A colorimetric method for screening α-glucosidase inhibitors from flavonoids using 3,3′,5,5′-tetramethylbenzidine as a chromogenic probe

A facile and novel colorimetric method for screening of α-glucosidase inhibitors (AGIs) from flavonoids using 3,3’,5,5’-tetramethylbenzidine (TMB) as a chromogenic probe is proposed. This method is based on the colorimetric detection of ascorbic acid (AA) through the TMB oxidation reaction catalyzed by horseradish peroxidase (HRP) in the presence of hydrogen peroxide (H2O2). In the TMB/H2O2/HRP system, HRP catalyzes the oxidation of H2O2 to ‧OH radical which oxidizes TMB to blue-colored oxidized TMB (oxTMB). In the presence of AA, the production of ‧OH radical is suppressed and causes the decrease of oxTMB, resulting in the fading of the blue color and the decrease of absorbance at 652 nm. Based on this, the existence of AA can be facilely identified. In the 2-O-α-d-glucopyranosyl-l-ascorbic acid (AA-2 G)/α-glucosidase (α-Glu) system, the produced AA inhibits the oxidation of TMB to blue-colored oxTMB. In the presence of AGIs, the production of AA is inhibited, which inhibits the reduction of oxTMB, resulting in a blue color recovery and an increase of the absorbance at 652 nm. Based on this, the colorimetric method is developed for screening of AGIs from 7 flavonoids.