复合免疫亲和柱净化-UPLC-MS/MS测定饲料中黄曲霉毒素B_(1)、玉米赤霉烯酮和T-2毒素

Determination of aflatoxin B_(1),zearalenone and T-2 toxin in feeds by UPLC-MS/MS combined with immunoaffinity clean-up column

为建立饲料(浓缩饲料、配合饲料、植物性饲料原料)中黄曲霉毒素B_(1)、玉米赤霉烯酮和T-2毒素残留量的复合免疫亲和柱净化-UPLC-MS/MS检测方法,实验样品经86%乙腈提取,复合免疫亲和柱净化,定容后经液相快速分离,电喷雾离子化,采用MRM监测模式,正负离子扫描,外标法定量。结果表明:黄曲霉毒素B_(1)检测限为0.02μg/kg,定量限为0.05μg/kg,在0.05~2.0μg/L线性关系良好,r≥0.9990,在添加不同倍数定量限浓度水平上回收率为90.5%~97.8%,变异系数≤2.6%;玉米赤霉烯酮检测限为0.2μg/kg,定量限为0.5μg/kg,在0.5~10.0μg/L线性关系良好,r≥0.9992,在添加不同倍数定量限浓度水平上回收率为85.4%~93.7%,变异系数≤4.6%;T-2毒素检测限为0.02μg/kg,定量限为0.05μg/kg,在0.05~2.0μg/L线性关系良好,r≥0.998,在添加不同倍数定量限浓度水平上回收率为87.4%~95.2%,变异系数≤3.9%。本方法分析速度快、灵敏度高、准确可靠,可用于饲料中黄曲霉毒素B_(1)、玉米赤霉烯酮和T-2毒素的检测分析。

A new method for determination of aflatoxin B_(1),zearalenone and T-2 in feeds by ultra performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS)was developed.The samples were extracted using the solution of 86%acetonitrile,and purified through immunoaffinity clean-up column,rapid separation by liquid phase after constant volume.The identification and quantification of the target compounds were carried out by positive and negative electrospray ionization(ESI±)in the multiple reaction monitoring(MRM)mode using external standard method.The results showed that the limits of detection(LOD)and the limits of quantification(LOQ)for aflatoxin B_(1) were 0.02μg/kg and 0.05μg/kg.The calibration curves showed good linearity in the range of 0.05~2.0μg/L(r≥0.9990).The recovery rates were90.5%~97.8%at different concentration levels,with the relative standard deviations(RSD)less than or equal to 2.6%.The limits of detection(LOD)and the limits of quantification(LOQ)for zearalenone were 0.2μg/kg and 0.5μg/kg.The calibration curves showed good linearity in the range of 0.5~10.0μg/L(r≥0.9992).The recovery rates were 85.4%~93.7%at different concentration levels,with the relative standard deviations(RSD)less than or equal to 4.6%.The limits of detection(LOD)and the limits of quantification(LOQ)for T-2 toxin were 0.02μg/kg and 0.05μg/kg.The calibration curves showed good linearity in the range of 0.05~2.0μg/L(r≥0.998).The recovery rates were 87.4%~95.2%at different concentration levels,with the relative standard deviations(RSD)less than or equal to 3.9%.The method was simple,rapid,sensitive,accurate,reliable for the determination of aflatoxin B_(1),zearalenone and T-2 toxin in feeds.