The objective of this study was to investigate the proteome composition in pretermand term‐derived human umbilical cord. Umbilical cord samples were collected from 6 preterm infants with gestational age less than 36 weeks or 4 full terms together with medical information during prenatal period. Several biomarkers are routinely used clinically for predicting preterm labor; however, these factors are either nonspecific or detected too late. Protein profiles were performed on samples from both preterm‐ and termderived human umbilical cord by using Two‐dimensional gel electrophoresis (2‐DE). Approximately 200 different proteins were identified between preterm‐ and term‐delivered umbilical cords. Among them, differentially expressed 34 proteins were identified in 48 protein spots. In the preterm‐derived human umbilical cords, 15 proteins were present at higher levels (2.0‐ to 9.28‐fold increases) and 19 were present at lower levels (2.0‐ to 11.8‐ fold decreases or not detectable) compared to the control term‐derived umbilical cords. Proteomics approaches such as 2‐DE could greatly facilitate the discovery of new and better biomarkers. The high sensitivity and specificity achieved by the combined use of the selected biomarkers show great potential for the early detection of Adverse pregnancy outcomes such as pre‐eclampsia, small for gestational age infants, preterm delivery and placental abruption are associated with higher mortality. Increased amount of HIF‐1 α, GAPDH and HSP27 were observed in preterm‐derived umbilical cords was due to hypoxia‐ dependant and oxidative stress‐independent manner. Moreover, we isolated HUVEC (Human umbilical vascular endothelial cells) from preterm‐ and term‐derived umbilical cords and examined LDH activity. The results of the current study may provide important insights into the molecular mechanisms underlying umbilical cord development and also these data will contribute to a better understanding of the composition of preterm‐ and term ‐ derived human umbilical cord and aid the discovery of novel biomarkers for the prenatal diagnosis of fetal abnormalities