Iron deficiency (ID) in premature infants is known to be associated with a delay in neurodevelopmental outcomes. Therefore, timely diagnosis of this condition is extremely important. Hepcidin (Hep) is a hormone regulates iron metabolism and can be used as an additional marker for the diagnosis of ID. The objective of the study was to evaluate the correlation of serum Hep concentration with erythrocyte’s indexes (MCV, MCH, MCHC, RET-He) and iron metabolism indicators (serum ferritin, iron, transferrin), and to determine the diagnostic significance of Hep for the diagnosis of ID in premature infants weighing less than 1000 g at birth in the late neonatal period. Material and methods. The prospective cohort study included 106 infants weighing less than 1000 g at birth. Infants at the age of 14 to 21 days of life were laboratory examined including determination of MCV, MCH, MCHC, RET-He, serum Hep, serum ferritin, serum transferrin and serum iron. Depending on the presence of ID, infants were divided into 2 groups: group 1 – infants with ID (n=55); group 2 – infants without ID (n=51). Results. The significant correlations of serum Hep with such indicators as serum ferritin (r=0.97, p=0.012), MCV (r=0.71, p=0.020), MCH (r=0.62, p=0.035), MCHC (r=0.61, p=0.035), RET-He (r=0.77, p=0.023) were determined. Serum Hep was significantly lower in infants with ID. The threshold value of Hep (≤6.4 ng/ml) was determined, associated with a high probability of ID (Se=86.7% and Sp=76.4%). A low serum Hep indicates a lack of total iron storage in premature infants, reflecting an increased need for iron in this category of infants. The difference between serum iron and transferrin concentrations was not statistically significant among the groups. Conclusion. The correlations between serum Hep, ferritin, MCV, MCH, MCHC, RET-He were found in premature infants weighing less than 1000 g at birth in the late neonatal period. The diagnostic value of determining the concentration of serum Hep in the presence of ID has been demonstrated. © 2024 Geotar Media Publishing Group. All rights reserved.