超高效液相色谱串联质谱测定乳及乳制品中AFM1、AFM2

Determination of Aflatoxin M1 and M2 in Milk and Dairy Products by Ultra-high Performance Liquid Chromatography-tandem Mass Spectrometry

利用免疫亲和萃取结合超高效液相色谱-串联四极杆质谱技术(UPLC-MS/MS)建立了乳及乳制品中黄曲霉毒素M1和M2的提取、分离、确证与定量方法。乙腈和水提取样品中的黄曲霉毒素,乙腈二次去蛋白后用免疫亲和柱净化,采用Waters Acquity UPLC RHSS T3(2.1×100 mm,1.7μm)色谱柱实现分离,多反应监测(MRM)模式下以外标法定量。结果表明:黄曲霉毒素M1和M2在0.1μg/L^50μg/L范围内线性关系良好(r﹥0.998 5);检出限为0.029μg/L^0.047μg/L(S/N=3);2种黄曲霉毒素各添加水平在鲜牛奶、奶粉和奶油中的回收率为在69.56%~88.43%之间;相对标准偏差为3.69%~8.74%。

A method for the determination of 2 aflatoxins （M1,M2） in milk and dairy products by immunoaffinity extraction coupled with ultra-high performance liquid chromatography-tandem mass spectrometry （UPLC-MS/M S） was developed. Aflatoxins in samples were extracted into acetonitrile/water system and deproteinized by adding additional acetonitrile, followed by stepwise purification using an immunoaffinity column. The target compounds were then eluted with methanol. With the gradient elution using a binary mobile phase containing of 0.1%formic acid solution and methanol/acetonitrile, the two aflatoxins were separated on a Waters Acquity UPLC R HSS T3 column （2.1 ＆#215;100 mm,1.7 μm particle size）, followed by positive electrospray ionization and multi reaction monitoring provided by a triple-quadrupole tandem mass spectrometer,and quantified by external reference method. The calibration curve was linear over the concentration range of 0.1μg/L-50μg/L （r﹥0.998 5） and the detection limits of two aflatoxins were 0.029μg/L-0.047μg/L. The recoveries from milk, milk powder and cream were generally between 69.56%and 88.43%and the relative standard deviations were 3.69%-8.74%.