采用高效液相色谱与电感耦合等离子体质谱联用技术,对不同形态汞暴露后的Medaka肝脏和脑组织中水溶性蛋白进行研究,以体积排阻柱为液相分离柱,流动相为0.15 mol/L NH4Ac,流速为0.5 mL/min。利用电感耦合等离子体质谱对Medaka肝脏和脑组织提取液中铜、锌和汞的含量进行了分析,检出限为0.01~0.05μg/L,元素的加标回收率为93.1%~97.0%。通过与空白组对比发现,汞暴露后的Medaka鱼体肝脏和脑组织对水体中的汞有一定程度的富集;不同形态汞暴露后,在Medaka肝脏和脑组织中的分布明显不同,并且都会在一定程度上降低Medaka肝脏和脑组织中铜与锌的含量,有利于进一步研究汞在生物体内的毒性机理。
The hydrophilic mercury-binding proteins in the brain and liver extractions of Medaka after methylmercury and inorganic mercury exposure were analyzed by high performance liquid chromato-graphy coupled with inductively coupled plasma mass spectrometry.Size exclusion chromatography was selected in the experiment with a mobile phase of 0.15 mol/L NH4Ac,running at a 0.5 mL/min flow rate.Concentrations of total copper,zinc and mercury in the brain and liver extracts of Medaka were determined by ICP-MS.Detection limits for these three elements were in the range of 0.01 μg/L to 0.05 μg/L,and the spike recoveries ranged from 93.1%-97.0%.Compared with the control group,high level of mercury is accumulated in the brain and liver of exposure groups.The distribution patterns of mercury-containing proteins in the brain and liver extracts of Medaka between methylmercury and inorganic mercury exposure group were different.The concentrations of copper and zinc in the brain and liver extractions of both exposure groups were decreased.The studies of the mercury-containing protein fractions in Medaka are helpful for the understanding of the mechanisms of mercury toxicity.