Objectives: Demonstration of the cytotoxic effect of different concentrations of amiodarone (AMD) on HTC cell culture over 24 hrs. The MTT method was used to determine cell viability and the amount of reduced cellular glutathione (GSH) was measured to assess the oxidative status. Microphotography recorded the formation of lipid droplets in treated cells stained with Oil red O (ORO) staining. Study Design: The study was conducted as a randomized controlled trial. Participants and Methods: The research was conducted on hepatoma tissue culture (HTC), a rat liver cell line. Randomization was achieved using a block design and each experiment was repeated three times. Confluent cells were subjected to amiodarone treatment (20 and 40 μM AMD) and after 24 hrs experiments were performed. Three methods were used to determine cytotoxicity: MTT, GSH and ORO staining. Results: The MTT assay showed significantly lower cell survival among treated HTC cells compared to the control group. There is no statistically significant difference between the two groups (20 and 40 μM) treated with AMD. By measuring the amount of GSH, a statistically significant drop in the level of GSH was recorded between the control group and the group treated with 40 μM AMD, while no statistically significant difference was found between the control group and 20 μM, as well as between the treated groups. ORO staining showed a dose-dependent increase in lipid content in treated cells compared to the control group. Conclusion: HTC cells treated with AMD (20 and 40 μM) showed a dose-dependent decrease in cell viability; the occurrence of oxidative stress was recorded by the reduction of GSH when the concentration of AMD increased and insufficient β-oxidation caused the formation of lipid droplets in the cytosol of the treated cells, which was proven by ORO staining.