S,S-氰戊菊酯间接竞争酶联免疫吸附分析方法研究  

An Indirect Competitive Enzyme-linked Immunosorbent Assay for Esfenvalerate

作　　者：张斌[1] 王鸣华[1] 

机构地区：[1]南京农业大学植物保护学院,江苏省农药学重点实验室,农作物生物灾害综合治理教育部重点实验室,南京210095

出　　处：《分析化学》2012年第4期579-583,共5页Chinese Journal of Analytical Chemistry

基　　金：江苏高校优势学科建设工程资助项目

摘　　要：合成了S,S-氰戊菊酯的半抗原S-2-(4-氯苯基)-3-甲基丁酸-α-S-(N-丁酸基)-甲酰氨-(3-苯氧基)苄酯(Efvb)。半抗原通过混合酸酐法与卵清蛋白(OVA)偶联作为包被抗原,活泼酯法与牛血清蛋白(BSA)偶联作为免疫抗原。用免疫抗原免疫新西兰大白兔,得到抗S,S-氰戊菊酯多克隆抗体。通过异源分析及检测条件优化,确立了间接竞争酶联免疫分析方法的最佳检测条件为pH 7.4,0.4 mol/L Na+、40%甲醇-PBS溶液,建立了S,S-氰戊菊酯酶联免疫分析方法。方法的抑制中浓度(IC50)值为(3.16±0.01)mg/L;检出限(IC10)为(0.0053±0.0012)mg/L。对甲氰菊酯、溴氰菊酯、氯氰菊酯、三氟氯氰菊酯及其代谢物戊菊酸没有明显交叉反应。在自来水、河水和土壤样品中添加S,S-氰戊菊酯,其回收率分别为82.3%～108.2%,83.1%～109.2%和72.0%～91.2%。The hapten of α-S-（3-carboxy-propylcarbamoyl）-3-phenoxybenzyl-S-2-（4-Chlorophenyl）-3-methylbutyriate（Efvb） was synthesized.The haptens were conjugated with ovalbumin（OVA） as coating antigens by mixed anhydride method and with bovine serum albumin（BSA） as immunogen by active ester method.An antibody against esfenvalerate was generated by immunizing the New Zealand rabbits.After the heterologous assays and the optimization,the conditions of pH 7.4,0.4 mol/L Na＋,40% methanol were selected as the optimum assay condition for the determination of esfenvale-rate by the indirect competitive enzyme-linked immunosorbent assay（ic-ELISA）.The medium inhibitory concentration（IC50） for esfenvalerate was 3.16±0.01 mg/L,and the limit of detection（IC10） was（0.0053±0.0012） mg/L.Fenpropathrin,deltamethrin,cyhalothrin,cypermethrin and the acid of esfenvalerate metabolites have not significantly cross-reaction with antibody used in this assay.When the esfenvalerate standards were added to the tap water,river water and soil samples,the average recoveries were in the range of 82.3%-108.2%,83.1%-109.2%,72.0%-91.2%,respectively.

关 键 词：S S-氰戊菊酯 半抗原 酶联免疫吸附分析 

分 类 号：S481.8[农业科学—农药学]