Objective: Our study aimed to determine interaction of residual urine volume with dialyzer membrane flux on uremic toxicity in HD patients. Methods: This is a retrospective case-control study. Two hundred and five patients on maintenance hemodialysis in our blood purification center were included and divided into two groups of 24 h-urine volume<300 ml and 24 h-urine volume ≥ 300 ml. The patients were further divided into high-flux and low-flux dialysis groups according to the type of dialysis membrane used in each group. Baseline demographic, clinical data and laboratory values were recorded and compared in each group. Results: Patients with 24 h-residual urine volume ≥ 300 ml preferred a higher dialysis frequency (P>0.031). Body mass index(P=0.015) were significantly higher in patients with 24 h-residual urine volume ≥ 300 ml, in whom serum creatinine(P=0.033), cystatin C(P=0.002), β2-microglobulin(P=0.000) and ferritin(P=0.012) levels were significantly lower. The pre-dialysis β2-microglobulin(P=0.000) and ferritin(P=0.043) levels were significantly lower in the high-flux subgroup in patients with 24 h-residual urine volume < 300 ml, but pre-dialysis intact parathyroid hormone (P=0.049) level and body mass index (P=0.023) were significantly higher in the high-flux subgroup. Levels of serum β2-microglobulin (P=0.439) and intact parathyroid hormone (P=0.190) were not significantly different between high- and low-flux subgroups in patients with 24 h-residual urine volume ≥ 300 ml. Conclusion: High-flux dialysis was more efficient in removal of β2-microglobulin and maintenance of body mass index. Benefits of hemodialysis using high-flux membrane differed in patients with 24 h-residual urine volume < 300 ml and ≥ 300 ml. High-flux dialysis could provide more benefits in HD patients with 24 h-residual urine volume < 300 ml.