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

中文关键词:  花生过敏原  液相色谱-串联质谱法  Ara h 2  特异肽

英文关键词:peanut allergens  liquid chromatography-mass spectrometry  Ara h 2  specific peptide

中文摘要:

      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4.45 μg/g, 回收率在106.0%~107.8%之间。烘培食品中, 定量限可达到6.23 μg/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。

英文摘要:

      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.