To protect the allergic consumer, analytical methods need to be capable of detecting allergens in fnished products that typically contain multiple allergens. An LC/MS/MS method for simultaneous detection of seven allergens was developed and compared with commercially available ELISA kits. The detection capabilities of this novel method were demonstrated by analyzing incurred material containing milk, egg, soy, peanut, hazelnut, walnut, and almond. Bread was chosen as a model matrix. To assess the influence of baking on the method’s performance, analysis was done before and after baking. The same samples were analyzed with ELISA test kits from ELISA Systems, Morinaga, Neogen, and r-Biopharm. Peanut, hazelnut, walnut, and almond could be detected with both ELISA and LC/MS/MS regardless of whether the product was baked or not. LC/MS/MS clearly showed superior detection of milk in processed matrixes compared to ELISA, which exhibited signifcantly lower sensitivities when analyzing the baked products. Similar results were obtained when analyzing egg; however, one kit was capable of detecting egg in the processed samples as well.

Background Effective allergenic risk assessment and management are important to limit the use of precautionary statements such as ‘may contain’ and to be able to protect allergic consumers. However, such approaches require reliable analytical tools for the detection of allergens in food. Very few validation data are available for the comparison of results obtained with different allergen detection methods. This is certainly due to the lack of harmonized validation protocols and of recognized reference materials. Aims The Monitoring and Quality Assurance Working Group on Food Allergens will provide incurred reference materials with egg and milk proteins at various concentrations. Materials and Methods The development of an incurred reference material for the analysis of milk and egg allergens in a baked cookie food matrix is described. Results and Discussion We present the results of the development of the incurred reference material and a pre‐ring trial with two incurred reference materials for milk detection methods: cookies and soy‐based infant formula. Conclusions The material produced seems to be suitable as reference material as well as for testing the performance of test kits. The forthcoming validation study according to the harmonized validation protocol will significantly and positively impact on future validation procedures. Dumont V, Kerbach S, Poms R, Johnson P, Mills C, Popping B, Tömösközi S & Delahaut P (2010). Development of milk and egg incurred reference materials for the validation of food allergen detection methods. Quality Assurance and Safety of Crops & Foods, 2, 208–215

Celiac disease and wheat allergy are the most common adverse reactions triggered by cereal proteins, mainly gluten, which is one of the 14 allergenic food ingredients that must be labeled on food products in the European Union (EU). To meet the requirements of this regulation, reliable analytical methodology for proper quantification of gluten is necessary. However, validation of presently used methods (ELISA and lateral flow device) is limited partly due to the lack of reference methods and incurred reference materials. To solve this problem, the goal of our work was to develop an incurred reference material for the quantification of gluten under the auspices of EU-FP6 funded Network of Excellence MoniQA. During this work, we produced a processed model product (cookie) containing gliadin (major allergenic fraction of gluten) in a defined amount. This paper addresses the development process of this material together with the associated problems (insufficient homogeneity and low recovery) and their solutions. As a result, an incurred food matrix was produced on a laboratory-scale with a potential use as a reference material. The model product was tested by an ELISA method followed by a comparative study of commercially available ELISA kits to investigate the applicability of the product. Preliminary results of this study are also presented.

The management of food allergens involves several stakeholders including food manufacturers, consumers, enforcement authorities and analytical laboratories. The MoniQA Working Group ''Food Allergens'' and representatives of these stakeholder groups develop a synthesis of their needs and requirements, identify gaps and suggest ways forward to address these gaps. Analysis for food allergens is an essential adjunct to hazard and risk management procedures within the food industry. These analyses are important for enforcement authorities who require reliable methods in support of both the specific food labelling legislation and the more general food safety legislation. Furthermore, analytical methods are essential in the validation/verification of cleaning protocols, which help to reduce levels of potentially contaminating allergen to one that is no longer harmful for the majority of the allergic consumers. Nevertheless, there is a general lack of validated and robust analytical methodology for analysis of most food allergens. On the one hand, universally recognized reference materials are missing, on the other hand there is an urgent need to harmonise validation protocols at an international level. Currently, members of the MoniQA WG ''Food Allergens'' together with the AOAC Allergen Community are developing a validation protocol for allergen ELISAbased test kits helping to provide standardised methods with known acceptance criteria for the users as well as for the method developers.The Working Group will seek to develop the initiative required to address these issues, in collaboration with other organisations e.g. the EuroPrevall project, FARRP, AOAC and the CEN WG12 Food Allergens over the coming years. 50

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