Abstract Background and Aims Measures of non-invasive pulse wave (PW) analysis can help to assess the status of the central cardiovascular system and to predict clinical outcomes, including morbidity and mortality. During hemodialysis (HD), fluid is extracted from blood. The resulting reduction in relative blood volume (RBV) and collected ultrafiltration volume (UFV) were hypothesized to substantially influence PW characteristics during HD. Volume overload (VO) is common in HD patients and may affect the chance of improving the cardiovascular status of patients during HD. The present study aimed to explore associations between RBV, UFV, VO and a wide range of PW parameters. Method In a single center in Vienna, Austria, 24 patients on maintenance HD underwent four HD sessions each, with whole-body bioimpedance spectroscopy (BIS) before, and automated, cuff-based PW measurements every 15 minutes, RBV- and UFV-monitoring throughout each session. Associations between RBV or UFV as independent variable and 26 different PW characteristics as dependent variables (52 separate models) were tested using confounder-adjusted generalized estimating equations with sandwich estimator and multilevel nested working covariance structure, reporting standardized effect estimates and drop-in-R² to assess variable importance. Sensitivity models including both RBV and UFV as predictors (26 models) were used to assess effects adjusted for confounding and mediation, respectively. The intra-patient correlation between BIS-derived VO and PW parameters at the start of sessions and PW parameter change by end of sessions was assessed using repeated measures correlation analysis. Results Data from 990 PW measurements were available for analysis from an 88% male cohort with median age of 66 years. In half of the PW parameters analyzed, each percent RBV was significantly associated with in summary about 4.3 to 5.7% (95% confidence intervals [CI] 0.9 to 10.46) of the PW parameter's respective standard deviation (SD), negatively so only in heart rate and R-Peak. UFV was negatively associated with a third of PW parameters (effects ranging from −2.5 to −5.47% SD [95% CIs −0.9 to −10.0]), mostly in classical peripheral and central pressures and peripheral resistance. In models including both predictors, RBV overall exhibited greater effect size and variance capture than UFV, except for peripheral resistance. On within-patient level, VO was positively correlated with initial medians of pressure excess (r=0.34, p=0.034), augmentation-index (0.34, p=0.031) and augmentation-pressure (0.32, p=0.042), increase in heart rate (0.32, p=0.04) and decreases in peripheral resistance (0.31, p=0.05), peripheral (systolic: 0.34, p=0.032; diastolic: 0.45, p=0.004; mean: 0.41, p=0.008) and central (diastolic: 0.46, p=0.003) blood pressures during HD. Conclusion The observed PW characteristics were associated with RBV and UFV during HD and may represent modifiable targets for intervention studies. Pressure excess, S-Peak, R-Peak, cardiac and preload parameters were more closely associated with RBV, while classic (especially diastolic) blood pressure parameters, ejection duration and S/D-Ratio were more associated with UFV, in models using both predictors. On average, blood pressure and peripheral resistance tended to decrease less when a patient began HD in a volume-expanded state, which may affect day-to-day feasibility of more classical pressure targets, but perhaps not of other PW parameters. Multiple PW characteristics have previously been shown to be associated with mortality and morbidity and are potential target parameters for future intervention trials. Utilizing RBV modification as an intervention may offer superior precision compared to UFV goals, requiring further studies. PW technology could also be adopted by HD-machine manufacturers at low cost, enabling more precise and individualized treatment options.