液相色谱串联质谱法同时测定饲料中6种代表性霉菌毒素

Simultaneous determination of 6 representative mycotoxins in feed by liquid chromatography-tandem mass spectrometry

目的建立液相色谱-串联质谱法(liquidchromatography-tandemmassspectrometry,LC-MS/MS)同时检测4种代表性饲料(猪、鸡配合饲料、玉米和麸皮)中黄曲霉毒素B1(alatoxinB1,AFB1)、呕吐毒素(deoxynivalenol, DON)、T-2毒素(T-2 toxin, T-2)、赭曲霉毒素A(ochratoxin A, OTA)、玉米赤霉烯酮(zearalenone,ZEN)、伏马毒素B1(fumonisin B1, FB1)6种霉菌毒素的方法。方法样品经乙腈-水-甲酸(80:19:1, V:V:V)震荡和超声提取,取上清液经由七水硫酸镁和C18组成的混合型吸附剂-稀释净化,采用HR-ODS色谱柱分离,以0.1%甲酸-水和5mmol/L乙酸铵-0.1%甲酸甲醇溶液作为流动相进行梯度洗脱,采用ESI正负模式转换多反应监测模式测量,基质匹配标准曲线外标法进行定量。结果在高、中、低3种添加浓度水平下, 6种霉菌毒素的平均回收率为83%~113%,相对标准偏差为3.1%~18%。结论该方法灵敏度高、精密度好、经济、环保,适用于饲料中6种代表性霉菌毒素的测定。

Objective To establish a method for the simultaneous determination of alatoxin B1(AFB1), deoxynivalenol(DON), T-2 toxin(T2), ochratoxin A(OTA), zearalenone(ZEN), fumonisin B1(FB1)in 4 representative feeds(pig, chicken compound feed, corn and bran) by liquid chromatography-tandem mass spectrometry(LC-MS/MS). Methods Samples were extracted by acetonitrile-water-formic acid(80:19:1, V:V:V) and ultrasonic extraction. The supernatant was purified by a mixture of magnesium sulfate heptahydrate and C18 adsorbent-dilution, separated by HR-ODS column using 0.1% formic acid-water and 5 mmol/L ammonium acetate-0.1% formic acid methanol solution as mobile phase for gradient elution, and ESI positive and negative mode conversion multiple reaction monitoring ion mode and matrix matching standard curve external standard method were used for quantitative analysis. Results The average recovery rates of the 6 mycotoxins were 83%–113% at the 3 levels of high, medium and low additive concentrations, and the relative standard deviations were 3.1%-18%. Conclusion This method has high sensitivity, good precision, economy and environmental protection, and is suitable for the determination of 6 representative mycotoxins in feed.