基于化学计量学的合成食品添加剂体外DNA靶向作用检测技术及其作用机制研究

滥用化学合成食品添加剂是危害食品安全的重要因素之一。研究表明，过量使用合成食品添加剂会引起急性或慢性毒性甚至诱发癌变，且此诱变性与食品添加剂作用于DNA密切相关。然而目前针对二者相互作用机制的研究还非常缺乏，相互作用的检测技术也严重滞后。本项目拟以小牛胸腺DNA为靶点，构建化学计量学模型多元曲线分辨-交替最小二乘法与荧光和紫外光谱法联用新技术，应用于检测BHA等合成食品添加剂与DNA的相互作用；运用圆二色谱、红外光谱和分子模拟等手段，预测、验证并阐明合成食品添加剂的体外DNA靶向作用模式、作用位点及其损伤DNA的分子机制，探明稳定复合物的结构特征，测定相互作用的结合参数；探索合成食品添加剂的诱变性与其作用DNA的位点和引起DNA损伤之间的相互关系，揭示其可能的规律性。本项目的实施对于深入认识潜在诱变性合成食品添加剂的毒性机理和探寻新颖、有效的DNA靶向作用检测技术具有重要的理论和实践意义。

The abuse of the chemical synthesis food additives is one of the important risk factors to food safety. Studies show that the excessive use of the synthetic food additives can cause acute or chronic toxicity and even induce cancer, and the mutagenicity of the food additives is closely related to their DNA targeted interaction. However, not only the studies on the interaction mechanism between the synthetic food additives and DNA are very lack, but also the detection methods of their interaction are also serious lag. This project intends to construct a new technique, which uses calf thymus DNA as a target and adopts the strategy combined the multivariate curve resolution-alternating least squares chemometrics algorithm with fluorescence and UV-vis absorption spectroscopic methods. This technique can be applied to determine the interacton of the chemical synthesis food additives with DNA. Predict,confirm and elucidate the binding modes and binding sites of the synthetic food additives with DNA as well as the molecule mechanism of damage DNA with the use of various methods including circular dichroism, Fourier transform infrared spectroscopy, molecular modeling and so on. Clarify the structure characteristics of the stabilized DNA-food additive complexes, and determine the binding parameters between the synthetic food additives and DNA. Moreover,the correlation and regularity between the mutagenicity of the synthetic food additives and the binding sites on DNA and the structural damage of DNA will be explored. The implementation of this project has important significance for further understanding toxicity mechanism of the synthetic food additives with potential mutagenicity along with exploring innovative and effective detection technique for the interaction of DNA with the synthetic food additives.