Development and validation of a matrix independent calibration strategy for targeted mass spectrometry-based quantification of milk protein allergens from complex food matrices

ABSTRACT: Quantitative detection of food allergens is important in assessing the risk of eliciting an allergic response in susceptible individuals in the event of an accidental exposure and to ensure compliance with allergen management and labeling regulations. Targeted mass spectrometry-based quantitative methods provide an opportunity to analyze allergens from complex food matrices which have undergone food processing. However, probable interferences of diverse food matrices mandates preparation of matrix-matched calibration curves, challenging method adaptation to multiple matrices. In this study, we have demonstrated that a novel matrix-independent calibration strategy employing an exogenous carrier protein as background in food matrices and calibrants, enables quantitative detection of amounts of milk allergen at concentrations relevant for risk assessment applications. Matrix-independent calibration curves prepared with egg white powder in background yielded comparable quantitative values with a matrix-matched calibration curve. Performance of matrix-independent calibration strategy with egg white powder was evaluated for nonfat dry milk (NFDM) incurred baked cookie, beverages containing pasteurized milk, and casein-incurred chocolate. Peptides representing both casein and whey fractions of milk were detectable from concentrations as low as 1 ppm NFDM in cookie matrix (46-99% recovery across the peptides), 0.5 ppm total milk protein (TMP) in pasteurized beverages (94-138% recovery), or 1 ppm casein in chocolate (35-51% recovery). Although the quantitative accuracy was low from traditionally difficult chocolate and baked cookie matrices, variance in quantification was within 30% across the inter-day replicates indicating method robustness for high throughput analysis of samples required in testing labs.

ORGANISM(S): Bos Taurus

SUBMITTER: Melanie Downs  

PROVIDER: PXD040564 | panorama | Mon Apr 08 00:00:00 BST 2024

REPOSITORIES: PanoramaPublic