超高效液相色谱-电喷雾质谱法检测花生 过敏原Ara h 2

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。

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

Peanut allergen Ara h 2 was quantified by employing the synthetic internal standard based on the molarequivalent relationship among the internal standard, Ara h 2 and their signature peptides.A reliable ultra-high-performance liquid chromatography-mass spectrometry method for determination of peanut allergen Ara h 2 was developed. Compared to the previous methods, the developed approach with mass spectrometry operated in multiple reaction monitoring mode offered a highly specific, sensitive and accurate method. The established method was extensively validated by determining sensitivity (limit of quantitation 4.45 ug/g) and recovery (106.0%~107.8%). Current validated method was successfully applied to the peanut allergen Ara h 2 content in 20 kinds of peanut from Chinese 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 biomarkers to detect peanut protein in 10 kinds of roasted food.And the established method was extensively validated by determining sensitivity (limit of quantitation 6.23 ug/g) and recovery (107.0%~113.2%).