A simple validated LC–MS/MS-based method was set up to detect milk contamination in bakery products,
taking the effects of food processing into account for the evaluation of allergen recovery and quantification.
Incurred cookies were prepared at eight levels of milk contamination and were cooked to expose all
milk components, including allergenic proteins, to food processing conditions. Remarkable results were
obtained in term of sufficiently low LOD and LOQ (1.3 and 4 mg/kg cookies, respectively). Precision was
calculated as intra-day repeatability (RSD in the 5–20% range) and inter-day repeatability (4 days; RSD
never exceeded 12%). The extraction recovery values ranged from 20% to 26%. Method applicability
was evaluated by analysing commercial cookies labelled either as ‘‘milk-free” or ‘‘may contain milk”.
Although the ELISA methodology is considered the gold standard for detecting allergens in foods, this
robust LC–MS/MS approach should be a useful confirmatory method for assessing and certifying
‘‘milk-free” food products.