2.5.2. Total Flavonoids Content (TFC)

Total flavonoid content for all sample extracts was measured by the AlCl₃ colorimetric method [22] with slight modifications using a 96-well plate. 80 µL sample extract was mixed with 80 µL of 2% AlCl₃ solution. 120 µL of aqueous solution of sodium acetate (50 g/L) was added. This mixture was incubated for 150 min at 25 °C. Absorbance was taken at 440 nm. Measurements for all samples were made in triplicate, and quantification was done by constructing a standard curve (0–50 µg/mL quercetin).