研究论文
余向阳,颜春荣,刘贤进.单克隆抗体间接竞争ELISA测定水体中氟虫腈[J].环境科学学报,2004,24(5):910-914
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
- 基金项目:水利部“948”(975157)项目;国家“863”(2001AA246031)项目
- 余向阳
- 江苏省农业科学院食品质量安全与检测研究中心, 南京210014
- 颜春荣
- 江苏省农业科学院食品质量安全与检测研究中心, 南京210014
- 刘贤进
- 江苏省农业科学院食品质量安全与检测研究中心, 南京210014
- 摘要: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10-1μg·L-1~103μg·L-1, 最低检出浓度(I10)为0.08μg·L-1, 检测灵敏度(I50)为6.68μg·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;20%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, 方法的精密度和准确度均符合残留测定的要求.
- Abstract: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 I50 of 6.68μμg·L-1 with a detection limit of 0.08μμg·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 0.1μμg·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 94.7%~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.
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