Rationale: Current estimation of body fluid volumes in hemodialysis patients using bioimpedance analysis assumes constant specific electrical characteristics of biological tissues despite a large variation in plasma Na⁺ concentrations [Na⁺], ranging from 130 to 150 mmol/L. Here, we examined the potential effect of variable [Na⁺] on bioimpedance-derived volume overload. Method: Volumes were calculated from published whole-body extra- and intracellular resistance data and relationships using either “standard” or “revised” specific electrical characteristics modeled as functions of [Na⁺]. Result: With “standard” assumptions, volumes increased with increasing [Na⁺]. The increase in volume overload was about 0.5 dm³ and 3% of extracellular volume per 10 mmol/dm³ of [Na⁺] in a 75 kg patient. This increase was abolished when the same bioimpedance data were analyzed under “revised” conditions. Discussion: The overestimation in extracellular volume overload in the range of 0.5 dm³ per 10 mmol/dm³ [Na⁺] perfectly matches the positive relationship determined in a large cohort of hemodialysis patients. The bias may be considered moderate when interpreting data of individual patients, but may become important when comparing data of larger patient groups. The bias disappears when analysis of bioimpedance data accounts for differences in tissue electrical properties, using individual [Na⁺].