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The essence of the present study is to focus on the cytotoxicity of the aqueous and solvent (acetone, methanol and ethanol) of the medicinal plant (Calotropis procera) belonging to the family Apocynaceae. Brine shrimp lethality bioassay method was established for the present study and the cytotoxicity was reported in terms of lethality concentration (LC 50 ). The shrimps were hatched and active shrimps were collected and used for the assay. Ten active naupilli were added to the 1ml of the diluted extracts, concentration was varied from 10, 100, 1000µg/ml. The surviving (larvae) shrimps were counted after 24hrs, 48hrs and lethality concentration LC 50 was also assessed. It was observed that the methanol extract of Calotropis procera leaf have weak cytotoxic compounds while other extracts were able to cause 45% mortality of brine shrimp larvae after 48hours. The present study supports that brine shrimp bioassay is simple, reliable and convenient method for assessment of bioactivity and cytotoxicity of medicinal plants. The cytotoxicity exhibited by the crude extracts were promising and this indicates the presence of potent bioactive compounds.