The interaction between arbekacin and the hemodialysis membrane is unclear. The aim of this study was to evaluate the adsorption of arbekacin (ABK) onto hemodialysis membranes and to establish a simpler system for evaluating the adsorption properties of the drug. First, small cut pieces of polysulfone or sulfonated polyacrylonitrile (AN69) hollow fiber membranes were stirred in a solution of ABK for the qualitative assessment. Then, we designed the experimental system that was approximately 1/100 of the actual size using a small dialyzer “mini-module” of our original design for the polysulfone or AN69. We circulated ABK solution in this system for the quantitative assessment. Finally, we administered ABK in subjects undergoing hemodialysis as a clinical trial to evaluate the adsorption of ABK onto polysulfone or AN69. In the qualitative assessment, the rates of ABK adsorption onto polysulfone and AN69 were 6.2 ± 2.9 and 49.8 ± 1.8 %, respectively. In the quantitative assessment, although there was almost no change in the circulating ABK concentration with the use of polysulfone, there was about 68 % decrease in the circulating ABK concentration with the use of AN69. In the clinical trial, clearance by the dialyzer using AN69 was high at 10 min after starting dialysis, despite the sieving coefficient being 0. However, reduction of ABK clearance was observed with time; the removal rate of ABK at the completion of the dialysis using polysulfone or AN69 was about 67 %, with no difference between the two membranes. The main limitation of this study was the small sample size in the clinical trial. These findings suggest that ABK is adsorbed onto AN69. Furthermore, the present adsorption experiment with a mini-module was considered useful as an evaluation system because it was easy to handle, using less solvent and drugs compared with a previous report, and it reflected the results of the clinical trial.