Iron overload which results from repeated blood transfusion is a serious problem as it may affect many organs as liver causing fibrosis and then progress to cirrhosis and pituitary gland leading to pituitary hypofunction which may affect sexual maturation. So, iron overload must be detected accurately and quickly. The ordinary methods for detection of iron overload as measurement of serum iron, transferrin saturation, TIBC, and serum ferritin are not accurate methods as they are affected by many diseases and conditions such as hepatitis, fever, chronic inflammation, and hemolysis and ascorbate deficiency. Non- transferrin bound iron is an accurate method for diagnosis of iron overload as it is present only in serum of patients with iron overload and not present in serum of normal individuals or patients with other diseases.