The measurement of transepidermal water loss (TEWL) is important for evaluating the integrity of the barrier function of the stratum corneum. Normal TEWL values in healthy adults and in children ages 2 and older are well known, but few studies have been performed in infants and neonates. TEWL in healthy neonates younger than 24 hours old was assessed and compared with that of an adult study population. We also studied possible correlations between this parameter, gestational age, and mode of delivery. A prospective study was conducted in healthy newborns. The areas tested were the volar forearm and the popliteal fossa. Ninety-nine healthy newborns were enrolled and 33 healthy adults were analyzed as controls. Statistically significant differences were noted between newborns and adults in TEWL (p < 0.01). Newborns had a much higher mean TEWL than adults. Differences in the morphology and physiology between newborn and mature skin can explain the higher TEWL in newborns. Higher TEWL could also be due to the sudden functional adaptation of the skin immediately after delivery, when the newborn transits from a liquid to the dry, gaseous extrauterine environment. Functional evaluation of the neonatal skin barrier is important mainly because maintaining skin integrity facilitates cutaneous adaptation.