Iron deficiency anemia (IDA) is the one of the most common nutritional problems and is encountered all over the world. This study analysed the effects of squid ink melanin-Fe (SM-Fe) on IDA in rats. Forty weanling SD male rats were used and thirty-two rats were fed an iron-deficient diet for 4 weeks. Then SM-Fe (dosages of iron is 6 mg kg(-1) BW) was given to the IDA rats once a day for 3 weeks by intragastric administration, with FeCl3 and FeSO4 (dosages of iron is 6 mg kg(-1) BW) as positive controls. While the IDA model group and the control group were administrated distilled deionized water each day for 3 weeks. The content of haemoglobin (Hb), serum iron (SI), total iron binding capacity (TIBC), serum ferritin (SF), transferrin receptor (sTfR), erythropoietin (EPO), and iron content in the liver and spleen were measured. The results showed that the content of Hb, SI, SF, EPO, iron content in the liver and spleen were significantly increased in the iron supplement groups (SM-Fe, FeCl3 and FeSO4) compared with the model group (P < 0.05), while TIBC and sTfR were significantly decreased in the iron supplement groups compared with the model group (P < 0.05). In comparison with the FeCl3 and FeSO4 groups, a higher bioavailability of iron and fewer side effects were observed in the SM-Fe group. The present study indicated that SM-Fe is an effective source of iron supplement for IDA rats and might be exploited as a new iron fortifier.