Journal of Chromatography B ( IF 3 ) Pub Date : 2020-12-17 , DOI: 10.1016/j.jchromb.2020.122503 Shaogang Chu , Robert J. Letcher
The present study developed an analytical technique to investigate the possible covalent adduct formation of albumin with the herbicide atrazine, and to characterize the protein modifications in vitro using liquid chromatography separation coupled with high resolution time-of-flight mass spectrometry (LC-TOF-MS). Tandem mass spectrum analysis (MS/MS) with collision induced dissociation (CID) revealed the specific sites of rat, human and bovine serum albumin adduct with atrazine. The formation of b-ion, y-ion series in MS/MS showed a covalent adduct with an addition mass of 179.1 Da located on Cys-34 of serum albumin from rats, human and bovine. This clearly indicated that the chemical group C8H13N5 forms an adduct with Cys-34 despite the sequences differences between of rat, human and bovine serum albumin. To confirm the method reliability, concentration-dependent and time-dependent formation of adducts between serum albumins and atrazine were also investigated. Our results confirmed that atrazine can directly react with Cys-34 of serum albumin and form covalent adducts without prior metabolism.
中文翻译:
at去津形成的血清白蛋白共价加合物的液相色谱质谱鉴定与表征
本研究开发了一种分析技术,以研究白蛋白与除草剂at去津可能形成的共价加合物,并使用液相色谱分离与高分辨率飞行时间质谱(LC-TOF-MS)体外表征蛋白质修饰)。带有碰撞诱导解离(CID)的串联质谱分析(MS / MS)显示了大鼠,人和牛血清白蛋白与阿特拉津的加成反应的特定位点。MS / MS中b离子,y离子系列的形成显示出共价加合物,其附加质量为179.1 Da,位于大鼠,人和牛的血清白蛋白Cys-34上。这清楚地表明化学基团C 8 H 13 N 5尽管大鼠,人和牛血清白蛋白之间存在序列差异,但仍与Cys-34形成加合物。为了证实该方法的可靠性,还研究了血清白蛋白与at去津之间加合物的浓度依赖性和时间依赖性。我们的结果证实,阿特拉津可以直接与血清白蛋白的Cys-34反应并形成共价加合物,而无需事先代谢。