*2.4. Total Phenolic Content*

The colorimetric reaction with FCR was carried out to determinate the total phenolic content (TPC) [27]. The 0.25 mL of extract solution in methanol (1.25 mg/mL), 0.25 mL of FCR, 0.5 mL of saturated sodium carbonate solution, and 4 mL of water were mixed. After 25 min, the reaction mixtures were centrifuged (5 min, 5000× *g*, MPW-350R, MPW Med. Instruments, Warsaw, Poland), and the absorbance of supernatants was measured at 725 nm (DU-7500 spectrophotometer, Beckman Instruments, Fullerton, CA, USA). TPC was expressed as gallic acid equivalents (GAE) per g of extract or per g of FM of plant.
