液相色谱-串联质谱法测定虾饲料中7种真菌毒素

建立了虾饲料中黄曲霉毒素B_1(AFB_1)、黄曲霉毒素M_1(AFM_1)、T-2毒素(T-2)、HT-2毒素(HT-2)、脱氧雪腐镰刀菌烯醇(DON)、赭曲霉毒素A(OTA)和玉米赤霉烯酮(ZEN)的液相色谱-串联质谱(LC-MS/MS)同时检测方法。样品中加入15 mL乙腈-水(体积比4∶1)和10 mL乙腈饱和正己烷,涡旋混匀,超声提取,MycoSpin^TM400多毒素净化柱净化后上机测定。采用Hypersil Gold (150 mm×2.1 mm,5μm)色谱柱进行分离,以甲醇-水(含0.03%氨水)为流动相梯度洗脱,在电喷雾电离模式下正、负离子同时扫描检测,基质匹配外标法定量。在优化条件下,7种真菌毒素的线性相关系数(r^2)均大于0.991,检出限为1.83～12.63μg/kg。在高、中、低3个加标水平下,各目标毒素的回收率为87.5%～116%,相对标准偏差为2.4%～18%。该法可为水产饲料中多种真菌毒素的同时检测提供参考。

Determination of 7 Mycotoxins in Shrimp Feed by Liquid Chromatography-Tandem Mass Spectrometry

A liquid chromatography-tandem mass spectrometric(LC-MS/MS) method was developed for the quantitative determination of aflatoxin B1(AFB1),aflatoxin M1(AFM1),T-2 toxin(T-2),HT-2 toxin(HT-2),deoxynivalenol(DON),ochratoxin A(OTA) and zearalenone(ZEN) in shrimp feed.Samples were extracted with 15 mL acetonitrile-water(4 ∶1,by volume),and degreased with 10 mL acetonitrile-saturated hexane.The extract was cleaned up with a MycoSpinTM400 multitoxin LC-MS/MS clean-up column.The target compounds were separated on an Hypersil Gold(150 mm×2.1 mm,5 μm)chromatographic column by gradient elution with methanol-water containing 0.03%ammonia as mobile phase,and analyzed simultaneously by electron spray ionization mass spectrometry in positive or negative ion mode,and quantified by the matrix-matched external standard method.Under the optimized conditions,there were good linearities for the analytes in corresponding concentration range with their correlation coefficients(r2) more than 0.991.The limits of detection(LOD)ranged from 1.83 μg/kg to 12.63 μg/kg.Average recoveries for the target mycotoxins at high,medium and low spiked levels ranged from 87.5%to 116%with relative standard deviations of 2.4%-18%.The developed method could provide a reference for the simultaneous determination of mycotoxins in aquatic feeds.