Objective: To investigate the effect of hydroxypropyl-sulfobutyl ether-β-cyclodextrins（HPn-SBEm-β-CD）on hemolysis and morphological changes in red blood cells (RBC) and on nervous system, respiratory system and cardiovascular system in animals. Methods: Hemolysis of 11 different degrees of substitution of HPn-SBEm-β-CD was detected by UV. Scanning Electron Microscopy (SEM) was used to observe the effect of HP3-SBE2-β-CD on morphological changes in RBC. Mouse were given HP4-SBE3-β-CD of 80, 160, and 320 mg/kg of intravenous administration in the experimental groups respectively. Different physical parameters such as common behavior, times of free activities were taken. Beagle dogs were fed anesthetically HP4-SBE3-β-CD of 40, 80, and 160 mg/kg of intravenous administration and were taken the respiratory change of rate, extent and tension, as well as ECG, blood pressure and heart rate. Results: The results of hemolytic activity was to make clear that the haemolyticus of HPn-SBEm-β-CD were all less than β-CD , most of the compounds were less than the HP-β-CD, some of the them were also less than the SBE7-β-CD such as HP2-SBE3-β-CD、HP1-SBE4-β-CD and HP3-SBE6-β-CD. 10 mM HP3-SBE2-β-CD had no effect on morphological changes in RBC, but 2.5 mM M-β-CD had changed it obviously. HP4-SBE3-β-CD of 80, 160, and 320 mg/kg had no effect on behavior and times of free activities. HP4-SBE3-β-CD had no obvious effect on the respiratory change of rate, ECG, heart rate and blood pressure. Conclusion: HPn-SBEm-β-CD with low hemolysis has no effect on the nervous system of mouse, and also has no effect on respiratory system and cardiovascular system of anesthetical dogs.