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本研究以邻苯二甲酸二甲酯(DMP)为研究目标,以4-氨基邻苯二甲酸二甲酯为半抗原,通过重氮化法偶联载体蛋白并免疫动物,制备针对DMP的特异性兔多克隆抗体。通过棋盘滴定法和单因素实验确定最佳的实验参数,即包被原浓度为50μg/L,一抗浓度为92.5μg/L,二抗浓度为1μg/m L,药物稀释液为p H 6.0的纯水,竞争反应时间为40 min,基于此建立了间接竞争化学发光酶联免疫分析法检测DMP。本方法对DMP的检出限为0.29μg/L,线性检测范围为0.74~30.32μg/L,与13种结构和功能类似物的交叉反应率均<5%,通过倍比稀释降低白酒、酱油样品基质干扰,对样品的平均回收率在80.2%~116.0%之间,平均相对标准偏差<3.6%,结果与GC-MS法的测定结果相符。本方法适用于食品样品中DMP的快速检测。
In this study, dimethyl phthalate (DMP) as the research goal, with 4-amino-dimethyl phthalate as a hapten, diazotization method by coupling the carrier protein and immunized animals, prepared for DMP-specific Rabbit polyclonal antibody. The optimal experimental parameters were determined by chessboard titration and single factor experiments. The original concentration of coating was 50μg / L, the concentration of primary antibody was 92.5μg / L, the concentration of secondary antibody was 1μg / ml, the dilution of drug was p H 6.0 Of pure water. The competitive reaction time was 40 min. Indirect competition chemiluminescence enzyme-linked immunosorbent assay was used to detect DMP. The detection limit of this method for DMP was 0.29μg / L, the linear range was 0.74 ~ 30.32μg / L, the cross-reaction rate with all 13 structures and functional analogues was less than 5% The sample matrix interference, the average recovery of the sample was between 80.2% and 116.0%, the average relative standard deviation was less than 3.6%. The results were in good agreement with the results of GC-MS. The method is suitable for the rapid detection of DMP in food samples.