The invention discloses a test method of the aperture distribution of a hemodialysis membrane. The method comprises the following steps: dissolving one or more solutes in ultrapure water to form a simulation solution, and filtering the simulation solution through the hemodialysis membrane; testing the concentration of corresponding solute molecular weight in the simulation solution and the concentration of corresponding solute molecular weight in a filtrate; calculating a curve of the molecular diameter of the corresponding solute molecular weight and the interception rate of the hemodialysis membrane; establishing the aperture distribution function and the accumulated distribution function of the hemodialysis membrane; and carrying out nonlinear fitting on the accumulated distribution function on practically measured interception rate curve data through least squares fitting analysis software, determining the aperture distribution parameter, and substituting the aperture distribution parameter into the aperture distribution function to obtain the aperture distribution of the hemodialysis membrane. The method is used to rapidly detect the aperture distribution of the hemodialysis membrane., and has the advantages of high accuracy of measured data, simplicity and high practicality.