UPLC-MS/MS法同时检测食品中3种主要牛奶过敏原

Simultaneous Detection of Three Major Milk Allergens in Food by UPLC-MS/MS

为准确定量食品中的牛奶过敏原,采用超高效液相色谱-串联质谱法同时检测3种主要牛奶过敏原β-乳球蛋白、αs1-酪蛋白和αs2-酪蛋白,避免单一致敏蛋白在加工中的降解和灵敏度不高导致的假阴性。标准蛋白酶解液经纳升液相色谱-串联轨道阱高分辨质谱分析后,筛选得到8条特征肽段。利用三重四极杆质谱多反应监测模式,选择面粉作为空白基质进行方法学验证。结果表明,该方法在1.6~30000 ng/mL范围内线性良好,R2>0.999;定量限分别为β-乳球蛋白50μg/g、αs1-酪蛋白3.2μg/g、αs2-酪蛋白40μg/g;平均回收率为86.41%~98.60%,相对标准偏差不大于8.95%;基质效应在86.04%~97.48%之间。此方法可应用于含牛奶和不含牛奶的实际商品,旨在鉴别食品中过敏原标签与产品实际过敏原间的差异,降低牛奶过敏消费者的健康隐患。

An ultra-high performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS)method was developed to simultaneously detect the three major milk allergensβ-lactoglobulin,αs1-casein andαs2-casein in food,which could avoid false negatives caused by factors such as the degradation of single allergens during processing and low sensitivity.The standard protein hydrolysates were analyzed by nanoflow liquid chromatography-tandem orbitrap high resolution mass spectrometry,obtaining eight signature peptides.Wheat flour was selected as the blank matrix for methodological validation in the multi-reaction monitoring(MRM)mode by triple quadrupole mass spectrometry.This method showed a good linear relationship in the concentration range of 1.6–30000 ng/mL with correlation coefficient(R2)>0.999.The limits of quantitation(LODs)ofβ-lactoglobulin,αs1-casein,andαs2-casein were 50,3.2,and 40μg/g,respectively.The average recoveries were between 86.41%and 98.60%with relative standard deviations(RSDs)≤8.95%,and the matrix effects were 86.04%–97.48%.This method could be applied to actual products with and without milk in order to identify whether there is any inconsistence between the allergen label and the actual allergens in food,and more broadly to reduce the potential health risks for milk-sensitive consumers.