Objective To establish a detection method for soybean allergens in plant-based protein products based on ultra performance liquid chromatography-tandem mass spectrometry, and conduct research on the quantification and authenticity determination of allergy protein in plant-based protein products. Methods After the soybean protein isolate was hydrolyzed by trypsin, the data were collected by high resolution mass spectrometry and matched with the Uniprot database to screen out the soybean specific allergen peptides. The response surface design was adopted to optimize the extraction conditions of the allergen protein, thereby constructing the quantitative method of allergens. Actual sample tests were conducted on commercially available plant-based protein products to preliminarily obtain the content of soybean allergy proteins in different plant-based protein products. Results The 6 kinds of soybean quantitative peptides with good linearity and correlation coefficients above 0.995 were selected. The limits of detection and the limits of quantification of the method were 0.182–2.08 μg/g and 0.607–6.920 μg/g. The spiked recoveries were 97.1%–126.6% and the intra-day and inter-day precisions were 5.17%–10.20% and 5.02%–11.70%. Conclusion This method has strong specificity and high sensitivity, and can well detect plant-based protein products, achieving accurate quantification of soybean allergens without isotope-labeled peptide segments. It has a good application prospect in the detection of allergen in plant-based protein products. Meanwhile, it can achieve the identification of adulteration in plant-based protein products and the discrimination of allergic information on food labels.
关键词
植物替代蛋白产品、大豆过敏蛋白、超高效液相色谱-串联质谱法、特征多肽、响应面优化
KeyWord
plant-based proteins; soybean allergens proteins; ultra performance liquid chromatography-tandem mass spectrometry; specific peptide; response surface optimization
该研究建立了基于超高效液相色谱-串联质谱法用于植物替代蛋白产品中大豆过敏蛋白的检测,在植物替代蛋白产品的过敏蛋白检测方面具有较好的应用潜力, 也为植物替代蛋白产品的掺假鉴别及食品标签的过敏信息判别提供了技术支撑。
该研究针对植物替代蛋白产品大豆过敏蛋白检测与真实性判定的行业痛点,建立超高效液相色谱 - 串联质谱检测方法,选题贴合食品安全监管与过敏人群健康保障需求,应用价值明确。实验设计严谨,通过高分辨质谱结合数据库筛选出 6 种高特异性大豆定量多肽,经响应面优化提取条件,方法学验证显示相关系数>0.995,检出限、回收..