Medicinal plants constitute an important component of flora and are widely distributed in India. The pharmacological evaluation of substances from plants is an established method for the identification of lead compounds which can leads to the development of novel and safe medicinal agents. Based on the ethnopharmacological literature, several species of medicinal plants used in traditional medicine in India were collected. In the present study, aqueous extracts of these medicinal plants were screened for their cytotoxicity using brine shrimp lethality test. Out of the 120 plants tested, Pistacia lentiscus exhibited potent brine shrimp lethality with LC50 2.5 μg. Aristolochia indica (Aristolochiaceae), Boswellia serrata (Burseraceae), Ginkgo biloba (Ginkgoaceae), Garcinia cambogia (Clusiaceae), and Semecarpus anacardium (Anacardiaceae) have also showed significant cytotoxicity with LC50 13, 18, 21, 22, and 29.5 μg respectively. The present study supports that brine shrimp bioassay is simple reliable and convenient method for assessment of bioactivity of medicinal plants and lends support for their use in traditional medicine.