The presence of Colchicine in Gloriosa superba was confirmed by Thin Layer Chromatography (TLC) and High Pressure Liquid Chromatography (HPLC) in which a spot coinciding with the authentic sample of Cochicine in Rf value (Colchicine 0.70) appeared. The maximum amount of Colchicine was found in flower (20.7 mg/gdw) and minimum amount was found in stem (7.4 mg/gdw).The In vitro studies showed the maximum amount of Colchicine in 6 week old cultures (22.6 mg/gdw)) and minimum amount in 2 weeks old cultures (15.3 mg/gdw). Key words: Colchicine, Gloriosa superba, HPLC Laboratory of Bioactive Natural Products, Department of Botany, University of Rajasthan, Jaipur -302004 * Corresponding Author, Email: [email protected]; Tel: +91-141-2701875, +91-9828070584 Please Cite This Article As:Renu Sarin, Anirudha Rishi and Ajit Kumar. 2010. In vivo and In vitro Estimation of Colchicine in Gloriosa superba L. by High Pressure Liquid Chromatography. J. Exp. Sci. 1(4):1-2.Â