The aim of this study was to investigate the grading of diastolic dysfunction (DD) in relation to hemodialysis in patients with end stage renal disease (ESRD) on hemodialysis (HD) Cardiovascular disease is prevalent in patients with ESRD and accounts for significant morbidity and mortality. Left ventricular hypertrophy (LVH) is common in ESRD but little is known about the impact of HD on currently recommended grading schemes for DD. Comprehensive echocardiographic data was obtained in consecutive patients with ESRD before (n = 247) and immediately after (n = 239) standard HD regimen. Grading of DD was performed according to current recommendations both pre- and post HD. Prior to HD, DD was classified as present in 83 patients (34%), indeterminate in 51 patients (21%) and absent in 113 patients (45%). Patients with DD at baseline compared to those without were older [67.3 years (13.1) vs. 63.2 (14.3), p = 0.037], were more likely to have diabetic- or hypertensive ESRD (43.4% vs. 35.4%, p = ns) and LVMi was significantly higher [119 g/cm2 (27.5) vs. 103 g/cm2 (24.3), p < 0.001]. After HD [mean HD time = 221 min (27.6), mean ultrafiltration volume = 2 L (1.1)], 39 patients (16%) exhibited sustained DD. These patients were older [69.4 years (14.5) vs. 65.0 years (13.9), p = 0.071], were more likely to have diabetic- or hypertensive ESRD (59% vs. 36%, p = 0.010). Myocardial adverse remodeling was more advanced with higher LVMi [127.4 g/m2 (27.5) vs. 106.5 g/m2 (25.3), p < 0.001], lower LVEF [44.7% (11.0) vs. 54.5% (8.7), p < 0.001] and more impaired GLS [- 13.4% (4.3) vs. - 15.8% (4.0), p = 0.006]. Echocardiographic evaluation of diastolic function in patients with ESRD on HD is critically dependent on timing relative to dialysis. The presence of sustained DD after volume unloading by HD identifies a population of patients with an adverse phenotype of blunted vascular response and severe cardiac remodeling.