The three-compartment (3-C) model of physique assessment (fat mass, fat-free mass, water) incorporates total body water (TBW) whereas the two-compartment model (2-C) assumes a TBW of 73.72%. Deuterium dilution (D2O) is the reference method for measuring TBW but is expensive and time consuming. Multifrequency bioelectrical impedance spectroscopy (BIS SFB7) estimates TBW instantaneously and claims high precision. Our aim was to compare SFB7 with D2O for estimating TBW in resistance trained males (BMI >25kg/m2). We included TBWBIS estimates in a 3-C model and contrasted this and the 2-C model against the reference 3-C model using TBWD2O. TBW of 29 males (32.4 ± 8.5 years; 183.4 ± 7.2 cm; 92.5 ± 9.9 kg; 27.5 ± 2.6 kg/m2) was measured using SFB7 and D2O. Body density was measured by BODPOD, with body composition calculated using the Siri equation. TBWBIS values were consistent with TBWD2O (SEE = 2.65L; TE = 2.6L) as were %BF values from the 3-C model (BODPOD + TBWBIS) with the 3-C reference model (SEE = 2.20%; TE = 2.20%). For subjects with TBW more than 1% from the assumed 73.72% (n = 16), %BF from the 2-C model differed significantly from the reference 3-C model (Slope 0.6888; Intercept 5.093). The BIS SFB7 measured TBW accurately compared with D2O. The 2C model with an assumed TBW of 73.72% introduces error in the estimation of body composition. We recommend TBW should be measured, either via the traditional D2O method or when resources are limited, with BIS, so that body composition estimates are enhanced. The BIS can be accurately used in 3C equations to better predict TBW and BF% in resistance trained males compared with a 2C model.