title: Immunochemical analytical methods for the determination of peanut proteins in foods.
abstract: Peanut proteins can cause allergenic reactions that can result in respiratory and circulatory effects in the body sometimes leading to shock and death. The determination of peanut proteins in foods by analytical methods can reduce the risk of serious reactions in the highly sensitized individual by allowing for the detection of these proteins in a food at various stages of the manufacturing process. The method performance of 4 commercially available enzyme-linked immunosorbent assay (ELISA) kits was evaluated for the detection of peanut proteins in milk chocolate, ice cream, cookies, and breakfast cereals: ELISA-TEK Peanut Protein Assay, now known as "Bio-Kit" for peanut proteins, from ELISA Technologies Inc.; Veratox for Peanut Allergens from Neogen Corp.; RIDASCREEN Peanut Kit from R-Biopharm GmbH; and ProLisa from Canadian Food Technology Ltd. The 4 test kits were evaluated for accuracy (recovery) and precision using known concentrations of peanut or peanut proteins in the 4 food matrixes. Two different techniques, incurred and spiked, were used to prepare samples with 4 known concentrations of peanut protein. Defatted peanut flour was added in the incurred samples, and water-soluble peanut proteins were added in the spiked samples. The incurred levels were 0.0, 10, 20, and 100 microg whole peanut per g food; the spiked levels were 0.0, 5, 10, and 20 microg peanut protein per g food. Performance varied by test kit, protein concentration, and food matrix. The Veratox kit had the best accuracy or lowest percent difference between measured and incurred levels of 15.7% when averaged across all incurred levels and food matrixes. Recoveries associated with the Veratox kit varied from 93 to 115% for all food matrixes except cookies. Recoveries for all kits were about 50% for cookies. The analytical precision, as measured by the variance, increased with an increase in protein concentration. However, the coefficient of variation (CV) was stable across the 4 incurred protein levels and was 7.0% when averaged across the 4 food matrixes and analytical kits. The R-Biopharm test kit had the best precision or a CV of 4.2% when averaged across all incurred levels and food matrixes. Because measured protein values varied by test kit and food matrix, a method was developed to normalize or transform measured protein concentrations to an adjusted protein value that was equal to the known protein concentration. The normalization method adjusts measured protein values to equal the true protein value regardless of the type test kit or type food matrix.
authors.affiliation: U.S. Department of Agriculture, Agricultural Research Service, Box 7625, North Carolina State University, Raleigh, NC 27695-7625, USA. tom_whitaker@ncsu.edu
authors.forename: Thomas B
authors.initials: TB
authors.lastname: Whitaker
authors.forename: Kristina M
authors.initials: KM
authors.lastname: Williams
authors.forename: Mary W
authors.initials: MW
authors.lastname: Trucksess
authors.forename: Andrew B
authors.initials: AB
authors.lastname: Slate
mesh_terms: D000485:Allergens; D010367:Arachis; D002099:Cacao; D002623:Chemistry Techniques, Analytical / Q000379:methods*; D002523:Edible Grain; D004797:Enzyme-Linked Immunosorbent Assay; D005502:Food; D005504:Food Analysis / Q000379:methods*; D007054:Ice Cream; D007120:Immunochemistry / Q000379:methods*; D021183:Peanut Hypersensitivity; D010940:Plant Proteins; D012044:Regression Analysis; D015203:Reproducibility of Results
chemical_list: D000485:Allergens; D010940:Plant Proteins