**4. Extraction and fractionation procedure**

Extraction and fractionation were according to Gandhi et al. [28] and Leila et al. [29] with some modification in the choice of primary solvent (water) and partitioning solvents (hexane, chloroform, ethyl acetate, and butanol) [26]. The solvents are chosen in order of polarity. The advantage is to allow for different chemical compounds in the plant part (seed) to selectively dissolve in solvents they are accommodated in (as with the chemical axiom "like dissolves like"). The extracted compound is collected from the rotary evaporator under vacuum at 45°C and is ready for use.

 The ethanol extract residue obtained is dissolved in 100% water (500 ml) and exhaustively extracted by consecutive liquid/liquid partition with hexane (500 ml), chloroform (500 ml), ethyl acetate (500 ml), and saturated butanol (500 ml) using a separating funnel (1000 ml). That is, on your initial fill of the separating funnel with primary solvent (500 ml water) and the extracted compound, shake vigorously to make a solution before adding the next solvent. The content is again shaken vigorously following the addition of the next solvent, before it is left to stand till the obvious separation of both solvents. The separated solvent is let out into a container and labeled the solvent (say hexane) extract. The next solvent (500 ml) of choice (chloroform) is again added to the remaining water extract solution left in the separation funnel and again shaken vigorously. The higher the number of solvent used is, the better the expected result and further ease at subsequent separation. The experiment takes advantages of the immiscibility of the solvents with water (the primary solvent used to dissolve the extracted compound).

The hexane, chloroform, ethyl acetate, saturated butanol, and last remaining aqueous fractions are evaporated to obtain fractions [26]. It should be borne in mind that each of the fractions contains a portion of some of the chemical constituent in the primary water-extract mixture in the initial fill of the funnel. The fractions obtained (hexane, chloroform, ethyl acetate, saturated butanol, and last remaining aqueous) are bioassayed for bioactivity. At this stage, the entire model met for sufficient scientific authentication of the disease (investigated) is used as with the case of the crude extract [25]. That is tested for antidiabetic and phytochemical properties in the case of this writer/researcher.
