Investigation of different extraction conditions on the efficiency of gliadin extraction and determination by ELISA method

Abstract

Gluten refers to a complex mixture of gliadins and glutenins. It can cause numerous foodborne disorders. In sensitive individuals gluten can lead to celiac disease (CD), wheat sensitivity and allergy. Gliadin proteins are one of the gluten fractions. The aim of this paper was to examine how different conditions, mixing time (2.5, 5, 7.5, and 10 min) of the sample with the most commonly solvent 70% (v/v) ethanol and 70% (v/v) isopropanol and incubation time (15, 20, 25, and 30 min) affect the efficiency of gliadin determination with the ELISA method. A commercial kit was used to determine gliadin concentrations, and absorbance was measured at 450 nm, using the ELISA reader. Based on the obtained results, the optimal mixing time of the sample with the solvents was 5 minutes and the incubation time was 25 minutes. Under these conditions, the extraction efficiency is the best, i.e., the highest gliadin concentration is obtained. The results of research can be of fundamental importance in the study of gluten proteins and the impact of technological procedures on their change and the possibility of reducing the allergic effect.