This study was aimed to investigate the iron overload effect on the levels of ferritin, testosterone, follicle stimulating hormone (FSH), luteinizing hormone (LH) and prolactin (PRL) in male patients with β-thalassemia. Blood levels of sex hormones and ferritin were determined in (50 males) beta-thalassemia patients, aged (16-23) years and in (30) healthy males matched for age. They were recruited from the Abin Al-Balady Hospital in Baghdad. Ichroma™ kits were used to determine the blood levels of sex hormones and ferritin. The results showed that the level of testosterone, LH and FSH were significantly (p≤ 0.05) lower in β-thalassemia male patients compared to controls. Furthermore, the level of ferritin and prolactin in male patients were significantly (p≤ 0.05) higher than the values obtained in the controls. Significant positive correlations (p≤ 0.05), (r= 0.3834) were detected between level of LH and ferritin in male patients. Non-significant positive correlations (p≥0.05) in the level of testosterone and ferritin, also non-significant negative correlation in PRL and FSH and ferritin were found in patients with β-thalassemia. From the above results it can be concluded that the iron overload may be the major cause of infertility in β-thalassemia male patients.