Introduction: Anemia is an abnormal decrease in red blood cells (RBCs) and is classified as; macrocytic, normocytic or microcytic. Iron profiling along with complete blood counting (CBC) is performed to diagnose microcytic hypochromic anemia (MCA). Aims: To evaluate the role of iron profile in differentiating MHA. Methodology: A retrospective chart-review study was conducted in 2019 on anemic patients attending King Abdulaziz Medical City. Values from CBC and iron profiling tests were collected. Results: 219 patients with MHA, 164 females and 55 males, were identified. Female patients were 74.9% of all cases, and adults were the most affected. Values of CBC parameters including hemoglobin, RBCs, packed cells volume (hematocrit), mean cell volume and mean cell hemoglobin were significantly lower in cases than in controls (p__ampersandsignlt;0.001). Iron profiling showed a significant association between all parameters and the diagnosis of iron deficiency anemia (IDA) and anemia of chronic disease, but not with TIBC. Iron profile values in thalassemia showed a significant association between diagnosis and serum iron and ferritin only. In sideroblastic anemia, iron profile values showed a significant association between all parameters except for transferrin. Conclusion: IDA and anemia from chronic diseases are the highest among MHA. Iron profiling plays an important role in differentiating MHA.