Rapid analysis of wheat gluten composition using a triple ELISA

Abstract

AbstractBACKGROUNDGluten composition is an important quality parameter of wheat flour. Reversed‐phase high‐performance liquid chromatography (RP‐HPLC) is a state‐of‐the‐art method for its analysis. As this is a very labour‐intensive and time‐consuming procedure, alternative faster methods are desirable. Enzyme‐linked immunosorbent assay (ELISA) is a high‐throughput method often used for the analysis of gluten traces in gluten‐free products. In this proof‐of‐principle study, we introduce an experimental triple ELISA for the relative quantitation of gliadins, high‐molecular‐weight glutenin subunits (HMW‐GS) and low‐molecular‐weight glutenin subunits (LMW‐GS) of one wheat flour extract.RESULTSThe results of 80 common wheat flour samples obtained from the triple ELISA and RP‐HPLC were correlated. The results for gliadins (r = 0.69) and HMW‐GS (r = 0.81) showed a medium and high correlation, respectively. Only a very weak correlation of ELISA and RP‐HPLC results was observed for LMW‐GS (r = 0.49). Results for glutenins (r = 0.69) and gluten (r = 0.72) had a medium correlation. The gliadin/glutenin ratio (r = 0.47) and LMW‐GS/HMW‐GS ratio (r = 0.40) showed a weak or no correlation. The gliadin, LMW‐GS and gluten contents were lower and the HMW‐GS content was higher in the ELISA measurement compared to RP‐HPLC.CONCLUSIONThe quantitation of gliadins and HMW‐GS by the experimental triple ELISA showed comparable results to RP‐HPLC, whereas no strong correlation between the results from the two methods was found for LMW‐GS. Overall, the experimental triple ELISA is suitable for relative gluten quantitation, especially for the analysis of large sample sets. Further work will focus on improving the experimental procedure of the ELISA. © 2024 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.