Background. Neonatal jaundice is common a clinical problem worldwide. Globally, every year, about 1.1 million babies develop severe hyperbilirubinemia with or without bilirubin encephalopathy and the vast majority reside in sub-Saharan Africa and South Asia. Strategies and information on determinants of neonatal jaundice in sub-Saharan Africa are limited. So, investigating determinant factors of neonatal jaundice has paramount importance in mitigating jaundice-related neonatal morbidity and mortality. Methodology. Hospital-based unmatched case-control study was conducted by reviewing medical charts of 272 neonates in public general hospitals of the central zone of Tigray, northern Ethiopia. The sample size was calculated using Epi Info version 7.2.2.12, and participants were selected using a simple random sampling technique. One year medical record documents were included in the study. Data were collected through a data extraction format looking on the cards. Data were entered to the EpiData Manager version 4.4.2.1 and exported to SPSS version 20 for analysis. Descriptive and multivariate analysis was performed. Binary logistic regression was used to test the association between independent and dependent variables. Variables at p value less than 0.25 in bivariate analysis were entered to a multivariable analysis to identify the determinant factors of jaundice. The level of significance was declared at p value <0.05. Results. A total of 272 neonatal medical charts were included. Obstetric complication (AOR: 5.77; 95% CI: 1.85-17.98), low birth weight (AOR: 4.27; 95% CI:1.58-11.56), birth asphyxia (AOR: 4.83; 95% CI: 1.617-14.4), RH-incompatibility (AOR: 5.45; 95% CI: 1.58-18.74), breastfeeding (AOR: 6.11; 95% CI: 1.71-21.90) and polycythemia (AOR: 7.32; 95% CI: 2.51-21.311) were the determinants of neonatal jaundice. Conclusion. Obstetric complication, low birth weight, birth asphyxia, RH-incompatibility, breastfeeding, and polycythemia were among the determinants of neonatal jaundice. Hence, early prevention and timely treatment of neonatal jaundice are important since it was a cause of long-term complication and death in neonates.