For the investigation of bioactive natural products with hypoglycemic effect, we have evaluated various extracts of Pulsatillae Radix(Ranunculaceae), which has been used in traditional medicine for hematochezia due to intense evil heat, malaria, chills and fever, epistaxis and internal hemorrhoids. The ethylacetate extract of the radix of this plant was found to show a significant hypoglycemic effect on alloxan diabetogenic mice. Using bioactivity-guided chromatographic purification of the ethylacetate extract, hypoglycemic constituent: 2${eta}$, 3${eta}$, 14${alpha}$, 20, 22R, 25-hexahydroxy-cholest-7-en-6-one was isolated and structurally identified by physico-chemical properties and spectroscopic evidences.