The aim of the study was the evaluation of the cytotoxicity and virucidal activity of the extracts from underground parts of Iris flavissima Pall. The plant material was extracted using: methanol/H₂O (1:1 v/v) and methanol. The examination of the cytotoxicity of the extracts in the concentration of 0.025; 0.05; 0.1; 0.25; 0.5; 1; 2, and 5 mg mL⁻¹ was carried out on GMK cell cultures. The cultures were incubated at 37°C in an atmosphere with 5% of CO₂ for 24-72 h. The cytotoxicity was measured by the colorimetric MTT (tetrazolium) method. Human enterovirus ECHO 9 was used for the evaluation of the virucidal activity of the extracts. The suspension of the virus was mixed (1:1 v/v) with the examined extracts at the concentrations non-toxic to GMK cell cultures and incubated at 37°C for 1 h, and then the virus was titrated in the GMK cell culture. The titre of the virus was estimated according to the Reed-Muench method. The methanolic and methanol/H₂O extracts were found to be non-toxic to GMK cells at the concentration of 1 mg ml⁻¹ and 2 mg ml⁻¹, respectively. The virucidal effect was observed for the both investigated extracts. The methanolic extract decreased ECHO 9 replication by 1.46 log, and methanol/H₂O extract - by 2.08 log.