An update of practical aspects of the use of labeled water for the measurement of total body water (TBW) and energy expenditure (EE) is presented as applied in Maastricht, The Netherlands. We use a 10-hour equilibration period. The isotopes for the measurement of TBW and EE are routinely administered, after collecting a background urine sample, as a last consumption before the night. Our data show an underestimate of TBW measured with isotope dilution after 4 hours (in the morning), a discrepancy which increases with the size of TBW. No such relation and no significant differences were found after 10-hour (overnight) equilibration. The ratio between the dilution space for deuterium and oxygen-18 is higher than the earlier figure of 1.03, especially in adult subjects with a high body fat content. For an observation period of EE over two weeks, samples from the second and the last voiding on the first, mid, and last day of the observation period are collected. Differences in EE calculated from morning and evening samples within the first and second week allow detection of sampling errors and if so, samples are excluded from the final calculation. Differences of EE between weeks 1 and 2 allow a check for the consistency of the subjects' physical activity level and usually fall within 10% of the average EE over the total observation interval.