酶联免疫吸附法检测鸡蛋中三聚氰胺的残留

Determination of Melamine Residues in Eggs with Enzyme Linked Immunosorbent Assay

将三聚氰胺与琥珀酸酐反应以合成半抗原,再采用混合酸酐法合成人工完全抗原,然后免疫新西兰白兔获得针对三聚氰胺的特异性抗体(IC50为42 ng/mL),以此抗体为核心建立了检测三聚氰胺的间接竞争酶联免疫吸附检测法,检测限为8.5 ng/mL。鸡蛋样品以氨化乙腈提取,正己烷去脂,微孔滤膜过滤后检测;在空白样品中添加不同浓度的三聚氰胺,回收率范围为88.9%～96.1%,变异系数小于11.6%。表明:酶联免疫吸附法,快速、简便且灵敏,可作为鸡蛋或其他动物性食品中三聚氰胺残留的常规筛选方法。

In this paper,the hapten of melamine was synthesized by coupling melamine with succinic anhydride,and the immunogen was prepared with mixed anhydride method.New Zealand rabbits were immunized with the immunogen to obtain the specific antibody toward melamine（IC50 42 ng/mL）.Then,enzyme linked immunosorbent assay based on the antibody was developed with a limit of detection at 8.5 ng/mL.Egg samples were extracted with ammoniac acetonitrile,defatted with hexane and filtered with millipore filter to be determined by the method.Recoveries from the melamine fortified blank samples were in the range of 88.9%～96.1% with coefficients of variation lower than 11.6%.The result showed that ELISA could be the routine method to determinate and screen the resdues in eggs and any other aninal food rapidly,simply and sensitivey.