Abstract: Colchicine, a chief alkaloid was determined in six different species of Gloriosa. Solvent extraction of colchicine with petroleum ether and dichlomethane and quantification through High Performance Liquid Chromatography showed high level of colchicine (0.342 mg g -1) in Gloriosa planti amongst the species selected for the study. The mitotic inhibition of colchicine in onion root was standardized using standard colchicine. The effect of colchicine extracted from Gloriosa species was studied in onion root tips treated with 30 mg L -1 colchicine for 2 h. Mitotic abnormalities have been observed and reported for the extracts from different species of Gloriosa under study.