Plumbago zeylanica is a species of the genus Plumbago in the family Plumbaginaceae. Different parts of this plant are used in traditional practice of medicine. The objective of the study was to analyses the phytochemical constituents of P. zeylanica root and evaluation of anticancer activity. Three different extracts petroleum ether, chloroform and methanolic extracts of P. zeylanica roots were prepared and phytochemical screenings were carried out. In the extracts, compounds like alkaloids, flavonoids, phenolic compounds, saponins, tannins and terpenoids are detected by phytochemical analysis. Based on the results of phytochemical assays, methanolic extract was selected for evaluation of its anticancer activity using HeLa and A375 cell lines. The present study showed that P. zeylanica root contains bioactive compounds that are known to possess anticancer activity and the methanolic extract caused decline in cell viability as the concentration of extract increased.