2.8. Total Flavonoids Assay

Total flavonoid content was measured by AlCl₃ colorimetric assay, executed by [29]. Briefly, 1 mg/mL of plant extract/standard solutions of quercetin (20, 40, 60 and 100 μg/mL) was added to a volumetric flask having 4 mL of dH₂O. In addition, 0.30 mL 5% NaNO₂ and after five minutes 0.3 mL 10% AlCl3 were added in a flask, respectively. Then, IM NaOH (2 mL) was added, and the volume was prepared up to 10 mL with distilled water. Absorbance was taken against the blank at 510 nm. TFC was articulated as mg Quercetin Equivalents (mg QE).