Abstract: An ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method was established for the detection of pine nut allergen Pin k 2 in food matrices. Pine nuts were ground, degreased, and enzymatically extracted and the hydrolysate was separated and analyzed by using an Easy-nLC 1000-QExecutive high-resolution mass spectrometer, and the mass spectral data obtained were processed using Protein Pilot TM software and the Uniprot protein database. The specificity was verified by Basic Local Alignment Search Tool (BLAST), and three pine nut-specific peptides were selected finally. The developed method exhibited a good linear relationship in the concentration range of 0.001–50 mg/mL, and the limit of quantification was 1 mg/kg. The average recoveries for blank biscuit, chocolate and beverage were 88.50%–107.57%, with a relative standard deviation (RSD) not exceeding 6.08%, and the matrix effect was 89.77%–96.13%. This method has the advantages of high sensitivity and good specificity, and can be applied to the detection of pine nut allergens in food samples such as biscuits, chocolate, and beverages, which provides technical support for the authentication of food labels and the detection of latent allergens in foods.

Key words: pine nut; allergen; ultra-high performance liquid chromatography-tandem mass spectrometry method; detection