Institute of Sciences of Food Production, CNR-ISPA, Bari, Italy.
Flanders Research Institute for Agriculture, Fisheries and Food, Melle, Belgium.
Food Chem. 2021 May 1;343:128533. doi: 10.1016/j.foodchem.2020.128533. Epub 2020 Nov 4.
Peptide marker identification is an important step in development of a mass spectrometry method for multiple allergen detection, since specificity, robustness and sensitivity of the overall analytical method will depend on the reliability of the proteotypic peptides. As part of the development of a multi-analyte reference method, discovery analysis of two incurred food matrices has been undertaken to select the most reliable peptide markers. Six allergenic ingredients (milk, egg, peanut, soybean, hazelnut, and almond) were incurred into either chocolate or broth powder matrix. Different conditions of protein extraction and purification were tested and the tryptic peptide pools were analysed by untargeted high resolution tandem mass spectrometry and the resulting fragmentation spectra were processed via a commercial software for sequence identification. The analysis performed on incurred foods provides both a prototype effective and straightforward sample preparation protocol and delivers reliable peptides to be included in a standardized selected reaction monitoring method.
肽标记物的鉴定是开发用于多种过敏原检测的质谱方法的重要步骤，因为整体分析方法的特异性、稳健性和灵敏度将取决于蛋白特征肽的可靠性。作为多分析物参考方法开发的一部分，对两种受污染的食品基质进行了发现分析，以选择最可靠的肽标记物。将六种过敏原成分（牛奶、鸡蛋、花生、大豆、榛子和杏仁）掺入巧克力或肉汤粉基质中。测试了不同的蛋白质提取和纯化条件，并通过靶向高分辨率串联质谱分析胰蛋白酶肽池，然后通过商业软件对所得的片段图谱进行序列鉴定。对受污染食品进行的分析提供了一种原型有效且直接的样品制备方案，并提供了可靠的肽，以便包含在标准化选择反应监测方法中。
