The purpose of this study is to investigate the stationarity of the electromyographic signal in various flexion angles, loads, and window sizes, which influence the result of the mean power frequency (MPF) and median frequency (MNF) analysis. Six healthy subjects participated in the experiment. They were tested in the combination of 3-level flexion angles (0 degree, 22.5 degree, 45 degree) and 3-level loads (0Nm, 30Nm, 60Nm). Electromyographic data were collected for 20 seconds during isometric contraction. The stationarity of collected data were analyzed with four different window sizes including 250, 500, 1000 and 2000ms. Two test methods for stationarity such as Reverse Arrangements Test and Modified Reverse Arrangements Test were used. In order to show the effect of nonstationarity, the increasing/decreasing trend of MPF and MNF trend were discussed. In results, the stationarity of the electromyographic signal decreased as flexion angle increased and load decreased while window size decreased based on Reverse Arrangements Test. The highest stationarity was shown at 500 ms window in Modified Reverse Arrangements Test. The inclination of MNF and MPF indicated 3.6-6.3%, 3.8-5.1% discrepancy compared to the result from stationary data.