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Background: Neonatal jaundice is one of the most common neonatal problems especially in preterm ones. Preterm babies are treated with different types of phototherapy. Preterm neonates have a weak antioxidant defense mechanism. Both the hyperbilirubinemia and phototherapy have an impact on this antioxidant defense system. Objective: In the current study we aimed to assess the effect of hyperbilirubinemia and phototherapy in the oxidant/antioxidant status of preterm newborns. Subjects and Methods: Thirty late preterm (≥35 weeks) neonates with non-hemolytic neonatal jaundice in the 3-9 days of life were involved in the preset study, and received phototherapy. 15 healthy neonates of the same gestational and postnatal age without jaundice were included as a control. We assessed the total oxidant status (TOS), total antioxidant capacity (TAC), and oxidative stress index (OSI). Results: TOS and OSI were significantly higher (P<0.01); TAC level was significantly lower (P<0. 01); in jaundiced preterm neonates than in control group. Following phototherapy there were significant increase in TOS and OSI (P<0. 01) and decrease in TAC (P<0.001). Conclusions: Both hyperbilirubinemia and phototherapy induce oxidative stress in preterm neonates. [ABSTRACT FROM AUTHOR]