The method described by51 was used with modification. One gram of the sample was soaked in 20 ml of 20% ethanol, heated in a water bath at 55 °C for 4 h, and stirred well. The content was filtered and re-extracted with 20 ml of 20% ethanol and shaken vigorously (not heated). This was filtered and the filtrate was added with the first filtrate. The filtrate was then reduced to one-quarter of its value in the water bath at 90 °C. This content was later poured into a separating funnel and 20 ml of diethyl ether was added into it. The content was shaken vigorously and attach back to the tripod stand. Aqueous / lower layer of the content was collected and ether/ upper portion containing fatty substances was discarded. The aqueous layer was then poured back into the funnel, and n- butanol was added and then shaken vigorously. A clean pre-weighed beaker was used to collect the butanol layer in the weighed beakers and evaporated to dryness. The residue was later dried in an oven until a constant weight is obtained.

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