液相色谱-串联质谱法同时测定面粉中5种镰刀菌毒素方法的建立

Simultaneous determination of 5 fusarium toxins in wheat flour by ultra performance liquid chromatography-tandem mass spectrometry

目的建立同时检测面粉中DON、15A—DON、3A—DON、ZEN、NIV5种镰刀菌毒素的超高效液相色谱-串联质谱（UPLC—MS／MS）方法。方法用乙腈～水（84：16）溶液提取样品，经净化柱净化后，氮气吹干，20％甲醇液定容，用UPLC—MS／MS进行测定。UPLC—MS／MS检测镰刀菌毒素的条件为梯度洗脱，流动相为0．3％氨水和乙腈；质谱采集模式为电喷雾正离子多反应监测模式。结果5种镰刀菌毒素在5～1 000μg／L线性范围内呈线性关系良好（相关系数r值均〉O．99）；检出限：DON、3A—DON、15A—DON均为0．4 μg／kg，ZEN为0．3／μg／kg，NIV为0．2／μg／kg。不同加标水平的5种镰刀菌毒素回收率为70％～105％，相对标准偏差（RSD）为5．2％～8．9％。结论UPLC—MS／MS法具有操作简单、灵敏度高、重现性好等特点，可同时检测面粉中5种镰刀菌毒素。

Objectives To develop a simutaneous determination of 5 fusarium toxins （DON, 15A-DON, 3-DON , ZEN,NIV5） in wheat flour by ultra performance liquid chromatography-tandem mass spectrometry（UPLC-MS/MS）. Methods Thesamples were extracted by 84% （v/v） acetonitrile-water mixure, followed by purification by purification column and dried bynitrogen. Using 20% （v/v） methanol-water mixture to dissolve to samples to a certain volume ,which was analyzed by UPLC-MS/MS with gradient elution. Acetonitrile and 0. 3% （v/v）ammonia-water were used as the mobile phases. The mass spectro-metric acquisitions were carried out by multiple reaction monitoring（MRM） in electro spray positive ionization mode. ResultsThe 5 fusarium toxins showed good linear relationships in concentration ranges of 5-1 000/μg/L with the correlation coefficientsmore than 0.99. The limits of detection（LOD） for DON,3A-DON,15A-DON were all 0.4 μg/kg, LOD of ZEN and NIV were0.3 /μg/kg and 0.2 μg/kg, respectively. The recovery rates of 5 fusarium toxins at the different spiking levels ranged from 70%-105% ,with relative standard deviations （RSD） of 5.2%-8.9% Conclusion The established UPLC-MS/MS method is easyto operate,with high sensitivity and reproducibility. It satisfies requirement for simultaneous determination of 5 fusarium toxinsin wheat flour.