Abstract: Objective To explore the levels of serum calcium, phosphorus, alkaline phosphatase and vitamin D of premature infants with different gestational weeks, weights and born in different seasons. Method A total of 488 premature infants were collected as objects between June 2018 and July 2019 from Neonatology department in Yangzhou Maternal and Child Health Hospital.According to the gestational age at birth, premature infants were separated as 27-<32 weeks group, 32-<35 weeks group and 35-<37 weeks group. According to the birth weight, they were separated as very low birth weight group, low birth weight group, nomal birth weight group. According to the birth season, they were separated as winter and spring group, summer and autumn group. The levels of serum calcium, phosphorus, alkaline phosphatase and vitamin D of different groups were compared. Result The levels of serum calcium, phosphorus and alkaline phosphatase of premature infants were associated with the gestational age at birth, birth weight and birth season. However, the levels of vitamin D of premature infants were not associated with the gestational age at birth, birth weight and birth season. Conclusion The levels of serum calcium, phosphorus and alkaline phosphatase of premature infants were associated with the gestational age at birth, birth weight and birth season. It is necessary to monitor the levels of serum calcium, phosphorus and alkaline phosphatase of premature infants to provide early intervention.