单克隆抗体间接竞争ELISA测定水体中氟虫腈

Determination of fipronil in water by ELISA based on monoclonal antibody

制备了氟虫腈单克隆抗体 ,在此基础上建立了水体中氟虫腈测定的间接竞争性酶联免疫吸附测定 (ELISA)法 .在优化条件下 ,氟虫腈测定的线性浓度范围为 1 0 -1 μg·L-1 ～ 1 0 3μg·L-1 ,最低检出浓度 (I1 0 )为 0 0 8μg·L-1 ,检测灵敏度 (I50 )为6 6 8μg·L-1 ;常见的几种与氟虫腈结构类似的农药的交叉反应率均低于 1 % ,不干扰氟虫腈的检测 ;2 0 %丙酮、5 %甲醇、2 %二甲基甲酰胺、2 %乙腈和 2 %乙酸乙酯基本不影响免疫测定 .利用建立的间接竞争性ELISA方法检测不同水样中氟虫腈 ,结果表明不同水样均可直接进行测定 。

An indirect competitive enzyme|linked immunosorbent assay (ELISA) was developed for the determination of fipronil in water using a high|affinity mono c lonal antibody. With the sera of highest specificity, the antigen|coated indire c t ELISA showed an I_(50) of 6.68?μg·L^(-1) with a detection limit of 0 .08?μg·L^(-1).The antibody showed negligible cross-reactivity with fipronil-related compounds, i nc luding chlorfenapyr, flufenoxuron, chlorfluazuron, cyhalothrin, trifluralin, whi ch makes the assay suitable for the selective detection of fipronil. The assay w as unaffected by several organic solvents when under lower concentration, i.e. , acetone <20%, methanol <5%, and DMF, ethanol, and ethyl acetate <2%, respecti vely. Fipronil was spiked at 10, 1 and 0.1?μg·L^(-1) to water samples to evaluate the reliability of the immunoassay. The recovery of fipronil for tap, mineral, purified, pool, river, sea, and paddy field water, as well as groundwater were 9 4.7%～110%, 83.0%～103.6%, 86.0%～98.6%, 94.0%～100.2%, 105.6%～110.0%, 82.0%～116.0%, 87. 0%～107.2%, and 92.0%～106.0%, respectively. The assay was initially applied to the analysis of fipronil in environmental waters.