Ethnopharmacological relevance Dryopteris crassirhizoma has been traditionally used for the treatment of tapeworm infestation, the common cold and cancer in Korea, China and Japan. Despite various pharmacological properties of Dryopteris crassirhizoma, there is no available information about the safety of Dryopteris crassirhizoma. Aim of this study To ensure more information about the safety of Dryopteris crassirhizoma, we performed the acute oral toxicity and genotoxicity tests of Dryopteris crassirhizoma. Materials and methods The acute oral toxicity test of Dryopteris crassirhizoma was performed in rats. Genotoxicity of Dryopteris crassirhizoma was evaluated by bacterial reverse mutation, chromosomal aberration and bone marrow micronucleus tests. Results In acute toxicity test, Dryopteris crassirhizoma exhibited no mortality, body weight and behavioral changes and adverse effects in male and female rats. Dryopteris crassirhizoma did not significantly increase the number of the bacterial revertant and chromosomal aberration in both in vitro assays. Moreover, the Dryopteris crassirhizoma-related increases of micronucleated polychromatic erythrocytes (MNPCE) in mouse bone marrow were not observed. Conclusion Therefore, Dryopteris crassirhizoma is non-genotoxic in a three standard battery of tests and the oral LD50 of Dryopteris crassirhizoma is >2000mg/kg.