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Bioelectrical impedance analysis (BIA) reference values are based on supine assessments. Little is known regarding the effects of time course shifts in body water compartments after assuming a supine position. The aim of this study was to characterize these effects and provide recommendations regarding the optimal waiting time to perform BIA. Thirty-eight healthy adults underwent BIA via the RJL Quantum Legacy analyzer immediately upon lying down and every 5 minutes for 15 minutes. Differences in resistance (R), reactance (Xc), intracellular (ICW), extracellular (ECW), total body water (TBW), body fat percentage (%BF), and phase angle (PhA) were assessed. There were small but significant increases in R, Xc, and %BF (all p<0.001), as well as small but significant decreases in ICW, ECW, and TBW (all p<0.001) over 15 minutes. No difference was observed for PhA (p=0.065). Average values changed over 15 minutes by +7.14Ω, +1.36Ω, -0.2L, -0.2L, -0.4L, +0.05° and +0.1% for R, Xc, ICW, ECW, TBW, PhA and %BF, respectively. BIA measurements are affected by shifts in body water compartments after assuming a supine position, but these differences lack clinical significance in healthy adults. Technicians working with healthy adults can perform BIA within 15 minutes after participants assume a supine position.