The pharmacological evaluation of substrates from plants is an established method for the identification of compounds, which can lead to the development of novel and safe medicinal agents. The present study supports the previous that brine shrimp bioassay is simple, reliable, and convenient method for assessment of bioactivity of medicinal plants and leads support for their use in traditional medicine. The brine shrimp lethality bioassay is extensively used for the isolation of anitumour and cytotoxic agents from medicinal plants. The present study was designed to assess as well as to compare the cytotoxic activity of Lawsonia inermis Linn. using brine shrimp lethality bioassay. All the extracts of leaves were found to have promising cytotoxic activity. The highest activity was found for leaves which is followed by ethyl acetate and aqueous extract results suggest that the leaves of the Lawsonia inermis Linn. may be a promising source for novel anticancer agents.