基于大豆过敏蛋白的植物替代蛋白产品真实性研究

Study on the authenticity determination of plant-based proteins based on soybean allergens

中文关键词:  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

英文关键词:plant-based proteins  soybean allergens proteins  ultra performance liquid chromatography-tandem mass spectrometry  specific peptide  response surface optimization

中文摘要:

      目的 基于超高效液相色谱-串联质谱法建立植物替代蛋白产品中大豆过敏蛋白的检测方法, 开展植物替代蛋白产品中过敏蛋白的定量和真实性判定研究。方法 大豆分离蛋白经胰蛋白酶水解后, 通过高分辨质谱采集数据并与Uniprot数据库匹配, 筛选出大豆特异性过敏蛋白多肽, 并采用响应面设计优化过敏蛋白提取条件, 从而构建过敏原定量方法。对市售植物替代蛋白产品进行实际样品检测, 初步得到不同植物替代蛋白产品中大豆过敏蛋白含量。结果 筛选出线性较好的6种大豆定量多肽、相关系数均大于0.995, 方法检出限为0.182~2.080 μg/g、方法定量限为0.607~6.920 μg/g, 回收率为97.1%~126.6%, 日内精密度为5.17%~10.20%, 日间精密度为5.02%~11.70%。结论 本方法特异性强、灵敏度高, 可以较好地针对植物替代蛋白产品进行检测, 实现了在无同位素标记肽段的情况下对大豆过敏蛋白的准确定量。在植物替代蛋白产品的过敏蛋白检测上具有较好的应用前景, 同时可实现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。

英文摘要:

      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.