基于高分辨质谱技术的婴幼儿食品中过敏原蛋白质的高灵敏检测

摘要

基于高效液相色谱-串联质谱(LC-MS/MS)技术,选择稳定性好、灵敏度高的特征肽段,利用平行反应监测(PRM)技术,实现了多类过敏原蛋白质的高灵敏度同时检测,并成功应用于婴幼儿食品中过敏原成分的分析.对于婴幼儿食品中蛋白质的提取,与传统的丙酮沉淀法比,采用膜上原位样品预处理方法(i-FASP)可实现更高的蛋白质提取效率和抗干扰能力.所检测的过敏原蛋白质的定量限(LOQ)最小可达到0.028 mg/L,其线性范围最宽可跨越4个数量级,且线性关系良好(相关系数R2≥0.99).该方法为食品中过敏原蛋白质组学快速分析提供了一种可靠的分析方法.%A novel method of the simultaneous detection of multiple kinds of allergenic proteins in infant food with parallel reaction monitoring (PRM)mode using liquid chromatography-tan-dem mass spectrometry (LC-MS/MS)was established. In this method,unique peptides with good stability and high sensibility were used to quantify the corresponding allergenic proteins. Furthermore,multiple kinds of allergenic proteins are inspected simultaneously with high sensi-tivity. In addition,such method was successfully used for the detection of multiple allergenic proteins in infant food. As for the sample preparation for infant food,compared with the tradi-tional acetone precipitation strategy,the protein extraction efficiency and capacity of resisting disturbance are both higher with in-situ filter-aided sample pretreatment (i-FASP)method. All allergenic proteins gave a good linear response with the correlation coefficients (R2 )≥0. 99, and the largest concentration range of the allergenic proteins could be four orders of magni-tude,and the lowest detection limit was 0. 028 mg/L,which was better than that reported in references. Finally, the method was conveniently used to detect the allergens from four imported infant food real samples. All the results demonstrate that this novel strategy is of great significance for providing a rapid and reliable analytical technique for allergen proteomics.