Aim: This study was designed to assess the impact of sofosbuvir/daclatasvir on vitamin D and iron status in chronic HCV patients (CHC). Methods: Sixty-five CHC patients registered for sofosbuvir/daclatasvir based regimen were recruited. Serum vitamin D, total iron, total iron binding capacity, and serum hepcidin were measured prior the treatment and 12 weeks after the stoppage of the treatment. Transferrin saturation was calculated. Results: The present study showed that a great majority of CHC patients had vitamin D deficiency or insufficiency, high total serum iron, and high transferrin saturation. Vitamin D was negatively correlated ALT. 12 weeks after completion of treatment, patients who had vitamin D deficiency or insufficiency reduced and the median value of vitamin D significantly increased compared to the pretreatment value. Total serum iron, transferrin saturation tended to decrease, however, there were no significant differences. The number of patients who had concurrently vitamin D deficiency or insufficiency and high transferrin saturation was reduced. Serum hepcidin significantly increased 12 weeks after completion of the antiviral therapy. Despite there were no significant correlations between hepcidin and total serum iron, vitamin D, and transferrin saturation before treatment, a significant positive correlation between hepcidin and serum iron and a significant negative correlation between hepcidin and vitamin D were noted after treatment. Conclusion: CHC is associated with vitamin D deficiency and iron overload, which could be attributed to reduced hepcidin level. Treatment with sofosbuvir/daclatasvir increases hepcidin and thereby reduces iron level and its harmful effect on the liver, thus increases vitamin D.