免疫亲和柱净化-大体积流通池高效液相色谱法同时检测食品中6种黄曲霉毒素

Simultaneous Determination of Six Aflatoxins in Food by Immunoaffinity Purification-high Performance Liquid Chromatography Combined with a Large Volume Flow Cell

建立了免疫亲和柱净化-大体积流通池无需衍生同时测定食品中6种黄曲霉毒素(Aflatoxins)的检测方法。样品采用乙腈-水(84:16,V/V)超声辅助提取,用免疫亲和柱净化,经过XBridge TM C18柱(150 mm×4.6 mm,5μm)分离。采用乙腈-甲醇-水(15:25:60,V/V)为流动相,大体积流通池荧光检测器检测,无需衍生,外标法定量。6种黄曲霉毒素在9.5 min内有效分离,从样品前处理到结果分析整个过程小于50 min。黄曲霉毒素B1、B2、G1、G2、M1、M2的检出限(LOD,S/N=3)分别为0.05μg/kg、0.02μg/kg、0.05μg/kg、0.02μg/kg、0.04μg/kg、0.03μg/kg,能够满足我国对食品中黄曲霉毒素限量的要求。6种黄曲霉毒素标准曲线线性良好,线性相关系数r2大于0.999;样品加标回收率为77.6%~90.5%,精密度RSD为2.42%~6.08%。本方法简便、准确、灵敏度高,无需衍生即可同时检测食品中6种黄曲霉毒素,适用于食品中6种黄曲霉毒素的快速定量测定。

A method without derivative based on immunoaffinity purification and fluorimetric detection combined with large volume flow cell was developed for simultaneous determination of six kinds of aflatoxins in food. The samples were ultrasonic-extracted with acetonitrile-aqueous solution(84:16, V/V), cleaned up by immunoaffinity column, and then were separated by XBridge TM C18 column(150 mm×4.6 mm, 5μm). The analytes were detected by large volume flow cell fluorescence detection in a mobile phase of acetonitrile-aqueous solution(84:16, V/V) without derivatization. External standard method was used for quantitative. All analytes were successfully separated within 6 min, and one sample determination from pretreatment to result analysis was finished within 50 min. The detection limit(LOD, S/N=3) of aflatoxin B1, B2, G1, G2, M1, M2 respectively were 0.05 μg/kg, 0.02 μg/kg, 0.05 μg/kg, 0.02 μg/kg, 0.04 μg/kg, 0.03 μg/kg, meeting the national limits of aflatoxins in food. The correlation coefficient of six kinds of aflatoxins, r2,were more than 0.999, and the spiked recoveries were in the ranges of 77.6%~90.5%, and relative standard deviation(RSD, n=3) were 2.42%~6.08%. The method was simple, effective and sensitive, detecting the aflatoxins without derivatization and suitable for the simultaneous determination of six kinds of aflatoxins in food.