Objective and backgroundPerinatal asphyxia is a serious medical condition in which there is a lack of oxygen or blood flow during, or even after the labor process. So this study aimed to study serum blood levels of Lactate, vitamin D and count of Nucleated Red Blood Cells (NRBCs) per 100 White Blood Cells (WBCs) as early biological predictors of Hypoxic–Ischemic Encephalopathy (HIE) after birth Asphyxia. MethodsA cross-sectional case-control study was conducted on 20 neonates with HIE comprising the cases (APGAR ≤ 5) and another 20 neonates representing the control group with their age, sex, and weight matched, admitted to NICU from March 2020 to the end of September 2020. Venous blood samples for lactate, vitamin D and nucleated RBCs were drawn after birth and sent for analysis. Sarnat score was used to assess the severity of HIE. ResultsVenous blood levels of lactate and nucleated red blood cells per 100 white blood cells (NRBC/100 WBC) counts were significantly higher in HIE neonates than in the control group with a highly significant difference (P-value <0.001). While serum Vitamin D was significantly lower in the HIE group than in the control (P-value <0.001). ConclusionSerum lactate level, vitamin D level and NRBCs/100 WBCs might be used as biomarkers in the early prediction of HIE.