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In the present study we used the micronuclei test and the comet assay in mice to investigate the genotoxic and mutagenic effects of three benzopyrans--6-isobutyryl-5,7-dimethoxy-2,2-dimethyl-benzopyran (HP1); 7-hydroxy-6-isobutyryl-5-methoxy-2,2-dimethyl-benzopyran (HP2); and 5-hydroxy-6-isobutyryl-7-methoxy-2,2-dimethyl-benzopyran (HP3)--isolated from Hypericum polyanthemum. No significant difference in polychromatic erythrocyte (PCE) percentage between the vehicle group and groups treated with HP1, HP2, or HP3 was detected, indicating no toxicity to the bone marrow of the animals. Only HP1 increased the frequency of micronucleated cells (MNPCEs) in bone marrow sampled after 24 h in comparison with the vehicle group, suggesting a weak mutagenic effect. The damage index and damage frequency did not show a significant increase after treatment with HP1, HP2, or HP3 in comparison with the vehicle group. The antitumor activity previously reported in vitro for these benzopyrans, the lack of acute toxicity, the MN induction only for HP1, and the relatively low DNA damage make all compounds good candidates for in vivo studies on antitumor action.