超高效液相色谱-电喷雾质谱法检测花生过敏原Ara h 2

Identification and quantification of peanut allergen Ara h 2 using ultra-performance liquid chromatography-tandem mass spectrometry

目的建立烘培食品中花生过敏原Ara h 2的液相质谱联用定量检测方法。方法选择花生蛋白中的致敏蛋白Ara h 2作为目标蛋白,筛选出该致敏蛋白的特异肽,人工合成特异肽标准品和特异肽内标,从而建立直接检测花生致敏蛋白Ara h 2的准确定量方法。同时还对全国不同地区的20种花生中致敏蛋白Ara h 2的含量进行检测分析,初步统计得出致敏蛋白Ara h 2和花生蛋白的换算系数,并以Ara h 2作为生物标记物检测10种烘培食品中花生蛋白的残留量。结果花生样品中致敏蛋白Ara h 2的定量限为4.45μg/g,回收率在106.0%~107.8%之间。烘培食品中,定量限可达到6.23μg/g,回收率在107.0%~113.2%之间。结论本方法特异性强、灵敏度高、定量准确,具有良好的应用前景。

Objective To establish a reliable ultra-high-performance liquid chromatography-mass spectrometry method to determine the peanut allergen Ara h 2 in roasted food.Methods Peanut allergen Ara h 2 was quantified by filtrating their signature peptides,the synthetic specific peptides standard and its internal standard.Current validated method was successfully applied to the determination of the peanut allergen Ara h 2 content in 20 kinds of peanut in different regions.And the results showed the conversion coefficient of peanut allergen Ara h 2 and peanut protein.Thus,Ara h 2 was chosen as biomarker to detect peanut protein in 10 kinds of roasted food.Result The limit of quantitation with peanut allergen Ara h 2 in peanut samples was 4.45μg/g and the recoveries were 106.0%~107.8%.In roasted food,the limit of quantitation was 6.23 μg/g and recoveries were in the range of 107.0%~113.2%.Conclusion Compared to the previous methods,the developed approach is a highly specific,sensitive and accurate method,which has a good application prospect.