Objective: To determine the effect of a continuous insulin infusion on protein and glucose metabolism in extremely low birth weight (ELBW) infants. Study design: We measured the rate of appearance (Ra) of the essential amino acids leucine and phenylalanine (reflecting proteolysis), utilization of phenylalanine for protein synthesis, and glucose Ra using stable isotope tracers during a basal infusion of glucose (6 mg/kg/min) and in response to a continuous infusion of insulin (0.05 U/kg/hr) by means of the euglycemic hyperinsulinemic clamp technique. Four clinically stable, euglycemic ELBW infants (26 ± 0 weeks' gestation, 894 ± 44 gm birth weight, 2.8 ± 0.8 days of age) were studied. Results: In response to a greater than tenfold increase in insulin concentration (from 7 ± 2 to 79 ± 13 μU/ml), there was a 20% decrease in leucine Ra (Basal: 272 ± 27 μmol/kg/hr; Insulin: 226 ± 29 μmol/kg/hr; p < 0.01) and in phenylalanine Ra (Basal: 91 ± 5 μmol/kg/hr; Insulin: 72 ± 2 μmol/kg/hr; p < 0.05). Use of phenylalanine for protein synthesis also decreased by a similar magnitude (Basal: 77 ± 4 μmol/kg/hr; Insulin: 62 ± 1 μmol/kg/hr; p < 0.05). Glucose utilization doubled (from 8 ± 0.9 to 15.7 ± 1.1 mg/kg/min; p = 0.0003) and plasma lactate concentrations tripled (from 2.1 ± 0.5 to 5.7 ± 1.0 mmol/L; p < 0.05) during the insulin infusion. Conclusions: During an infusion of glucose alone, pharmacologic concentrations of insulin in ELBW infants produced no net protein anabolic effect. Furthermore, euglycemic hyperinsulinemia was accompanied by development of significant metabolic acidosis. (J Pediatr 1998;132:948-53)