食品科学2026，Vol.47Issue(17)：324-332,9.DOI:10.7506/spkx1002-6630-20260303-019
基于糜蛋白酶-液相色谱-串联质谱法定量烘焙食品中奶蛋过敏原
Quantification of Milk and Egg Allergens in Baked Foods Using Chymotrypsin Digestion with LC-MS/MS
摘要
Abstract
Thermal processing induces the Maillard reaction,which can specifically modify lysine and arginine residues.This modification interferes with trypsin digestion efficiency and compromises the accuracy of allergen quantification.To address this problem,we used chymotrypsin instead of trypsin for proteolysis,effectively circumventing the impact of thermal modifications.Using milk and egg as model foods,we successfully identified one highly specific and thermally stable signature peptide for each as a quantitative marker.We then established a quantitative method combining liquid chromatography-tandem mass spectrometry(LC-MS/MS)with stable isotope-labeled internal standard calibration.Method validation demonstrated high sensitivity with limits of quantification of 5 and 10 mg/kg for allergen of milk and egg in a biscuit matrix,respectively.The method exhibited excellent linearity(determination coefficient R2>0.996),reliable accuracy with spiked recoveries ranging from 73.0%to 94.7%,and good stability with precision(relative standard deviations(RSDs))below 12.6%.These results indicate that the strategy reduces the interference of thermal processing with the efficiency of enzymatic digestion and the quantitative accuracy,providing a new approach for accurate allergen quantification,risk assessment and management in complex thermally processed food matrices.关键词
食物过敏原/致敏蛋白/美拉德反应/糜蛋白酶/质谱分析Key words
food allergens/allergenic proteins/Maillard reaction/chymotrypsin/mass spectrometric analysis分类
轻工纺织引用本文复制引用
李思怡,申成英男,周美珍,赵雨萌,李熠,杨术鹏..基于糜蛋白酶-液相色谱-串联质谱法定量烘焙食品中奶蛋过敏原[J].食品科学,2026,47(17):324-332,9.基金项目
绿色食品(绿色优质农产品)过敏原识别与防控技术研究项目(GFJY20240038) （绿色优质农产品）
中国农业科学院农业科技创新工程项目(ASTIP) （ASTIP）
新疆重点研发计划项目(2022B02026-4) （2022B02026-4）
中国农业科学院基础研究中心创新计划项目(CAAS-BRC-FNH-2025-03) （CAAS-BRC-FNH-2025-03）
