Aims: The higher ultrafiltration (UF) induces poor outcomes. The impact of higher UF on the volume status was investigated. Methods: 60 hemodialysis (HD) patients were divided into three groups according to the ratio of total UF to post-dialysis body weight (TUF/PDW) (<3%, 3–5%, ≥5%). ANP, the ratio of extracellular water to total body water and excess fluid mass (ExF/PDW) by bioimpedance spectroscopy, inferior vena cava diameter by ultrasound were measured at the end of HD. The ratio of post-HD blood volume to pre-HD (BVpost/BVpre) and standardized filtration coefficients (Lpst) of the microvasculature in the vicinity of PDW were calculated. Results: Only Lpst and BVpost/BVpre showed significant differences among the three groups. A stepwise multiple linear regression model revealed that BVpost/BVpre was correlated with TUF/PDW, ExF/PDW and Lpst (R = 0.778, p < 0.001), independently. Conclusion: Higher UF causes decreases in BVpost/BVpre and Lpst. BVpost/BVpre was determined by TUF/PDW, ExF/PDW and Lpst.