Effect of Processing on
Recovery and Variability Associated
with Immunochemical Analytical Methods for Multiple Allergens in a
Single Matrix: Dark Chocolate
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Abstract
Immunodetection of allergens in dark chocolate is complicated
by
interference from the chocolate components. The objectives of this
study were to establish reference materials for detecting multiple
allergens in dark chocolate and to determine the accuracy and precision
of allergen detection by enzyme-linked immunosorbent assay (ELISA)
before and after chocolate processing. Defatted peanut flour, whole
egg powder, and spray-dried milk were added to melted chocolate at
seven incurred levels and tempered for 4 h. Allergen concentrations
were measured using commercial ELISA kits. Tempering decreased the
detection of casein and β-lactoglobulin (BLG), but had no significant
effect on the detection of peanut and egg. Total coefficients of variation
were higher in tempered than untempered chocolate for casein and BLG,
but total and analytical CVs were comparable for peanut and egg. These
findings indicate that processing has a greater effect on recovery
and variability of casein and BLG than peanut and egg detection in
a dark chocolate matrix