Milk allergy is the most common food allergy presented in early childhood which in some cases may persist into adulthood as well. Proteins belonging to casein and whey fractions of milk trigger an allergic response in susceptible individuals. Milk is present as an ingredient in many foods and it can be also present as casein or whey enriched milk-derived ingredients. As whey proteins are more susceptible to thermal processing than caseins, conventional methods often posed a challenge in accurate detection of whey allergens, particularly from a processed complex food matrix. In this study we have developed a targeted mass spectrometry method to detect the presence of both casein and whey allergens from thermally processed foods. A pool of 19 candidate peptides representing four casein proteins and two whey proteins was identified using a discovery-driven target selection approach from various milk-derived ingredients. These target peptides were evaluated by parallel reaction monitoring of baked cookie samples containing known amounts of nonfat dry milk. The presence of milk could be detected from baked cookies incurred with NFDM at levels as low as 1 ppm using seven peptides representing α, β, and κ casein proteins and three peptides representing whey protein, β-lactoglobulin by this consensus PRM method.

Data links to manuscript figures: Figure 1, generated from data in skyline file "Milk Ingredient_Discovery Analysis". Figure 2, 3, & S4, generated from data in skyline file "PRM of candidate targets in milk-derived ingredients". Figure 4 & S5, generated from data in skyline files "Method sensitivity assay_in presence of gelatin" and "Method sensitivity assay_in absence of gelatin". Figure 5 & S6, generated from data in skyline file "Milk detection from NFDM incurred cookie-dough samples".