OBJECTIVE： To provide reference for eliminating the safety accidents occurred in the production process of Amikacin sulfate injection （called “Amikacin” in short）. METHODS： The safety assessment index system of Amikacin production process with hierarchical structure was established， and the weight of each index was calculated by analytic hierarchy process （AHP）. Then， fuzzy comprehensive evaluation （FCE） was used to assess the safety of Amikacin production process in a large-scale pharmaceutical enterprises from southwest China. The improvement suggestions were put forward according to assessment results （as standard score of related index）. RESULTS & CONCLUSIONS： The safety evaluation index system of Amikacin production process is divided into 3 levels of target layer， criterion layer and index layer； criterion layer included 6 first indicators of personnel factor， technological factor， material factor， equipment factor， environmental factor and management factor； index layer included 24 secondary indicators of staff quality， process complexity，inflammability，equipment location， workshop internal environment and safety management，etc. AHP demonstrated that the weight of 6 first indicators were 0.291， 0.066， 0.109， 0.109， 0.045， 0.308， respectively. FCE showed that standard scores of personnel factor，process factor，equipment factor and management factor were higher than others， being 0.804，0.828，0.816，0.818， respectively， which indicated good safety. The standard scores of material factor and environmental factor were lower， being 0.772 and 0.695， which indicated general safety. On the whole， standard score of FCE assessment of Amikacin production process was 0.828， indicating good safety. Because the safety assessment of material factor and environmental factor are lower than others， it is suggested to strengthen the valves， flanges and gaskets check and real-time monitoring， and improve the lighting and ventilation facility.