Background: Currently, erythroferrone (ERFE) has been discovered to be involved in iron control and erythropoiesis. Objective: To investigate cord blood erythroferrone level and iron status in neonates with high risk factors and correlate these factors to the development of iron deficiency anemia at birth. Patients and methods: This is a prospective study that was conducted on neonates recruited from the Obstetrics and Gynecology Department, Faculty of Medicine, Menoufia University and Neonatal Intensive Care Unit (NICU) during the period from January 2023 to March 2024. The participants underwent full history taking, clinical examination, and laboratory investigations including iron profile and erythroferrone. Results: The mean serum iron and ferritin levels exhibited significant differences among the groups (p < 0.001), with the lowest mean values were shown in the neonates of iron deficient mothers. Total iron binding capacity (TIBC) and cord blood erythroferrone levels showed a statistically significant difference among the groups (p < 0.001), with the highest mean value was shown in the neonates of iron deficient mothers. ROC analysis showed that ERFE had high area under the curves (AUCs) to differentiate neonates of iron-deficient mothers (0.974, p <0.001) and low-birth-weight neonates (0.953, p < 0.001) from the control group. Conclusion: Our findings indicate that neonates of iron-deficient mothers and those with low birth weight exhibit elevated ERFE levels, highlighting the potential of ERFE as a biomarker for diagnosing iron deficiency anemia in high-risk neonates, as demonstrated by the excellent performance of ERFE levels in distinguishing affected neonates from healthy controls. This underscores the clinical relevance of ERFE in neonatal care and its potential as a therapeutic target.