全自动免疫亲和柱净化-液相色谱串联质谱法同时测定饼粕类饲料原料中黄曲霉毒素

Simultaneous Determination of Aflatoxins in Oil Cake Feedstuff by Fully Automated Immunoaffinity Column Purification Combined with LC-MS/MS

研究旨在建立全自动免疫亲和柱净化-液相色谱串联质谱(LC-MS/MS)同时测定饼粕类饲料原料中黄曲霉毒素B1、B2、G1、G2(AFB1、AFB2、AFG1、AFG2)含量的分析方法。通过比较不同提取条件和净化方式对黄曲霉毒素(AFs)检测结果的影响,确定较佳前处理方法。样品经40%乙腈提取、磷酸盐缓冲液稀释,采用自动免疫亲和柱净化,得到的样品溶液使用LC-MS/MS进行定量分析。基于上述方法对花生粕、豆粕、棉籽粕等饲料原料样品进行分析测定,结果表明:AFs在0.02~10.00 ng/mL范围内线性关系良好(R2≥0.9950),方法定量限为0.1μg/kg,平均回收率为82.2%~109.8%,RSD≤8.9%。该方法可批量自动化处理样品,提高工作效率,避免因人员操作导致的结果偏差,适合于饲料原料中多种AFs的精准测定。

An analytical method for simultaneous determination of Aflatoxin B1,B2,G1,G2(AFB1,AFB2,AFG1,AFG2)in oil cake feedstuff by fully automated immune affinity purification combined with LC-MS/MS was established in this experi⁃ment.The optimization conditions of sampling,concentration and purification for sample pretreat⁃ment were studied,respectively.The samples were extracted with 40%acetonitrile,diluted with phosphate buffer,and then purified by automated immunoaf⁃finity column system.The obtained solutions were determined by LC-MS/MS finally.Based on these methods,different oil cake feedstuff samples were tested.The results showed that good linearity was ob⁃tained in the ranges of 0.02 ng/mL to 10.00 ng/mL,with linear coefficients(R2)higher than 0.9950.The limits of quantification(LOQ)were 0.1μg/kg,Average recoveries from the fortification samples ranged be⁃tween 82.2%and 109.8%,with relative standard deviations(RSD)≤8.9%.The new method is suitable for batch automatic sample processing,improving efficiency,avoiding deviations caused by personnel op⁃eration,and is appropriate for accurate determination of aflatoxins in oil cake feedstuff.