Purpose: We evaluated biochemical analysis results with the aim of discovering serum levels that have possible effects on the development of retinopathy of prematurity (ROP). Methods: A retrospective study was conducted between January 2017 and January 2018 on a total of 110 infants with 35 or less gestational weeks. The sample included 78 infants who had been diagnosed with different stages of ROP and 32 infants without ROP. Results from routine serum biochemical analyses, performed at birth and at one month after birth, were evaluated. The Independent Sample t-test and Mann-Whitney U test were performed to compare the data. Results: The infants with ROP were born at a mean of 28.0 (±2.1) weeks of gestation, weighing a mean of 1066 (±314) g, and the mean duration of stay in the incubator was 38.2 (±19) days. The infants without ROP were born at a mean of 29.6 (±2.6) weeks of gestation, weighing a mean of 1265 (±372) g. 59 infants (53.6%) were females, and 51 (46.4%) were males. Between infants with and without ROP, differences in the following levels were not statistically significant: glucose, blood urine nitrogen, creatinine, alanine aminotransferase, aspartate aminotransferase, albumin, total protein, direct bilirubin, indirect bilirubin, uric acid, and phosphorus. Serum magnesium (Mg) levels at birth in infants with ROP were significantly higher than infants without ROP (p = 0.014). Conclusions: Serum Mg levels at birth were found to be higher in infants with ROP than in those without ROP. The levels may be dependent on the mothers' treatment of Mg for different medical reasons.