Objective: The aim of our study was to determine whether lipid solutions delivered separately or in mixture with total parenteral nutrition (TPN) solutions effect the balance between oxidant and antioxidant levels in premature infants. Methods: A total of 60 preterm newborns who were delivered at their 30–34 gestational weeks and received TPN were included in the study. Premature newborns were randomized into two groups based on the delivery method of the lipid solution, separately (Group 1) or in mixture with TPN solutions (Group 2). Total antioxidant status (TAS), total oxidant status (TOS) and paraoxonase (PON-1) levels were analyzed in both blood samples, and oxidative stress index (OSI) was also calculated. Results: Thirty cases from both groups were included in the study. Statistically significant decrease in the level of TAS and increase in the level of PON-1 were detected at 72 h of TPN therapy in both groups (p < 0.05). Statistically significant decrease in both TOS and OSI levels were observed in Group 2 (p < 0.05). In association with these findings, any statistically significant intergroup difference was not detected in both parameters regarding oxidant balance (p > 0.05). Conclusion: Our study showed that according to lipid administration method any difference for oxidant–antioxidant balance was not detected.