Abstract: Based on the joint analysis of EMG spectrum and amplitude method (JASA), a study on muscle fatigue assessment of spinal surgical instruments based on surface EMG signals was carried out, and a comparative evaluation of the operating comfort before and after the optimization of spinal surgical instruments was completed. A total of 17 subjects were recruited to collect the surface EMG signals of their brachioradialis and biceps. Five surgical instruments before and after optimization were selected for data comparison, and the operating fatigue time proportion of each group of instruments under the same task was calculated based on the RMS and MF eigenvalues. The results showed that when completing the same operation task, the operation fatigue time of the surgical instruments before optimization was significantly higher than that after optimization (P<0.05); there was no significant difference in the fatigue status of brachioradialis and biceps when operating the same instrument (P>0.05). These results provide objective data and reference for the ergonomic design of surgical instruments and fatigue damage protection.