Huet Anne-Catherine, Paulus Melody, Henrottin Jean, Brossard Chantal, Tranquet Olivier, Bernard Hervé, Pilolli Rosa, Nitride Chiara, Larré Colette, Adel-Patient Karine, Monaci Linda, Mills E N Clare, De Loose Marc, Gillard Nathalie, Van Poucke Christof
CER Groupe, Analytical Laboratory, Rue du Point du Jour 8, 6900, Marloie, Belgium.
INRAE, UR1268 BIA, Rue de la Géraudière, BP71627, 44316, Nantes, France.
Anal Bioanal Chem. 2022 Mar;414(8):2553-2570. doi: 10.1007/s00216-022-03912-z. Epub 2022 Feb 24.
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.
引发性测试材料的设计和生产对于食品过敏原分析方法的开发和验证至关重要。这是因为生产和加工条件以及食品基质会改变过敏原，影响其结构、可提取性和可检测性。对于旨在开发一种基于质谱的参考方法以同时准确量化两种难以分析的基质中六种致敏成分的ThRAll项目，选用了两种高度加工的基质，即巧克力棒和汤粉，在中试规模的食品生产厂中，以2、4、10和40毫克总致敏蛋白/千克食品基质的水平加入六种致敏成分（鸡蛋、牛奶、花生、大豆、榛子和杏仁）。使用食物过敏患者血清/血浆IgE验证加入成分的致敏活性，通过适当的酶联免疫吸附测定（ELISA）验证加入基质的均匀性，并评估其在4℃下至少30个月储存期的稳定性。在巧克力棒的所有添加水平下均检测到过敏原，且分布均匀，但花生和大豆仅在总致敏成分蛋白高于4毫克/千克时才可测定。汤粉的均匀性评估仅限于大豆、牛奶和花生的分析，但仍表明过敏原分布均匀。所有测试的过敏原在加入的基质中至少30个月内均保持稳定，表明它们适用于方法开发。
