Europe followed soon after, because it was implicated as a carcinogen in several animal species (IARC, 1975). Many azo compounds were found to be genotoxic in short-term tests and 1 Faculty of Health Sciences, Ankara University, Ankara, Turkey 2 Department of Biology, Science Faculty, Gazi University, Ankara, Turkey 3 Department of Medical Biology, Faculty of Medicine, Kahramanmaras ¸ Su ¨tc ¸u ¨ _ Imam University, Kahramanmaras ¸, Turkey Corresponding author: Fatma U ¨ nal, Department of Biology, Science Faculty, Gazi University, Ankara 06500, Turkey. Email: funal@gazi.edu.tr Toxicology and Industrial Health 2014, Vol. 30(10) 926–937  The Author(s) 2012 Reprints and permissions: sagepub.co.uk/journalsPermissions.nav DOI: 10.1177/0748233712466132 tih.sagepub.com at Gazi University on January 21, 2016 tih.sagepub.com Downloaded from