The pharmacokinetics of nafcillin were studied in 13 premature neonates with suspected sepsis. The mean weight of the infants studied was 1.19 kg (range, 0.73 to 2.21 kg). Infants less than 7 days of age were given 100 mg of nafcillin per kg per 24 h (every 12 h), and infants more than 7 days of age were given 100 mg of nafcillin per kg per 24 h (every 8 h). Blood samples were obtained before the first dose on day 3 of therapy and at 0.5, 1.5, 3, and 6 h thereafter. Nafcillin concentrations were measured by a microbiological assay. A mean volume of distribution of 326 ml/kg and an elimination rate constant of 0.2040 h-1 were obtained in 10 patients less than 21 days of age. Three patients from 24 to 68 days of age had a mean volume of distribution of 303 ml/min and a mean elimination rate constant of 0.3944 h-1 (P less than 0.05). These data suggest that doses of nafcillin lower than those currently recommended may be adequate to achieve desired peak plasma levels of approximately 75 microgram/ml in infants with low birth weights.