Erythropoietin (EPO) resistance is an important cause of anemia in patients with chronic kidney disease. Enhancement of erythropoiesis by EPO requires intact EPO signaling and effective mobilization of iron stores. Enhanced synthesis of hepcidin leads to reduction of iron absorption in the small intestine and sequestering of iron in macrophages. Hepcidin may contribute to EPO resistance through a direct inhibitory effect on erythroid progenitor proliferation and survival. Objective To assess serum hepcidin levels in patients undergoing hemodialysis (HD) treatment and to distinguish its analogous association with anemia parameters and EPO resistance. Patients and methods A cross-sectional study was carried out among HD patients at Nephrology Unit of the Zagazig University Hospital from 2017 to 2018. The study included 90 participants, meeting our inclusion criteria, who had end stage renal disease (ESRD) and were on regular HD three times weekly. All patients were subjected to full history taking and clinical and laboratory assessment. It included routine investigations, with complete blood picture, calcium, phosphorus, parathyroid hormone (PTH), lipid profile, iron panel, and c reactive peptide (CRP). Specific investigations included the EPO resistance index and serum hepcidin measurement. Results Hepcidin is positively related to EPO resistance index. EPO resistance index as well as ferritin were significant determinants of hepcidin in HD patients. Conclusion Hepcidin is associated with anemia, iron status, and microinflammation in HD patients. If used as a diagnostic tool, it might improve iron therapy during periods of reticuloendothelial blockage of iron transport. This is important to avoid iron overload and to improve EPO response in patients with ESRD.