a hydro-alcoholic mixture (methanol / water 80/20: v/v) for
72 hours, in the dark at room temperature, with renewal
of solvent every 24 hours (350 ml x 3). Macerate was
filtered through filter paper to obtain a clear extract, and
then solvent was evaporated under vacuum at a
temperature of 40° C, 200 ml of distilled water was
added to the residue, extract was kept for 48 h in a
refrigerator, then decanted, and was filtered again
through filter paper Lim et al. (2012).
The liquid-liquid extraction has been then carried out
with three organic solvent system of increasing polarity:
200 ml of petroleum ether, 200 ml of chloroform, 200 ml
of ethyl acetate and finally 200 ml of 1-butanol
Benhammou et al. (2007). The residue of each solvent
extract was dissolved in water, frozen and lyophilized.
Chloroform, ethyl acetate, 1-butanol and aqueous crude
extract powder were used for investigate phytochemical
compounds, determination of total phenols, flavonoids
content, antioxidant screening and for HPLC.