Background: Bioelectrical impedance spectroscopy (BIS) may provide a noninvasive, rapid method for the assessment of total body water (TBW), extracellular water (ECW), and intracellular water (ICW). Few studies, however, have examined the accuracy of BIS in pediatric populations. Objective: Our objective was to evaluate the accuracy of BIS for the measurement of TEW, ECW, and ICW in healthy children. Design: Dual-energy X-ray absorptiometry (DXA), total body potassium (TBK), and BIS measurements were performed in 347 children (202 males and 145 females aged 4-18 y). The reference values for TEW, ECW, and ICW were defined by using a DXA+TBK model. BIS values were evaluated by using the method of Bland and Altman. A randomly selected calibration group (n = 231) was used to derive new BIS constants that were tested in the remaining group (n = 116). Results: BIS values were highly correlated with the reference values (r(2) = 0.94-0.97, P < 0.0001), but differences between the BIS and DXA+TBK models for individuals were significant (P < 0.001). Use of new BIS constants reduced the mean differences between the BIS and DXA+TBK models; the SDs of the mean differences were improved (1.8 L for TEW, 1.4 L for ICW, and 1.0 L for ECW) for the total population. Conclusions: On a population basis, BIS can be calibrated to replace the DXA+TBK model for the assessment of TEW, ECW, and ICW in healthy children. The accuracy of the BIS measurement in individual children may be refined further by using age-and sex-specific adjustments for the BIS calibration constants.