Abstract: OBJECTIVE To explore and analyze the reducing hemolytic effects of puerarin-loaded PEG-PE nano-micelle via the glucose-6-phosphate dehydrogenase (G6PD)-deficient rats blood. METHODS The rat model with G6PD-deficiency was established via injecting 1% acetylphenyl-hydrazine. Then the G6PD-deficient erythrocyte suspension obtained from this rat model was used to evaluate the hemolytic effects of PUE and puerarin-loaded PEG-PE nano-micelle via hemolytic activity, erythrocyte osmotic fragility assay and membrane fluidity of erythrocytes. RESULTS PUE could cause a serious hemolysis to the erythrocyte suspension with the increase of drug concentration and the prolongation of drug incubation time, the hemolytic rate of PUE was up to 38%, while PEG-PEN revealed no significant hemolysis. Additionally, the result of erythrocyte osmotic fragility indicated that puerarin-loaded PEG-PE nano-micelle exerted a slight effect on the erythrocyte membranes, its H_50% was (42.1±1.1)%, much less than that (53.1±1.2)% of PUE. Erythrocyte membrane fluidity indicated that the effect of puerarin-loaded PEG-PE nano-micelle on erythrocyte membrane fluidity was very slight. CONCLUSION The good blood compatibility of puerarin-loaded PEG-PE nano-micelle makes it a favorable candidate for in vivo tests and puerarin-loaded PEG-PE nano-micelle can also provide the useful insight for the further formulation development as an innovative drug.