Background With the increased survival of preterm babies, retinopathy of prematurity (ROP) has become the leading cause of preventable childhood blindness. It is caused by oxidative damage to the immature retina and zinc may help in the prevention of ROP owing to its antioxidant properties. Objective To measure serum zinc levels in preterm neonates and study its association with the development of ROP. Methods A total of 360 preterm neonates of gestational age between 28 completed weeks to <37 weeks were included in this observational cross-sectional study. Serum zinc levels of the neonates aging <24 hours of life were measured at the time of admission in NICU by spectrophotometry. Taking the reference range of serum zinc levels between 70 to 150 mcg/dL babies were arbitrarily divided into 2 groups (<70 and >70 mcg/dL). ROP screening was done at appropriate postnatal age by indirect ophthalmoscopy. The results were analyzed by the SPSS software version. Results The mean serum zinc level was 114.99±54.47 mcg/dL. The overall prevalence of ROP was 26%; of which, 10% of neonates had severe ROP (≥stage 3). A significant association was found between gestational age and birth weight with ROP. Oxygen supplementation, respiratory distress syndrome, surfactant use, antibiotic use >7 days, and low serum zinc levels (<70 mcg/dL) were significant risk factors on univariate analysis and after multivariate analysis, gestational age, surfactant use, and low serum zinc levels (<70 mcg/dL) came out to be independent risk factors for ROP. Conclusion There was a statistically significant association of low serum zinc levels with the development of ROP and it was independent of other risk factors associated with ROP.