酶联免疫吸附分析法测定水产品及水中孔雀石绿和无色孔雀石绿

本文报道一种同时测定水产品及水样中孔雀石绿(MG)和无色孔雀石绿(LMG)的间接竞争酶联免疫吸附分析法。对无色孔雀石绿分子进行修饰,使其与载体蛋白交联,得到免疫原和包被抗原,经过多次免疫动物制得抗无色孔雀石绿的多克隆抗体。在优化的实验条件下,IC50值(标准曲线中吸光度抑制至最大吸光度值的50%时所对应的待测物浓度)为0.9~2.6μg/L,检出限为0.02~0.10μg/L,无色孔雀石绿在水样及水产品中的回收率为76.2~95.0%,与孔雀石绿的交叉反应率为95.25%。真实样品测定中,两种食用鱼养殖水样及一个鱼样中未检出孔雀石绿和无色孔雀石绿,但在观赏鱼养殖水样及另一鱼样中检出孔雀石绿和无色孔雀石,浓度分别为1.84μg/L和1.38μg/L。

Sensitive enzyme-linked immunosorbent assay for determination of malachite green and leucomalachite green in fish and fishpond water samples

An indirect competitive ELISA for the determination of malachite green and leucomalachite green in fish and fishpond water samples was developed.Leucomalachite green molecule was modified and coupled to carrier proteins to make the coating antigen and immunogen.The antibodies against LMG was obtained from rabbits.Based on the optimized conditions,IC_(50) value(concentration of analyte producing 50% of absorbance decrease in standard curve)and detection limit was found in the range of 0.9～2.6μg/L and 0.02～0.10μg/L,respectively.Recoveries of LMG in water and was found to be 95.25%.No detectable MG and LMG and was observed in one fish sample and two fish farming water samples;while the MG and LMG concentrations in one pet fishpond water sample and another fish samle were found at 1.84μg/L and 1.38μg/L ,respectively.