Food allergy is an increasing health problem due to its growing prevalence worldwide. European Commission Regulation No.1169/2011 established the list of 14 allergenic ingredients (and by-product) whose presence must be indicated in the food labels. Current legislation does not address the accidental contamination of allergens due to cross-contamination occurring along the entire food chain therefore, the risk of allergen contamination is likely to exist. Targeted LC-MS/MS-based methods demonstrated relevant advantages for allergen analysis. Linda Monaci (CNR-ISPA, Bari, Italy) and Rosa Pilolli (CNR-ISPA, Bari, Italy) are part of the ThRAll project funded by EFSA for the development of a reference MS-based method for the multiple allergen detection in food matrices. In this webinar, they show methods with optimized sample preparation and liquid chromatography separation coupled with laminar flow tandem mass spectrometry (MS) for the detection and quantitation of milk and egg allergens in in processed food products.