The methanolic extract
0.5 mL was added to 0.5 mL Foline Ciocalteu reagent (1:10) in test
tubes and manually shaken for 15–20 s. The reaction mixture was
added to 1 mL of 7.5% sodium carbonate solution and 1 mL of
distilled water. The reaction mixture was incubated in the dark for
2 h at room temperature and the absorbance was read at 725 nm.
The total phenolic content was calculated from calibration curve
using gallic acid as a standard and then expressed as mass of gallic
acid equivalents (GAE) per mass of tissue on a fresh weight basis
(g kg1).