To establish normal values for gastric secretory function in preterm infants, we studied 34 healthy preterm infants once a week during hospitalization. Basal acid output, pentagastrin-stimulated acid output, fasting serum gastrin, and fasting serum pancreatic polypeptide were measured during each study. Basal acid output at 1 week of age was 12 mumol/kg/hr, increasing over the first 4 weeks to 30 mumol/kg/hr. Administration of pentagastrin 6 micrograms/kg subcutaneously increased acid output in all age groups. Pentagastrin-stimulated acid output at 1 week was 21 mumol/kg/hr, increasing over the first 4 weeks to 44 mumol/kg/hr. Acid secretion did not change significantly over the next 4 to 6 weeks. Fasting serum gastrin concentration was stable over the first 6 weeks of life, but doubled during the end of the second month. Pancreatic polypeptide was found at low levels throughout the study. These studies confirm that the majority of healthy preterm infants secrete acid in quantity sufficient to maintain the gastric pH less than or equal to 4, providing a barrier to bacteria and protein antigens.