Performance of the dialysis membrane is strongly dependent upon the physicochemical structure of the membrane. The objective of this study is to devise a new in vitro evaluation technique to quantify the physicochemical structures of the membrane. Three commercial dialyzers with cellulose triacetate (CTA), asymmetric CTA (termed ATA®), and polyether sulfone (PES) membranes (Nipro Co., Osaka, Japan) were employed for investigation. Forward and backward ultrafiltration experiments were performed separately with aqueous vitamin B12 (MW 1355), α-chymotrypsin (MW 25,000), albumin (MW 66,000) and dextran solutions, introducing the test solution inside or outside the hollow fiber (HF), respectively. Sieving coefficients (s.c.) for these solutes were measured under the test solution flow rate of 200 mL/min and the ultrafiltration rate of 10 mL/min at 310 K, according to the guidelines provided by Japanese academic societies. We defined the ratio of s.c. in the backward ultrafiltration to that in the forward ultrafiltration and termed it the index for asymmetricity (IA). The IA values were unity for vitamin B12 and α-chymotrypsin in all three of the dialyzers. The IA values for albumin, however, were 1.0 in CTA, 1.9 in ATA®, and 3.9 in PES membranes, respectively, which corresponded well with the fact that CTA is homogeneous, whereas ATA® and PES are asymmetrical in structure. Moreover, the asymmetricity of ATA® and PES may be different by twofold. This fact was verified in continuous basis by employing dextran solution before and after being fouled with albumin. These findings may contribute to the development of a novel membrane for improved success of dialysis therapy.