Background: There is still no consensus on the optimal infusion flow (Qi) in mid-dilution hemodiafiltration. The aim of this study was to compare mid-dilution with varying Qi. Subjects andMethods: Prospective study in 25 patients who underwent seven hemodialysis sessions with a Qi of 0, 50, 100, 150, 200, 250 and 300 ml/min. Results: All sessions were well tolerated except Qi 300 ml/min. No significant differences in urea, creatinine, α1-microglobulin or α1-acid glycoprotein reduction ratios were observed. β2-Microglobulin, myoglobin and prolactin reduction ratios were higher with Qi 150, 200, 250 and 300 ml/min in comparison with Qi of 0, 50 and 100 ml/min. There were no differences in the removal of small or larger molecules when Qi was 150 ml/min or higher. Conclusions: Optimal Qi in mid-dilution appears to be in the range of 150–250 ml/min since good clinical outcomes, similar efficiency and no technical complications up to a Qi of 250 ml/min were observed.