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目的建立免疫亲和柱净化-HPLC光化学柱后衍生荧光检测法,用于测定植物油中黄曲霉毒素B_1、B_2、G_1、G_2。方法样品用乙腈-水(20+80,体积比)混合液提取,提取液经免疫亲和柱净化,净化液经C18色谱柱分离,用光化学柱后衍生器衍生、荧光检测器进行检测,采用外标法进行定量。结果在优化的条件下,4种黄曲霉毒素能达到很好的分离效果,黄曲霉毒素B_1、G_1在0.2~20.0 ng/ml范围内线性关系良好,检出限为0.04μg/kg;黄曲霉毒素B_2、G_2在0.1~10.0 ng/ml范围内线性关系良好,检出限为0.02μg/kg,4种黄曲霉毒素的回收率为82.5%~95.3%,RSD为2.4%~5.9%。结论该方法快速、简单、准确、灵敏、重现性好,是测定植物油中的黄曲霉毒素B_1、B_2、G_1、G_2的可靠方法。
OBJECTIVE To establish an immunoaffinity column purification-HPLC fluorescence post-column derivatization fluorescence detection method for the determination of aflatoxins B_1, B_2, G_1, G_2 in vegetable oils. Methods The sample was extracted with a mixture of acetonitrile and water (20 + 80, volume ratio), and the extract was purified by immunoaffinity column. The purified liquid was separated on a C18 column and derivatized with a photochemical post-column derivatior. The fluorescence detector was used for detection. External standard method for quantitative. Results Under the optimized conditions, the four aflatoxins could achieve good separation effect. The aflatoxins B_1 and G_1 had a good linearity in the range of 0.2-20.0 ng / ml with a detection limit of 0.04 μg / kg. The linear relationship between toxins B_2 and G_2 in the range of 0.1 ~ 10.0 ng / ml was good with a detection limit of 0.02 μg / kg. The recoveries of four aflatoxins were 82.5% -95.3% with RSDs of 2.4% -5.9%. Conclusion The method is rapid, simple, accurate, sensitive and reproducible. It is a reliable method for the determination of aflatoxins B_1, B_2, G_1, G_2 in vegetable oils.