In this study, we aimed to investigate the protective effect of Vitamin U (Vit U) on amiodarone (AMD)-induced hepatotoxicity. Male Sprague-Dawley rats were randomly divided into four groups. Group I was control animals receiving corn oil for 7 days, Group II consisted of animals receiving Vit U (50 mg/kg) for 7 days, Group III consisted of animals receiving AMD (100 mg/kg) for 7 days and Group IV consisted of animals given AMD and Vit U (in same dose and time). AMD and Vit U were administered to rats by gavage. On the 8th day, all the animals fasted overnight were sacrificed. Blood and liver tissue samples were taken from animals. Serum total cholesterol levels and liver gamma glutamyl transferase (transpeptidase) adenosine deaminase and tissue factor activities, liver glycoprotein component level such as fucose, hexose and hexosamine were determined. All the parameters were found to be increased in AMD group as compared to control group. Administration of Vit U reversed these effects in AMD group. According to these results, we can conclude that Vit U can prevent AMD-induced liver injury.