Background: Pre-lacteal feeding is associated with infant morbidity and mortality especially during the neonatal period. About 96% infant deaths in developing countries are attributable to inappropriate feeding practice during the first 6 months of life. This study assessed determinants of pre-lacteal feeding practices in Ethiopia using the data from nationally representative survey. Methods: Data were extracted from the 2016 Ethiopian Demographic and Health Survey to assess determinants of pre-lacteal feeding practices in Ethiopia. The analysis included a weighted sample of 5303 mothers having children aged 0–36 months. A multivariable logistic regression analysis was conducted and the results were presented with adjusted odds ratio at 95% confidence interval, declaring statistical significance at a p-value < 0.05 in all analyses. Results: From a total of 5303 mothers having children aged 0–36 months, 423 (8%, 95% confidence interval, 7.06%, 8.99%) had given pre-lacteal foods to their newborn baby. Being from agrarian region (adjusted odds ratio = 0.15, 95% confidence interval, 0.11, 0.20), poorest wealth status (adjusted odds ratio = 1.50, 95% confidence interval, 1.02, 2.22), home delivery (adjusted odds ratio = 1.35, 95% confidence interval, 1.01, 1.79), late initiation of breast feeding (adjusted odds ratio = 4.52, 95% confidence interval, 3.62, 5.64), having no counseling on breast feeding (adjusted odds ratio = 1.33, 95% confidence interval, 1.01, 1.75), and cesarean delivery (adjusted odds ratio = 2.47, 95% confidence interval, 1.45, 4.20) were factors significantly associated with pre-lacteal feeding practice. Conclusion: A significant proportion of Ethiopian mothers had given pre-lacteal foods to their newborn babies. Poorest wealth index, region, late initiation of breast feeding, not counseled on breast feeding, home delivery, and cesarean delivery were identified as determinants of pre-lacteal feeding. Thus, emphasis should be given to improve mothers’ Infant and Young Child Feeding practice through counseling and utilization of institutional delivery. Moreover, special attention should be given to mothers from pastoralist regions and poor socio-economic status to reduce pre-lacteal feeding practice.