Isolation and characterization of glucocorticoid steroid was carried out on the leaf of Rauvolfia vomitoria. Initial phytochemical screening of the sample revealed the presence of saponins, flavonoids, alkaloids, tannins, steroids, triterpenoids and cardiac glycoside and were quantitatively determined as follows; saponins (2.37 ± 1.09), tannins (0.09 ± 0.01), alkaloids (0.82 ± 1.05), and flavonoids (0.82 ± 0.05). Isolation of the compound was carried out with the aid of column chromatography and isolate subjected to thin layer chromatography. Sample was subjected to spectroscopic analysis with the aid of FT-IR, 1H NMR, 13C NMR and HSQC. The structure of compound was determined as Cis-3-(2,2-dimethyl-26-(((2S,3R,4S,5R,6R)-2,3,4-trihydroxy-5-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)butan-2-yl)-8-methoxy-15-(S)-3-methoxy-4,4- dimethylpentyl)-14-(3-methoxy-4,4-dimethylpent-1-yn-1-yl)-1,2,3,4,11,12,13,14,15,15a- octahydrophenanthro [2,1-b]furan-16(10H)-one with a molecular formulae of C44H68O11. A glucocorticoid steroid.