Quantifying allergenic proteins using antibody‐based methods or liquid chromatography–mass spectrometry/mass spectrometry: A review about the influence of food matrix, extraction, and sample preparation

Abstract Accurate quantification of allergens in food is crucial for ensuring consumer safety. Pretreatment steps directly affect accuracy and efficiency of allergen quantification. We systematically reviewed the latest advances in pretreatment steps for antibody‐based methods and liquid chromatography–mass spectrometry/mass spectrometry (LC–MS/MS) protein quantification methods in food. For antibody‐based methods, the effects induced by food matrix like decreased allergen solubility, epitope masking, and nonspecific binding are of the upmost importance. To mitigate interference from the matrix, effective and proper extraction can be used to obtain the target allergens with a high protein concentration and necessary epitope exposure. Removal of interfering substances, extraction systems (buffers and additives), assistive technologies, and commercial kits were discussed. About LC–MS/MS quantification, the preparation of the target peptides is the crucial step that significantly affects the efficiency and results obtained from the MS detector. The advantages and limitations of each method for pre‐purification, enzymatic digestion, and peptide desalting were compared. Additionally, the application characteristics of microfluidic‐based pretreatment devices were illustrated to improve the convenience and efficiency of quantification. A promising research direction is the targeted development of pretreatment methods for complex food matrices, such as lipid‐based and carbohydrate‐based matrices.