El-Gendy Kawther, Mosallam Eman, Ahmed Nabila, Aly Nagat
Department of Pesticide Chemistry, Faculty of Agriculture, Alexandria University, Alexandria, Egypt.
Department of Mammalian Toxicology, Central Agricultural Pesticide Lab, Agricultural Research Center, Alexandria, Egypt.
Anal Biochem. 2018 Sep 15;557:1-6. doi: 10.1016/j.ab.2018.07.004. Epub 2018 Jul 10.
A sensitive linker-assisted enzyme-linked immunosorbent assay (L'ELISA) was developed for the analysis of glyphosate in Egyptian soil samples. Polyclonal glyphosate antibodies were produced from rabbits immunized with glyphosate protein conjugate. The conjugate was prepared by activating the carboxylic groups of proteins; thyroglobulin or bovine serum albumin with 1-ethyl-3- (3-diaminopropyl) carbodiimide hydrochloride and N-hydroxysulfosuccinimide followed by directly coupled to the amino group of glyphosate. The L'ELISA used succinic anhydride to derivatize glyphosate, which mimics the epitopic attachment of glyphosate to thyroglobulin. L'ELISA recognized the derivatized glyphosate with a limit of detection (LOD) of 0.8 ng g and sensitivity (IC value) of 0.018 μg g. The recovery values of the spiked soil samples with different concentrations of glyphosate were in the range of 87.4-97.2%. Good correlation was achieved between L'ELISA and conventional high-pressure liquid chromatography (HPLC) with fluorescence detection. This study demonstrated the utility and convenience of the sensitive, simple, practical and cost-effective L'ELISA method for glyphosate analysis in soil samples. Also, it is ideal for rapid screening of a large number of environmental samples.
开发了一种灵敏的连接物辅助酶联免疫吸附测定法（L'ELISA）用于分析埃及土壤样品中的草甘膦。用草甘膦蛋白偶联物免疫兔子制备多克隆草甘膦抗体。通过用1-乙基-3-（3-二氨基丙基）碳二亚胺盐酸盐和N-羟基磺基琥珀酰亚胺活化蛋白质（甲状腺球蛋白或牛血清白蛋白）的羧基，然后直接与草甘膦的氨基偶联来制备偶联物。L'ELISA使用琥珀酸酐衍生化草甘膦，其模拟草甘膦与甲状腺球蛋白的表位附着。L'ELISA识别衍生化的草甘膦，检测限（LOD）为0.8 ng/g，灵敏度（IC值）为0.018 μg/g。不同浓度草甘膦加标土壤样品的回收率在87.4-97.2%范围内。L'ELISA与常规高压液相色谱（HPLC）荧光检测之间具有良好的相关性。本研究证明了灵敏、简单、实用且经济高效的L'ELISA方法在土壤样品草甘膦分析中的实用性和便利性。此外，它非常适合大量环境样品的快速筛查。
