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Four hundred (400 ml) of distilled water was added to the separating funnel and the mixture was shaken thoroughly then left to stand until two layers were formed. The bottom layer (organic layer) was collected and the upper layer (aqueous layer) was re-extracted with the chloroform Mitragyna Javanica again and this step was repeated three times. The collected organic layer was filtered through sodium sulphate anhydrous and the organic filtrate was further dried by rotary evaporator. The crude chloroform extract obtained appeared greasy and very dark green in colour.

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