直接稀释-超高效液相色谱-串联质谱法快速测定谷物及其制品中16种真菌毒素

A dilute-and shoot approach using ultra high-performance liquid chromatograph-mass/mass spectrometry for 16 mycotoxins analysis in cereals and products

目的建立谷物及其制品中黄曲霉毒素、赭曲霉毒素、单端孢霉烯族类毒素、伏马菌素等16种真菌毒素的超高效液相色谱-串联质谱(UPLC-MS/MS)检测方法。方法 5 g样品加入20 ml乙腈-水-甲酸(79∶20∶1,V/V)溶液提取,10 000 r/min离心5 min后稀释,Cortecs C_(18)色谱柱(100 mm×3.0 mm,1.6μm)分离,以乙腈-0.1%甲酸水作为流动相梯度洗脱,采用四极杆串联质谱仪电喷雾模式检测,同位素稀释内标法定量。结果 16种真菌毒素在线性范围内(0.05~200 ng/ml)具有良好的线性关系,相关系数(r^2)>0.99,方法定量限为0.1~50μg/kg,加标回收率为72.67%~126.92%之间,相对标准偏差(RSD)为0.15%~16.83%。应用本方法分析英国弗帕斯检测技术研究所(FAPAS)质控样品,测定结果均在标示范围内,方法准确度良好。结论本方法具有良好的灵敏度、回收率和重复性,前处理方法简单,适用于常规实验室对谷物及其制品中真菌毒素的检测,满足日常真菌毒素监测工作的需要。

Objective To develop and validate the ultra high-performance liquid chromatograph coupled with mass spectrometry（ UPLC-MS/MS） method for simultaneous determination of 16 mycotoxins,including aflatoxins,ochratoxins,trichothecenes,fumonisins and so on. Methods Five grams of the sample was extracted with acetonitrile/water/formic acid（ 79 ∶ 20 ∶ 1,V/V）. After centrifuged and diluted,the extraction was separated by Cortecs C_（18）（ 100 mm × 3. 0 mm,1. 6 μm） analytical column under acetonitrile-0. 1 % formic acid gradient eluting,then was determined by UPLC-MS/MS.Dilution isotope internal calibration was used for qualification. Results The result showed good linearities（ 0. 05-200 ng/ml） for 16 mycotoxins in the certain correlation ranges with the coefficients all above 0. 99. The limits of quantification ranged from 0. 1 to 50 μg/kg. The average recoveries of 16 mycotoxins at three spiked levels ranged in72. 67 %-126. 92 % with relative standard deviations of 0. 15 %-16. 83 %. The accuracies were acceptable by detecting natural matrix quality control samples. Conclusion The method is rapid,simple,sensitive,reproducible and accurate for simultaneous determination of multiple mycotoxins in cereals and products in routine laboratories. It meets the the requirement of mycotoxins monitoring in relative food.