Objective? To investigate the effect of umbilical cord ligation time on the incidence of anemia in premature infants, and to provide reference for improving the quality of life of premature infants. Methods A total of 80 premature infants who were treated by vaginal delivery in Huizhou Huiyang Maternity and Child Health Care Hospital from March 2019 to March 2021 were divided into the control group and the observation group by random number table method, with 40 in each group. The premature infants in the control group were treated with traditional umbilical cord ligation, and the premature infants in the observation group were treated with delayed ligation. Both groups were followed up for 6 months after umbilical cord ligation. The blood routine indexes [hemoglobin (Hb), red blood cell count (RBC), hematocrit (HCT)] and transdermal bilirubin at 24 h, 72 h, 1 week and 2 weeks after birth, related iron parameter levels [serum free iron (Fe), total ironbinding capacity (TIBC), serum ferritin (SF)] at 1, 2, and 4 weeks after birth, the occurrence of anemia events at 1, 3, and 6 months after birth were compared between the two groups respectively. Results From 24 h to 2 weeks after birth, Hb, RBC and HCT of premature infants in two groups showed a decreasing trend, but the observation group were significantly higher than those of the control group; from 1 week to 4 weeks after birth, the Fe, TIBC and SF levels of premature infants in two groups showed a downward trend, but the Fe and SF levels of the observation group were significantly higher than those of the control group, and the TIBC level was significantly lower than that of the control group; however, from 1 to 6 months after birth, the incidence of anemia in both groups showed an upward trend, but the observation group was significantly lower than the control group (all P<0.05). From 24 h to 2 weeks after birth, the transdermal bilirubin levels of preterm infants in two groups increased and then decreased, but there was no significant difference between the two groups (all P>0.05). Conclusion Delayed umbilical cord ligation during delivery can significantly improve the blood routine and related iron parameters of premature infants, and reduce the incidence of anemia.