The total phenolic content (TPC) in each fracton was determined by the Folin–Ciocalteu method . One milliliter of extract in methanol was mixed with 1 ml of 3-fold-diluted Folin–Ciocalteu phenol reagent. Two milliliters of 35% sodium carbonate solution and distilled water were added subsequently. The mixture was incubated for 30 min and the absorbance was recorded at 700 nm by a spectrophotometer