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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.
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| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
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