We measured the concentrations of cardiac troponin T (cTnT) and amino-terminal pro-natriuretic peptide (NT-proBNP) in umbilical cord blood during the second trimester (20-25 weeks of gestation) and at delivery in 109 uncomplicated pregnancies to define reference values. Using the 97.5th percentile, the upper reference limits for cTnT and NT-proBNP were 0.308 ng/mL and 5402 pg/mL in fetuses in the second trimester, and 0.038 ng/mL and 1690 pg/mL in healthy neonates, respectively. We also evaluated whether labor and delivery mode affected the concentrations of the two analytes. We found significantly higher (p<0.001) umbilical blood NT-proBNP and cTnT levels at 20-25 weeks of gestation than at term. In addition, within the term group, NT-proBNP concentrations were significantly higher in women delivering by elective cesarean section than in women delivering spontaneously (p<0.001), and higher than in women delivering by cesarean section during active labor. This indicates that the decrease in NT-proBNP levels is probably due to labor rather than delivery mode. Finally, we confirmed the at-birth reference values previously established for cTnT and NT-proBNP.