A Novel Approach for the Detection of Potentially Hazardous Pepsin Stable Hazelnut Proteins as Contaminants in Chocolate-Based Food

• Published in: Journal of agricultural and food chemistry Vol. 52; no. 25; pp. 7726 - 7731
• Main Authors: Akkerdaas, Jaap H, Wensing, Marjolein, Knulst, André C, Stephan, Oliver, Hefle, Susan L, Aalberse, Rob C, van Ree, Ronald
• Format: Journal Article
• Language: English
• Published: Washington, DC American Chemical Society 15.12.2004
• Subjects: allergens; Biological and medical sciences; Cacao - chemistry; candy; chocolate; Confectionery products and chocolate industries, honey; Corylus - chemistry; Drug Stability; Enzyme-Linked Immunosorbent Assay; food analysis; Food Contamination - analysis; Food industries; food safety; Fundamental and applied biological sciences. Psychology; General aspects; hazelnuts; Methods of analysis, processing and quality control, regulation, standards; pepsin; Pepsin A - metabolism; pepsin resistant allergens; plant proteins; Plant Proteins - analysis; Plant Proteins - metabolism; proteolysis; Bass; Edible fish; Nuts; Enzyme; Pepsin A; Hazelnut; Chocolate; Digestion; Enzyme immunoassay; Chocolates; Protein; food matrix; allergens; Peptidases; Analysis method; pepsin digestion; Hydrolases; Aspartic endopeptidases; Control method; Detection; ELISA assay; Contaminant; Food; ELISA
• ISSN: 0021-8561; 1520-5118
• DOI: 10.1021/jf049278r

Contamination of food products with pepsin resistant allergens is generally believed to be a serious threat to patients with severe food allergy. A sandwich type enzyme-linked immunosorbent assay (ELISA) was developed to measure pepsin resistant hazelnut protein in food products. Capturing and detecting rabbit antibodies were raised against pepsin-digested hazelnut and untreated hazelnut protein, respectively. The assay showed a detection limit of 0.7 ng/mL hazelnut protein or <1 μg hazelnut in 1 g food matrix and a maximum of 0.034% cross-reactivity (peanut). Chocolate samples spiked with 0.5−100 μg hazelnut/g chocolate showed a mean recovery of 97.3%. In 9/12 food products labeled “may contain nuts”, hazelnut was detected between 1.2 and 417 μg hazelnut/g food. It can be concluded that the application of antibodies directed to pepsin-digested food extracts in ELISA can facilitate specific detection of stable proteins that have the highest potential of inducing severe food anaphylaxis. Keywords: ELISA; hazelnut; pepsin digestion; food matrix; allergens