复合免疫亲和柱净化-液相色谱-串联质谱法测定动物源食品中6种黄曲霉毒素和6种玉米赤霉醇类真菌毒素残留量

Determination of Aflatoxins and Zeranols in Animal-Originated Foods by High Performance Liquid Chromatography-Tandem Mass Spectrometry Combined With Immunoaffinity Clean-up Column

建立了动物源食品（猪肉、鱼肉、猪肝）中6种黄曲霉毒素（AFB1、AFB2、AFG1、AFG2、AFM1和AFM2）和6种玉米赤霉醇类真菌毒素（α-玉米赤霉醇、β-玉米赤霉醇、α-玉米赤霉烯醇、β-玉米赤霉烯醇、玉米赤霉酮和玉米赤霉烯酮）残留量的复合免疫亲和柱净化-高效液相色谱-串联质谱（HPLC-MS/MS）检测方法。样品经β-葡萄糖苷酸/硫酸酯复合酶酶解后,用甲醇-乙腈（20∶80,V/V）提取,提取液经玻璃纤维滤纸过滤,滤液用PBS溶液稀释,复合免疫亲和柱富集和净化后,采用HPLC-MS/MS法分析。12种目标分析物中AFB2和AFG2的线性范围为0.03~6.0μg/L,其余目标分析物的线性范围为0.05~20μg/L,线性相关系数均大于0.999,检出限在0.01~0.03μg/kg范围内,定量限在0.04~0.09μg/kg范围内。分别以0.5,1.0和5.0μg/kg添加浓度水平进行方法学验证,平均回收率为73.6%~98.4%,相对标准偏差（RSD）为1.9%~11.2%。本方法简便、灵敏,能够满足动物源食品中痕量黄曲霉毒素和玉米赤霉醇类真菌毒素残留的测定要求。

A high performance liquid chromatography-tandem mass spectrometric （ HPLC-MS / MS） method coupled with an immunoaffinity clean-up column was successfully developed for determination of aflatoxins （AFB1 , AFB2 , AFG1 , AFG2 , AFM1 and AFM2 ） and zeranols （ α-zeranol, β-zeranol, α-zearalenol,β-zearalenol, zearalanone and zearalenone ）. The sample was extracted with methanol-acetonitrile （20∶ 80, V/ V） after enzymatic digestion by β-glucuronidase / sulfatase, and the extraction solution was passed through＆amp;nbsp;glassy fiber filter paper and then diluted with phosphate buffer solution （PBS）. The reconstituted solution was cleaned up with IAC-AZ immunoaffinity column, and then analyzed by HPLC-MS / MS in multiple reaction monitoring （MRM） mode. The results indicated that the linear detection range was 0. 03-6. 0 μg / L for AFB2 and AFG2 , and 0. 05-20 μg / L for the rest compounds. The correlation coefficients were above 0. 999. The limits of detection （LOD） and limits of quantitation （LOQ） were 0. 01-0. 03 μg / kg and 0. 04-0. 09 μg / kg, respectively. The recoveries of the aflatoxins and zeranols were in the range of 73. 6% -98. 4% at the spiked levels of 0. 5, 1 and 5 μg / kg, and the relative standard deviations （RSDs） were in the range of 1. 9% -11. 2% . The method was proved to be simple and accurate, and suitable for the rapid determination of aflatoxins and zeranols in animal-originated foods.