1.3.4. Fractionation
The hydro ethanolic crude extract (190g) was fractioned in hexane, ethyl
acetate, n-butanol in increasing order of polarity. The hydro ethanolic
extract was initially mixed with 500 ml of distilled water and 450 ml of
hexane then transferred into a separating funnel. After shaking and
decantation, the separation of two phases was obtained. The upper
organic phase which represents the hexane fraction was transferred in to
an erlenmeyer. The lower organic phase was extracted many times by
adding fresh solvent (hexane) until a clear phase was obtained. The
hexane fractions was then pooled and stored as such. The lower phase was
mixed with subsequent solvents (ethyl acetate and then n-butanol) and
they followed same treatment as hexane fraction. At the end of the
fractionation the residue was called residual fraction. The four
fractions were concentrated and dried.