FOOD CHEMICAL CONTAMINANTS Selecting Processing Robust Markers Using High-Resolution Mass Spectrometry for the Detection of Milk in Food Products Kaatje Van Vlierberghe , 1 Maxime Gavage , 2,3 Marc Dieu , 3 Patsy Renard , 3 Thierry Arnould , 3 Nathalie Gillard , 2 Katleen Coudijzer , 1 Marc de Loose , 1 Kris Gevaert , 4,5 and Christof Van Poucke 1, * 1 ILVO Flanders Research Institute for Agriculture, Fisheries, and Food, Technology and Food Science Unit, Brusselsesteenweg 370, BE-9090 Melle, Belgium, 2 CER Groupe, Rue du Point du Jour 8, 6900 Marloie, Belgium, 3 University of Namur, Laboratory of Biochemistry and Cell Biology (URBC)-Namur Research Institute for Life Sciences (NARILIS), 61, rue de Bruxelles, 5000 Namur, Belgium, 4 VIB-UGent Center for Medical Biotechnology, Technologiepark-Zwijnaarde 75, BE-9052 Ghent, Belgium, 5 Ghent University, Department of Biomolecular Medicine, Technologiepark-Zwijnaarde 75, BE-9052 Ghent, Belgium *Corresponding author’s e-mail: christof.vanpoucke@ilvo.vlaanderen.be Abstract Background: Cow’s milk allergy is one of the most reported food allergies in Europe. To help patients suffering from food allergies it is important to be able to detect milk in different foods. An analytical method that is gaining interest in the field of allergen detection is ultrahigh performance liquid chromatography-tandem mass spectrometry, where the analyte is a target peptide. When these peptide biomarkers are selected, the effect of food processing should be taken into account to allow a robust detection method. Objective: This work aims at identifying such processing stable peptide markers for milk for the ultrahigh performance liquid chromatography-tandem mass spectrometry based detection of food allergens in different food products. Method: Milk-incurred food materials that underwent several processing techniques were produced. This was followed by establishing tryptic peptide profiles from each matrix using ultrahigh performance liquid chromatography-high resolution mass spectrometry. Results: A careful comparison of peptide profiles/intensities and the use of specific exclusion criteria resulted in the selection of eight peptide biomarkers suitable for application in ultrahigh performance liquid chromatography-tandem mass spectrometry based milk detection methods. One of these markers is an a-lactalbumin specific peptide, which has been determined to be stable in different incurred materials for the first time. Conclusions: To our knowledge, this is the first systematic and experimentally based approach for the selection of suitable milk peptide biomarkers robust toward multiple, often applied food processing techniques for milk. Ensuring the exact knowledge of the food processing circumstances by starting from well-defined raw material and using fully controlled Received: 10 August 2021; Revised: 14 October 2021; Accepted: 6 November 2021 VC The Author(s) 2021. Published by Oxford University Press on behalf of AOAC INTERNATIONAL. All rights reserved. For permissions, please email: journals.permissions@oup.com 463 Journal of AOAC INTERNATIONAL, 105(2), 2022, 463–475 https://doi.org/10.1093/jaoacint/qsab147 Advance Access Publication Date: 15 November 2021 Research Article Downloaded from https://academic.oup.com/jaoac/article/105/2/463/6428662 by guest on 19 July 2023