To determine whether infusion of 13C-labeled bicarbonate can be used to measure rates of CO2 production (VCO2), seven healthy adults received 6-h primed continuous intravenous infusions of NaH13CO3 and L-[1-14C]leucine in the post-absorptive state while VCO2 was measured by indirect calorimetry. Indirect calorimetry and the use of specific activity and rate of 14CO2 expired yielded identical values of VCO2: 8.97 +/- 0.82 and 8.80 +/- 0.83 mmol/min, respectively (P = NS). The concentration of NaH13CO3 in the infusates and the 13C enrichment in breath CO2 were determined using gas chromatography-isotope ratio mass spectrometry. The rate of appearance of CO2 measured using the NaH13CO3 infusion rate and the steady-state breath 13CO2 enrichments was 11.41 +/- 1.56 mmol/min, which was higher (P < 0.001) than that determined by either of the other two methods. When corrected for the recovery of labeled CO2 during the infusion of NaH13CO3 by use of published values, rate of appearance of CO2 was 9.24 +/- 0.78 mmol/min, which did not differ from VCO2 determined using the other two methods. We conclude that infusion of NaH13CO3 can be used to determine VCO2. This method should be useful to study the oxidation of substrates in populations such as ventilator-dependent neonates, in whom indirect calorimetry is laborious and inaccurate.