精恶唑禾草灵酶联免疫吸附分析方法研究  

An Enzyme-linked Immunosorbent Assay for Detection of Fenoxaprop-p-ethyl

作　　者：王俊东[1] 施海燕[1] 叶永浩[1] 王鸣华[1] 

机构地区：[1]南京农业大学植物保护学院,南京210095

出　　处：《分析化学》2009年第1期82-86,共5页Chinese Journal of Analytical Chemistry

基　　金：国家863项目(No.2006AA108447);贵州大学绿色农药与农业生物工程教育部重点实验室开放基金(No.2008GDGP0102)项目

摘　　要：建立了测定精恶唑禾草灵的间接竞争酶联免疫吸附分析方法(ic-ELISA),合成了精恶唑禾草灵的半抗原精恶唑禾草酸(hapten)。通过碳二亚胺法将精恶唑禾草酸交联于牛血清蛋白(BSA)作为免疫抗原,通过活泼酯法将精恶唑禾草酸交联于卵清蛋白(OVA)作为包被抗原,Hapten-BSA为免疫原制备了精恶唑禾草灵的兔抗血清,间接非竞争酶联免疫吸附分析方法测得其效价达1.024×105。确定了0.3mol/LNa+强度的磷酸盐缓冲液(pH7.5)和10%的甲醇为精恶唑禾草灵间接竞争酶联免疫吸附分析方法的最佳工作条件。本方法的IC50为(0.084±0.012)mg/L,检出限(IC10)为(0.0064±0.002)mg/L。对大部分芳氧苯氧基丙酸酯类除草剂,如精喹禾灵、氰氟草酯、高效吡氟甲禾灵没有明显的交叉反应,对于恶唑酰草胺有一定的交叉反应,这与两者的结构有关。To develop a competitive indirect enzyme-linked immunosorbent assay （ci-ELISA） for fenoxaprop- p-ethyl, a hapten （fenoxaprop acid） was synthesized. Hapten conjugated to bovine serum albumin （BSA） with the carbodiimide method was used as immunogens and ovalbumin （OVA） with mixed anhydride method was used as the coating antigens. The New Zealand rabbits were immunized by conjugate of Hapten-BSA and titres of anti-fenoxaprop-p-ethyl serum （1.024 × 10^5） were determined by non-competitive indirect enzyme-linked assay procedure. After optimization of the ic-ELISA conditions, such as pH values, inonic strengths, concentrations of methanol, pH 7.5, PBS of 0.3 mol/L Na^＋ , and 10% methanol were determined as optimum assay conditions. The ICs0 for fenoxaprop-p-ethyl was （0. 084 ＋ 0. 012 ） mg/L, and lower detection limit （LDL） was （0.0064 ＋0.002） mg/L. The cross-reactivities of other aryloxyphenoxypropionic acid herbicides to the antibodies were less than 0.1% in the assays, except metamifop because of its structures.

关 键 词：精恶唑禾草灵 多克隆抗体 酶联免疫吸附分析 

分 类 号：TQ450.263[化学工程—农药化工]