Background: Vancomycin (VCM) is eliminated mainly by diafiltration under continuous hemodiafiltration (CHDF), but the contribution of adsorption to CHDF clearance (CL CHDF ) of VCM using a polyacrylonitrile and sodium methallyl sulfonate copolymer membrane coated with polyethylenimine (AN69ST) or a polymethylmethacrylate (PMMA) membrane is unknown. This study sought to investigate the contribution of diafiltration and adsorption to the CL CHDF of VCM using AN69ST and PMMA membranes in vitro.Methods: An in vitro CHDF circuit model was developed. The initial concentration of VCM was 50 μg/mL and human serum albumin (HSA) was prepared at a concentration of 0, 2.5, or 5.0 g/dL. The effluent flow rate (Qe) was set at 800, 1500, or 3000 mL/h. The CL CHDF , diafiltration rate, and adsorption rate of VCM were calculated.Results: Total CL CHDF of VCM using the AN69ST membrane increased and decreased with increasing Qe and HSA concentration, respectively. Diafiltration and adsorption rates were 82.1 ± 9.8% and 12.1 ± 6.1% under all conditions, respectively. Total CL CHDF using the PMMA membrane increased with increasing Qe. Diafiltration and adsorption rates were 89.2 ± 20.4% and 4.6 ± 17.0% under all conditions, respectively. The observed CL CHDF values significantly correlated with the predicted CL CHDF , calculated according to a previous study as the product of Qe and the plasma unbound fraction.Conclusions: Diafiltration predominantly contributed to CL CHDF of VCM using AN69ST and PMMA membranes. When diafiltration rather than adsorption mainly contributes to the CL CHDF of VCM, the CL CHDF could be predicted from the Qe and HSA concentration, at least in vitro.