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A novel approach for the detection of potentially hazardous pepsin stable hazelnut proteins as contaminants in chocolate-based food.

Akkerdaas, J.H., M. Wensing, A.C. Knulst,O. Stephan, S.L. Hefle, R.C. Aalberse,R. van Ree, 2004. A novel approach for the detection of potentially hazardous pepsin stable hazelnut proteins as contaminants in chocolate-based food. J Agric Food Chem. 52:7726-7731.

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.