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目的建立普洱茶中黄曲霉毒素B1(AFB1)的免疫亲和柱净化-酶联免疫吸附检测方法。方法粉碎后的普洱茶样品经三氯甲烷提取,水浴挥干后用甲醇-PBS溶解,样品液通过黄曲霉毒素B1免疫亲和柱净化洗脱,在450nm波长使用酶联免疫试剂盒(ELISA)定量检测黄曲霉毒素B1含量。结果该方法在2.0~100.0μg/kg浓度范围内百分吸光率和浓度对数呈良好的线性关系,相关系数达0.999 8,样品最低检出限为1.0μg/kg,重复测定相对标准偏差为4.72%,3个水平(1.0、10.0、50.0μg/kg)的样品加标回收率为83.00%~104.90%。结论本法具有简单、快速、高效等优点,适用于普洱茶复杂样品的AFB1定量检测。
Objective To establish a method for the determination of aflatoxin B1 (AFB1) in Pu-erh tea by immunoaffinity column-enzyme-linked immunosorbent assay. Methods The pulverized Pu’er tea samples were extracted with chloroform and evaporated in a water bath and then dissolved in methanol-PBS. The sample solution was eluted by aflatoxin B1 immunoaffinity column and eluted at 450 nm with enzyme-linked immunosorbent assay (ELISA) Quantitative detection aflatoxin B1 content. Results The method showed a good linear relationship between the percentage absorbance and the logarithm of concentration in the concentration range of 2.0-100.0μg / kg, the correlation coefficient was 0.999 8, the minimum detectable limit of the sample was 1.0μg / kg, the relative standard deviation was 4.72%. The recoveries of three spiked samples (1.0, 10.0, 50.0 μg / kg) were 83.00% -104.90%. Conclusion This method is simple, rapid and efficient. It is suitable for the quantitative determination of AFB1 in complex samples of Pu’er tea.