Abstract: Background: Ulcerative Colitis (UC) is a subcategory of inflammatory bowel disease and it refers to a large group of disorders that affect the gastrointestinal system. It has many serious effects and large population from European country is suffering from this disease so attempts to find new medicines from natural sources are needed. Objectives: The roots of Tephrosia purpurea (Leguminosae) are used in the treatment of ulcer and colic pain traditionally hence present work was undertaken to validate it scientifically and to identify the phytoconstituent responsible for this activity. Material and Methods: Various extracts of root were screened for Ulcerative Colitis (UC) using the method acetic acid induced ulcerative colitis in mice. Aqueous extract was found most active and hence fractionated by column chromatography. Various fractions obtained were screened for UC activity and fraction IV was found most active hence purified and subjected to spectroscopic analysis yielding a glycoside. Results: Results showed that fraction IV is found to contain a glycoside after spectroscopic study. This glycoside reduced level of myeloperoxidase and malondialdehyde in colon significantly. This glycoside showed significant protective effect in macroscopical study and histopathology observations of the colon. Conclusion: It can be concluded that the glycoside, 1, 4a, 7, 7a-tetrahydro-7-hydroxy-7-(hydroxymethyl)-1-(tetrahydro-3’,4’,5’-trihydroxy-6’-(hydroxymethyl)-2H-pyran-2’-yl oxy) cyclopenta (c) pyran-4-carboxylic acid might be responsible for the treatment of ulcerative colitis.