酶联免疫法检测混合植物油中黄曲霉毒素B1的含量

Determination of aflatoxin B1 in mixed vegetable oil by enzyme immunosorbentassay

中文关键词:  黄曲霉毒素B1  直接竞争酶联免疫吸附法  混合植物油

英文关键词:Aflatoxin B1  direct competitive enzyme-linked immunosorbent assay  mixed vegetable oil

中文摘要:

      摘 要: 目的 建立一种测定混合植物油中黄曲霉毒素B1含量方法。方法 采用直接竞争酶联免疫吸附法。样品经甲醇溶液提取后, 提取的抗原与黄曲霉毒素B1标记物与微孔板上的黄曲霉毒素B1特异性单克隆抗体竞争结合, 洗涤除去未结合杂质。将底物加入孵育, 产生蓝色产物, 加入终止液终止反应蓝色产物变成黄色产物, 450nm处测量吸光度, 与样品中含量成反比。结果 本方法在2h内完成混合植物油中黄曲霉毒素B1含量的测定。黄曲霉毒素B1的加标浓度在1.0ug/kg/mL-20.0ug/kg之间时的回收率为76.5%-120.2%, 方法定量限为1.00 μg/kg。结论 该方法快速、准确、灵敏, 适合测定混合植物油中黄曲霉毒素B1。

英文摘要:

      ABSTRACT: Objective To establish a method for determination of aflatoxin B1 in mixed vegetable oil. Methods The direct competitive enzyme-linked immunosorbent assay (ELISA) was used. After the sample was extracted by methanol, the extracted antigen was combined with aflatoxin B1 labeled with aflatoxin B1 specific monoclonal antibody on the microplate, and then washed to remove the unbound impurity. When the substrate was added to incubation, the blue product was produced, and the termination reaction was stopped. The blue product became yellow product, and the absorbance at 450nm was inversely proportional to the content in the sample. Results The method was applied to determine aflatoxin B1 content in mixed vegetable oil within 2h. When the spiked concentration of aflatoxin B1 was between 1.0ug/kg/mL-20.0ug/kg, the recovery was 76.5%-120.2%, and the limit of quantification was 1 g/kg. Conclusion The method is rapid, accurate and sensitive. It is suitable for the determination of aflatoxin B1 in mixed vegetable oil.