Lee J K, Ahn K C, Park O S, Kang S Y, Hammock B D
Department of Agricultural Chemistry, Chungbuk National University, Cheongju 361-763, Korea.
J Agric Food Chem. 2001 May;49(5):2159-67. doi: 10.1021/jf001140v.
A competitive enzyme-linked immunosorbent assay (ELISA) for the chloronicotinyl insecticide imidacloprid was developed using a polyclonal antibody produced against a hapten conjugated through the imidazolidine to keyhole limpet hemocyanin. In the standard curve of imidacloprid, an IC(50) of 17.3 ng/mL was obtained using a competitive heterologous system at pH 10. Very low cross-reactivity was found for some structurally related compounds including the insecticide thiacloprid. The high cross-reactivity with a metabolite containing the carbonyl group in the imidazolidine moiety suggests the involvement of its polarity and stereochemical fitness in forming the antibody--antigen complex. The effects of various assay conditions, including organic solvents, detergent content, salt concentration, and pH on the sensitivity were evaluated. High-performance liquid chromatography was run for comparison to validate the ELISA with fortified water samples, the correlation being 0.997-0.998 (n = 15) with a slope of 1.10--1.38. The ELISA turned out to be a convenient tool for monitoring imidacloprid residues in agricultural and environmental samples.
开发了一种用于氯代烟碱类杀虫剂吡虫啉的竞争性酶联免疫吸附测定（ELISA）方法，该方法使用针对通过咪唑烷与钥孔血蓝蛋白偶联的半抗原产生的多克隆抗体。在吡虫啉的标准曲线中，使用pH 10的竞争性异源系统获得的IC（50）为17.3 ng/mL。对于一些结构相关的化合物，包括杀虫剂噻虫啉，发现其交叉反应性非常低。与咪唑烷部分含有羰基的代谢物具有高交叉反应性，这表明其极性和立体化学适应性在形成抗体-抗原复合物中起作用。评估了各种测定条件，包括有机溶剂、洗涤剂含量、盐浓度和pH对灵敏度的影响。运行高效液相色谱进行比较，以验证加标水样的ELISA，相关性为0.997-0.998（n = 15），斜率为1.10-1.38。ELISA结果证明是监测农业和环境样品中吡虫啉残留的便捷工具。
