Studying the ability of the food additive E171 (titanium dioxide) to induce gene mutations in bacteria

Anastasiya A. Tsareva, Olga V. Egorova, Yuliya V. Demidova, N. A. Ilyushina
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Abstract

Introduction. Titanium dioxide in the Russian Federation is approved for use in the food industry, in the production of medicines and hygiene products. The food additive E171 is a mixture of micro- and nanoparticles of TiO2. In 2010, IARC classified TiO2 in nanoform as a probably carcinogenic to humans (Group 2B). In vitro and in vivo studies of the genotoxicity of titanium dioxide revealed contradictory results, indicating both the presence and absence of TiO2 mutagenicity. The aim of the work is to evaluate the mutagenicity of the food additive E171 in the Ames test using standard and modified protocols. Materials and methods. The ability of food additive E171 (China) to induce reverse gene mutations in 5 strains of Salmonella typhimurium was studied under standard and modified conditions (cultivation of bacteria in the presence of methylated b-cyclodextrin (MCD) and/or pre-incubation for 1 hour in potassium phosphate buffer, pH 5.5 containing 10 mM NaCl and/or 3M MCD). Results. A sample of food additive E171 based on rutile titanium dioxide does not induce gene mutations in S. typhimurium in standard experiments. Modification of the Ames test protocol (decrease of the incubation mixture pH, addition of 10 mM NaCl) revealed statistically significant dose-dependent effects in TA100, TA98, and TA97 strains under metabolic incubation conditions. However, the fold increase of the number of revertants in the experimental plates compared to the negative control was < 2. Limitations. The research is limited to the mutagenicity assessment of food additive E171 (titanium dioxide) in the Ames test. Conclusion. The evaluation of the mutagenicity of titanium dioxide in other in vitro and in vivo tests taking into account the size and shape of the particles, is necessary to resolve the issue of its genetic safety as a food dye. A full range of studies will be performed on other samples of titanium dioxide presented in the market of the Russian Federation.
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研究食品添加剂 E171(二氧化钛)诱导细菌基因突变的能力
简介。俄罗斯联邦批准将二氧化钛用于食品工业、药品和卫生产品的生产。食品添加剂 E171 是二氧化钛微粒和纳米粒子的混合物。2010 年,国际癌症研究机构(IARC)将纳米形态的二氧化钛列为可能对人类致癌的物质(2B 组)。对二氧化钛的遗传毒性进行的体外和体内研究显示了相互矛盾的结果,既表明二氧化钛存在致突变性,也表明二氧化钛不存在致突变性。 这项工作的目的是采用标准和修改后的方案,在艾姆斯试验中评估食品添加剂 E171 的致突变性。 材料和方法。在标准和改进条件下(在甲基化 b-环糊精(MCD)存在下培养细菌和/或在含有 10 mM NaCl 和/或 3M MCD 的 pH 值为 5.5 的磷酸二氢钾缓冲液中预培养 1 小时),研究了食品添加剂 E171(中国)诱导 5 株鼠伤寒沙门氏菌反向基因突变的能力。 结果在标准实验中,基于金红石二氧化钛的食品添加剂 E171 样品不会诱发鼠伤寒杆菌的基因突变。在新陈代谢培养条件下,修改艾姆斯试验方案(降低培养混合液的 pH 值,添加 10 mM NaCl)对 TA100、TA98 和 TA97 菌株的影响具有统计学意义的剂量依赖性。然而,与阴性对照组相比,实验平板中逆转录酶数量的增加倍数小于 2。 局限性。研究仅限于在艾姆斯试验中对食品添加剂 E171(二氧化钛)进行诱变性评估。 结论。为了解决二氧化钛作为食品染料的基因安全性问题,有必要在其他体外和体内试验中对二氧化钛的致突变性进行评估,同时考虑到颗粒的大小和形状。将对俄罗斯联邦市场上的其他二氧化钛样品进行全面研究。
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