基于高分辨质谱技术的农药与生物大分子相互作用的分析方法  

Analysis of Interaction between Pesticide and Biological Macromolecules Based on High Resolution Mass Spectrometry

作　　者：李康丽 王玲玲[1] 暴雅静 熊博[1] 王震宇[1] 

机构地区：[1]华中师范大学农药与化学生物学教育部重点实验室,武汉430079

出　　处：《分析化学》2016年第2期218-223,共6页Chinese Journal of Analytical Chemistry

基　　金：国家自然科学基金(Nos.21205044;21302060);高校教育部基础研究经费(Nos.CCNU14A05005;CCNU15KFY004)的资助~~

摘　　要：建立了检测农药小分子与生物大分子(BSA蛋白,SOD酶)相互作用的高分辨质谱分析方法。对相应结合产物进行质谱检测,结果表明,BSA蛋白与甲基对硫磷分子在开始相互作用30 min后达到饱和,且每个蛋白分子最多与5个甲基对硫磷分子结合;BSA蛋白与甲维盐不存在相互作用。通过对SOD酶与敌敌畏,阿维菌素及噻呋酰胺结合产物的质谱检测发现每个SOD酶最多结合2个敌敌畏分子,1个阿维菌素分子,且不与噻呋酰胺相互作用。此外,实验表明,SOD酶与阿维菌素分子作用30 min后达到平衡,与敌敌畏分子作用20 min后达到平衡。通过对两种蛋白结合农药小分子过程的时间分辨分析发现,两种生物大分子结合农药小分子的机理过程存在差异。本方法与相关标准方法(荧光光谱法,NBT试剂盒法)及分子模拟对接结果比对说明,本方法切实可靠。本方法具有耗样量少、检测速度快、可提供多方面信息等优点,在新农药的研发及其安全性评价方面具有实用价值。An analytical method for the determination of maeromolecules （bovine serum albumin （BSA） protein, developed based on high resolution mass spectrometry. All interaction between pesticide superoxide dismutase （ SOD ） and biological enzyme ） was results revealed that each BSA bonded with maximum five Parathion-methyls after 30 min, and it would not interact with emamectin benzoate. Moreover, each SOD after mixing with corresponding pesticides could bond with two dichlorvos （DDVPs） and one avermectin after 30 min. In addition, there is no interaction between SOD and thifluzamide. The interaction between SOD and avermectin trended stable after 30 min, and similar phenomenon was observed between SOD and DDVP after 20 min. There were certain obvious some differences in the pathway of interactions for BSA and SOD, both of which were bonded with the pesticides, based on time resolved determinations for their bonding process. Finally, we compared the proposed method with standard methods （fluorescence spectrometry, NBT kit method） and molecular docking results, and the reliability of the proposed method was supported. It owned preponderances in less sample consumption, high detection speed, and more capacities compared with traditional strategies. Therefore, the proposed mothed would own certain practical values in either researches and developments or safety evaluations for new pesticides.

关 键 词：高分辨质谱 分子相互作用 农药 牛血清白蛋白 超氧化物歧化酶 

分 类 号：O657.63[理学—分析化学] TQ450.1[理学—化学]