2.7.1. Anti-Tyrosinase Assay

The tyrosinase inhibitory activity of extracts was evaluated using a dopachrome method with L-3,4-dihydroxyphenylalanine (L-DOPA) as the substrate. Briefly, the samples were diluted at 1 mg/mL in 0.1 M potassium phosphate buffer (pH 6.8). Forty microliters of extract, 80 µL of buffer, and 40 µL of mushroom tyrosinase (100 units/mL) (dissolved in buffer) were mixed in 96-well microplates and incubated for 10 min at room temperature. Then, 40 µL of 10 mM L-DOPA (dissolved in buffer) was added. After a 10 min incubation, the absorbance was measured at 475 nm using an absorbance microplate reader. Kojic acid, which inhibits tyrosinase, was used as the positive control (IC₅₀ = 7.11 µg/mL).