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Development of incurred chocolate bars and broth powder with six fully characterised food allergens as test materials for food allergen analysis

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Abstract

The design and production of incurred test materials are critical for the development and validation of methods for food allergen analysis. This is because production and processing conditions, together with the food matrix, can modify allergens affecting their structure, extractability and detectability. For the ThRAll project, which aims to develop a mass spectrometry–based reference method for the simultaneous accurate quantification of six allergenic ingredients in two hard to analyse matrices. Two highly processed matrices, chocolate bars and broth powder, were selected to incur with six allergenic ingredients (egg, milk, peanut, soy, hazelnut and almond) at 2, 4, 10 and 40 mg total allergenic protein/kg food matrix using a pilot-scale food manufacturing plant. The allergenic activity of the ingredients incurred was verified using food-allergic patient serum/plasma IgE, the homogeneity of the incurred matrices verified and their stability at 4 °C assessed over at least 30-month storage using appropriate enzyme-linked immunosorbent assays (ELISA). Allergens were found at all levels from the chocolate bar and were homogenously distributed, apart from peanut and soy which could only be determined above 4 mg total allergenic ingredient protein/kg. The homogeneity assessment was restricted to analysis of soy, milk and peanut for the broth powder but nevertheless demonstrated that the allergens were homogeneously distributed. All the allergens tested were found to be stable in the incurred matrices for at least 30 months demonstrating they are suitable for method development.

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Acknowledgements

The present article is the sole responsibility of the authors. The positions and opinions presented in this article are those of the authors alone and do not necessarily represent the views of any official position or scientific works of EFSA. EFSA guidance documents and other scientific outputs of EFSA can be found in the EFSA website, http://www.efsa.europa.eu. Bert Verlinden, Jari Van De Vijver, Hans Steurbaut, Jurgen Baert, Stijn Degroote and Estelle De Paepe are acknowledged for their practical assistance in the production of the test materials. Emilie Perrin is acknowledged for her technical assistance in IgE binding experiments, Amandine Lamote and Paolo Paque for the analyses of stability by ELISA, Martin Mailleux for the analyses by mass spectrometry. We gratefully thank Gianfranco Mamone and the Luigia di Stasio Institute of Food Sciences – National Research Council, Avellino, Italy – for providing us with the almond and hazelnut reference materials.

Funding

This work has received financial support from the European Food Safety Authority (EFSA) for the ThRAII project GP/EFSA/AFSCO/2017/03. It was also co-funded by the UK Food Standards Agency (FS101209), the Grant FS101206 (UK FSA) for the development of quality control materials for food allergen analysis and the Belgian Federal Agency for the Safety of the Food Chain (FASFC).

Sera or plasma from patients allergic to peanuts, milk, egg, almond, hazelnut or soy was obtained from the Biological Resource Center (BB-0033–00038) of Clinical Immunology and Allergy Service of Angers University Hospital (France) with the informed consent of the patients.

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Authors and Affiliations

  1. CER Groupe, Analytical Laboratory, Rue du Point du Jour 8, 6900, Marloie, Belgium

    Anne-Catherine Huet, Melody Paulus, Jean Henrottin & Nathalie Gillard

  2. INRAE, UR1268 BIA, Rue de la Géraudière, BP71627, 44316, Nantes, France

    Chantal Brossard, Olivier Tranquet & Colette Larré

  3. INRAE, Aix-Marseille University, Biodiversité Et Biotechnologie Fongiques (BBF), UMR1163, 13009, Marseille, France

    Olivier Tranquet

  4. INRAE-CEA, Service de Pharmacologie Et d’Immunoanalyse, Laboratoire d’Immuno-Allergie Alimentaire, Bât 136-CEA Saclay, 91191, Gif-sur-Yvette, France

    Hervé Bernard & Karine Adel-Patient

  5. Institute of Sciences of Food Production, CNR-ISPA, Bari, Italy

    Rosa Pilolli & Linda Monaci

  6. School of Biological Sciences, Division of Infection, Immunity and Respiratory Medicine, Manchester Academic Health Science Centre, Manchester Institute of Biotechnology, The University of Manchester, Manchester, UK

    Chiara Nitride & E. N. Clare Mills

  7. Flanders Research Institute for Agriculture, Fisheries and Food (ILVO), Burgemeester Van Gansberghelaan 115, 9820, Merelbeke, Belgium

    Marc De Loose

  8. Flanders Research Institute for Agriculture, Fisheries and Food (ILVO), Brusselsesteenweg 370, 9090, Melle, Belgium

    Christof Van Poucke

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  1. Anne-Catherine Huet
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Correspondence to Anne-Catherine Huet.

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Huet, AC., Paulus, M., Henrottin, J. et al. Development of incurred chocolate bars and broth powder with six fully characterised food allergens as test materials for food allergen analysis. Anal Bioanal Chem 414, 2553–2570 (2022). https://doi.org/10.1007/s00216-022-03912-z

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  • DOI: https://doi.org/10.1007/s00216-022-03912-z

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