Bioassay Guided Chromatographic Isolation Of Lactation Inducing Agent From Hippocratea Obtusifolia

Abstract

The powdered leaves of Hippocratea obtusifolia (commonly known as gwadayi in Hausa) was percolated with ethanol and then sequentially extracted with petroleum ether, chloroform, ethyl acetate and methanol. The extracts were respectively labeled HO1, HO2, HO3, and HO4, with the ethanolic extract labeled HO. The extracts were subjected to bioactivity against lactation inducement, and the chloroform extract labeled HO2 was found to be the most active. The HO2 extract was then subjected to column and thin layer chromatography with similar fractions pooled together. The pooled fractions were then subjected to bioactivity testing against lactation inducement, and the fraction labeled HO2-13 was found to be the most active, this was subsequently subjected to column and thin layer chromatography with similar fractions pooled together. The pooled fractions were again subjected to bioactivity testing against lactation inducement, and the fraction labeled HO2-13-111 was found to be the most active. The fraction HO2-13-111 was subjected spectroscopic analysis, using Infrared, Mass spectroscopy, and both ¹H and ¹³C NMR data obtained. The structure of the compound present in fraction HO2-13-111 was then elucidated based on the obtained data.

Keywords: Force-Feeding, Lactation, Progesterone, Thin-Layer Column Chromatography.