摘要
为检测环境和农产品中乙草胺的含量,建立间接竞争酶联免疫检测法（ELISA）。以乙草胺和3-巯基丙酸为原料合成半抗原,与牛血清蛋白偶联制备免疫原,免疫新西兰大白兔获得了多克隆抗体;以间接竞争性ELISA法（ic-ELISA）考察了多克隆抗体的特异性并优化了影响该方法的3种因素（甲醇浓度、离子强度和pH值）;在此基础上测定了乙草胺在多种基质中的添加回收率。结果显示：该方法具有很高的灵敏度和很强的特异性,最低检测限为0.024 4 mg/L,与结构相似的农药无明显交叉反应;在自来水、田水、土壤、玉米和大豆样品中的添加浓度为0.5~10.0 mg/L或0.1~1.0 mg/kg,回收率为73.2%~117.6%,变异系数为1.7%~9.2%,符合农药残留分析要求。优化的ELISA检测方法适合环境和农产品中乙草胺的检测。
An indirect competitive enzyme-linked immunsorbent assay（ic-ELISA） was developed to detect acetochlor in environmental samples and agro-product.Acetochlor was used to react with 3-mercaptopropionic acid to synthesize the hapten,acetochlor-3-mercaptopropionic acid（AMPA）,which was covalently conjugated with the carrier proteins bovine serum albumin（BSA） as immunogen.Polyclonal antibodies against acetochlor were raised by immunizing New Zealand White rabbits with hatpen-BSA.The specificity of polyclonal antibodies was tested by ic-ELISA and the influence of several factors including the concentration of methanol,ionic strength and pH value were also evaluated to improve the sensitivity of the assay.The optimized ELISA was shown to have a high sensitivity and specificity for acetochlor,with a limit of detection（IC10） of 0.024 4 mg/L and low cross-reactivities towards structurally rlated compoinds（pretilachlor,butachlor,metazachlor,metalaxyl）.The recoveries of acetochlor added in tap water,soil water,soil,maize,and soybean in three replicates for all measurements were in the range of 73.2%-117.6% at additive concentraions between 0.5 mg/L and 10.0 mg/L（or 0.1-1.0 mg/kg）,with the relative standard deviatons（RSD） being 1.7%-9.2%.The accuracy and precision well matched requirements of residue analysis.The optimized ic-ELISA provides an alternative analytical method for the determinzation of acetochlor in environmental and food samples.
出处
《江苏农业学报》
CSCD
北大核心
2011年第2期420-425,共6页
Jiangsu Journal of Agricultural Sciences
基金
国家“863”项目(2006AA108447)
