Deuterium or tritium labeled water traditionally has been used for the measurement of total body water by application of the dilution principle. However, these methods have not enjoyed wide clinical use. The use of deuterium is hampered by the tedious and time consuming nature of the analysis while the use of tritium involves a radiation hazard. In addition, exchange of the label with nonaqueous hydrogen in the body raises questions about the accuracy of total body water values. To determine if water labeled with the stable isotope 18O can yield faster and more accurate results, total body water was measured simultaneously using water labeled with both 18O and 2H. The 18O and 3H dilutions were measured by mass spectrometry. The relative precision of the body water value using the 18O method was 2% for both serum and breath analysis. The 18O was fully equilibrated within 2 to 3 hr after administration, and results from the analysis of breath CO2 could be readily obtained within 1 hr after sampling. The H2(18)O dilution space averaged 3.0% (SE = 0.4) less than the 2HHO dilution space, because the latter exchanges with nonaqueous hydrogen. For this reason, the H2(18)O dilution should be a more accurate measure of total body water than the 3HHO dilution.