Objective To conduct an in vitro wear test of hip joint prosthesis according to ISO 14242, we study the difficult problems such as the fixation and positioning of hip joint prosthesis on the equipment, the determination of lubricant concentration, and the selection of bacteriostatic agent to inhibit the growth of lubricant bacteria. Method Using AMTI hip joint wear tester, different specifications and materials of hip joint prosthesis and different bovine serum bacteriostatic agents were used to conduct multiple wear tests. Results and conclusions The external surface of the acetabular cup needs to be smoothed; For the positioning of the prosthesis, a positioning fixture suitable for the AMTI hip joint wear tester according to the requirements of ISO 14242 was designed; For the determination of lubricant concentration, ultraviolet absorption method or Bradford method is recommended; For the bacteriostatic agent, ProClin 300 can be used as a substitute to test the wear of the prosthesis. Innovation of the article In vitro wear test has a long cycle, complex process, many difficult points in the operation process, and strict requirements for the experimental operation. However, existing research is mostly focused on product application, and there is no detailed research on the test operation method, this study can provide reference for the operators of in vitro wear test of hip joint prosthesis.