苯胺嘧啶类杀菌剂嘧霉胺人工抗原的合成与鉴定

Synthesis and Identification of Pyrimethanil Artificial Antigen of Aniline Pyrimidine Fungicide

[目的]合成嘧霉胺完全抗原并加以鉴定。[方法]对嘧霉胺的结构进行设计,合成N-(4,6-二甲基嘧啶基-2-基)-1,4-苯二胺,通过与琥珀酸酐合成引入羧基得到嘧霉胺的半抗原。用混合酸酐法将半抗原与牛血清白蛋白(BSA)偶联制备免疫原,用活泼酯法将半抗原与卵清蛋白(OVA)偶联制备包被原。[结果]半抗原的结构经~1H NMR以及HPLC-MS确证。经紫外光谱分析,计算得出半抗原与牛血清白蛋白和卵清蛋白的结合比分别为14∶1和8∶1。[结论]通过嘧霉胺半抗原的合理设计,建立快速有效的嘧霉胺ELISA检测方法。

[Aims] This study aims to synthesize and identify pyrimethanil complete antigen. [Methods] N-（4,6-Dimethylpyrimidin-2-yl）benzene-1,4-diamine was synthesized, followed by introduction of the carboxyl group by reacting with succinic anhydride to obtain the hapten of the pyrimethanil. The hapten was conjugated with bovine serum albumin（BSA） by mixed acid anhydride method, and the antigen was prepared by coupling the hapten with ovalbumin（OVA） with the active ester method. [Results] The structure of the hapten was confirmed by 1H NMR and HPLC-MS. By UV spectroscopy, the ratio of hapten to bovine serum albumin and ovalbumin was calculated to be 14∶1 and 8∶1, respectively. [Conclusions] A rapid and effective method for detecting ELISA is established by the rational design of pyrimethanil hapten.