Objective: To assess application and effect evaluation of 'Five Routine Working Regulation' method in nursing quality management of outpatient injection room. Methods: We invested 16 nurses and 200 patients to join our study in treatment process. Additionally, we grouped the data accord to different time periods. The time periods of control group are from July 2017 to June 2018, we did not implement 'Five Routine Working Regulation' method in this time period. The time periods of intervention group are from July 2018 to June 2019, we implement 'Five Routine Working Regulation' method in this time period. Our researchers collected the information include the quality indicators, nurse satisfaction and patient satisfaction. Result: In the quality indicators research, intervention group participants have better performance than that of control group in this study. In particular, the intervention group has not cases in Total number of no-write opening time of sterile articles in use and Total number of expired or waste of medicine and substance. But the control group has cases in those two domains (0 vs 25, 0 vs 210). In nurse satisfaction research, the result indicate intervention group has better nurse satisfaction assessment in our research result (87.5% vs 62.5%). In patient satisfaction, the intervention group patients provide more good assessment in satisfaction research, and the result is statistical significance (190 vs 171, p = 0.012). Conclusion: the 'Five Routine Working Regulation' method has influence to improve the nursing quality in outpatient injection room.