OBJECTIVE： To conduct the abnormal toxicity test and study on the in vitro hemolytic reaction of Gluconate enoxacin injection， and provide reference for its safe use. METHODS： According to “Test for Abnormal Toxicity” in Chinese Pharmacopoeia （2015 edition， Vol.Ⅱ）， half lethal dose （LD50） of Gluconate enoxacin injection and setting of abnormal toxicity limit were carried out in mice. According to “Test for Haemolysis and Agglomeration” in Chinese Pharmacopoeia （2015 edition， Vol.Ⅱ）， study for the in vitro hemolytic and agglomeration reaction was carried out in rabbits. RESULTS： The LD50 of Gluconate enoxacin injection for mice was 248.9 mg/kg； the setting limit of abnormal toxicity test was 28.4 mg/kg， and the results derived from the test using this limit met requirements. The haemolysis and agglomeration results derived from the test using clinical maximum concentration （0.2 g/100 mL） met requirements. CONCLUSIONS： In order to reduce the clinical ADR and feasibility of ensuring quality standard， the abnormal toxicity limit of Gluconate enoxacin injection should be set as 28.4 mg/kg.