Background: The possibility of vitamin D deficiency is high among mothers and their premature newborns. Objectives: Due to the high rate of vitamin D deficiency in Iranian women, this study aimed to determine the association between the serum levels of vitamin D in the blood of premature newborns and their mothers in the Iranian population. Methods: This cross-sectional study was conducted on 324 mothers and their 324 newborns with gestational age less than 37 weeks, who were referred to Ghaem Hospital affiliated to Mashhad University of Medical Sciences, Mashhad, Iran, during 2016-2020. After clamping, 1.5 cc blood samples were obtained through the umbilical cord to determine the neonates’ vitamin D levels. Finally, the serum levels of vitamin D were measured, and the correlation between the levels of vitamin D of mothers and their newborns was calculated. Results: The mean vitamin D levels of newborns and mothers were 15.75 ± 11.18 and 20.16 ± 13.41 ng/mL, respectively. Moreover, there was a high correlation between the vitamin D levels of mothers and their newborns (r = 0.672; P < 0.001). Furthermore, a significant correlation was observed between the vitamin D levels of mothers, who used vitamin D supplementation during pregnancy and their neonates’ levels (r = 0.773; P < 0.001). Conclusions: The findings suggest a positive correlation between neonatal and maternal vitamin D levels. Moreover, it seems that maternal vitamin D levels can predict neonatal vitamin D deficiency.