液相色谱-三重串联四极杆质谱测定粮油中的黄曲霉毒素

Determination of aflatoxins in cereals and oils by liquid chromatography-triple quadrupole tandem mass spectrometry

建立了超声提取-液相色谱-电喷雾三重串联四极杆质谱测定玉米、大米、大豆等粮油固体样品中黄曲霉毒素B1、B2、G1和G2(AFB1、AFB2、AFG1和AFG2)的方法。分析前对样品进行超声提取,优化得到最佳超声提取条件:溶剂为甲醇-水(含40 g/L NaCl)(80∶20,v/v)溶液、料液比为1∶3(g∶mL)、温度为50℃、时间为3 min。然后对提取的样品进行免疫亲和特异性净化。最后与液相色谱-电喷雾三重串联四极杆质谱联用,使用C18反相色谱柱,流动相为甲醇-10 mmol/L乙酸铵水溶液梯度洗脱,以黄曲霉毒素M1(AFM1)作为内标进行定量测定。结果表明,AFB1、AFB2、AFG1和AFG2的检出限分别为0.002、0.004、0.004和0.012μg/kg。方法的加标回收率为87%～111%,日内相对标准偏差(RSD)和日间RSD分别不大于6.7%和5.6%。实验结果表明该方法可以有效地降低基质效应的影响,相比于外标法能极大地提高方法的准确度。

The high performance liquid chromatography（HPLC）-triple quadrupole tandem mass spectrometry was applied for the determination of the aflatoxins： B1（AFB1）,B2（AFB2）,G1（AFG1） and G2（AFG2）,in cereals and oils.The samples were first extracted by ultrasonic method.The optimized conditions of ultrasonic extraction were as follows： temperature of 50 ℃,extraction time of 3 min, methanol-water（containing 40 g/L NaCl）（80∶20,v/v） solution as the medium and a ratio of sample to solvent of 1∶3（g∶mL）.The extracts were then purified using an immunoaffinity column.The separation was performed on a C18 column with mobile phases of 10 mmol/L ammonium acetate and methanol in gradient elution.The sensitive detection of the four AFT compounds by electrospray ionization mass spectrometry（ESI-MS） was carried out in selected reaction monitoring（SRM） mode with aflatoxin M1（AFM1） as the internal standard.Under the optimized conditions,the limits of detection of AFB1,AFB2,AFG1 and AFG2 were 0.002,0.004,0.004 and 0.012 μg/kg,respectively.The recoveries of aflatoxins in different spiked cereals and oils were in the range from 87% to 111%.The intraday and inter-day precisions were not more than 6.7% and 5.6%,respectively.In comparison with the external standard method,this method can effectively inhibit the matrix effects,and greatly improve the accuracy.