3.9.3. In Vitro α-Glucosidase Assay

The α-glucosidase inhibitory activity was evaluated according to a previously published protocol [19]. The assay mixture consisted of 70 μL of 100 mM phosphate buffer (pH 6.8), 10 µL (0.67 mM) of compound dissolved in DMSO, and 20 µL of 1 U/mL α-glucosidase solution were added to 96 well plates in triplicates. The plate was incubated at 37 °C for 15 min, followed by the addition of 20 μL of the p-nitrophenyl α-D-glucopyranoside substrate. The reaction mixture was incubated at 40 °C for 30 min, and then 50 μL of 0.1 M Na₂CO₃ solution was added. The absorbance was recorded at 405 nm using a microplate reader, and results were expressed in percentage of inhibition. All experiments were conducted in triplicate.