ABSTRACT Background: β thalassemia patients have repeated blood transfusions those lead to iron overload, resulting in iron toxicity. In such cases Hepcidin results in excessive iron absorption. Iron accumulation in organs leads to several complications. Erythroferrone (ERFE) is a hormone that regulates iron metabolism through its actions on hepcidin. Objective: we aimed to study the role of erythroferrone and its relationship with hepcidin in B-thalassemia patients (major –intermedia). Methodology: This Prospective case control study was conducted on 50 participants. 13 β thalassemia major (β- TM) patients (group Ia) and 12 β thalassemia intermedia (β- TI) patients (group Ib). In addition to, 25 healthy individuals included as control group (group II). Routine laboratory investigation, serum hepcidin, serum erythroferrone and iron markers were measured in all groups. Results: In this study, there was significant increase in erythroferrone in group Ia and group Ib while hepcidin showed significant decrease in group Ia and group Ib when compared to a control group (group II) with P value <0.001 , There was significant increase in serum ferritin, total iron binding capacity (TIBC), Serum iron and transferrin saturation (TS) in group Ia and group Ib when compared to a control group (group II). There was negative significant correlation between hepcidin and RDW, significant positive correlation with MCH, MCV. There was negative significant correlation between ERFE and MCV, MCH. Conclusions: Expression of ERFE is altered in multi transfused β thalassemia patients. Extended studies should be performed to assess its discriminative ability between β thalassemia major and minor.