Aim: To investigate the relationship between prolactin levels and both gestational age and birth weight in newborn infants. Materials and Methods: 100 eligible antenatal patients were considered for this hospital based prospective study. 3 ml of fetal umbilical cord blood was collected soon after delivery of the baby and cord serum Prolactin levels assayed by Chemiluminescent Micro particle Immunoassay Method (CMIA). Details with antenatal history, intrapartum and postpartum details were collected and entered in an excel spread sheet. Results: The mean serum prolactin levels were highest in neonates born before 28 weeks (691.12 ng/mL) and lowest in those born between 28-32 weeks (511.28 ng/mL). However, the p-value of 0.55 indicates that these differences were not statistically significant. This suggests that gestational age does not have a strong impact on serum prolactin levels in neonates.Neonates with the lowest birth weight (1.5-2 kg) had a mean serum prolactin level of 577.25 ng/mL, while those with a birth weight between 2.5-3.5 kg had a mean level of 526.66 ng/mL. The p-value of 0.32 indicates no significant differences between the groups. This implies that birth weight is not a significant determinant of serum prolactin levels in neonates.The Pearson correlation analysis showed weak correlations between serum prolactin levels and both gestational age and birth weight. The correlation coefficient (r) between serum prolactin and gestational age was 0.08 with a p-value of 0.21, and between serum prolactin and birth weight was 0.05 with a p-value of 0.51. Both correlations were not statistically significant, indicating that there is no strong linear relationship between serum prolactin levels and either gestational age or birth weight in the study population. Conclusion: Overall, the results indicate that while gestational age and birth weight are crucial parameters for neonatal health, they do not significantly correlate with serum prolactin levels in newborns. The majority of neonates were full-term and had normal birth weights, though a significant proportion were preterm or had low birth weights. Despite the physiological importance of prolactin, its levels did not show significant variation with gestational age or birth weight, suggesting that other factors may play a more critical role in determining its levels in newborns.