Chebulinic acid showed many bioactivities including inhibition of cancer cell growth, inhibiting the contractile responses of cardiovascular muscles, anti-fungal, anti-bacterial activities etc. In-vitro anti-cancer activity of Chebulinic acid on Colon adenocarcinoma HT-29 cancer cell lines by using MTT cell growth inhibition assay was stuied. The maximum percentage inhibition of cancer cell lines for Chebulinic acid was found to be 41.2% at a dose of 200µg/ml. Chebulinic acid extraction from Terminalia chebula plays a vital role in medicine, biotechnology and various pharmacological activities.