Abstract: OBJECTIVE To prepare tubeimuside A long-circulating liposomes(TA-LC-Lipo), and to study the in vitro properties of TA-LC-Lipo. METHODS TA-LC-Lipo was prepared by ethanol injection; encapsulation efficiency and in vitro release rate of TA-LC-Lipo were determined by HPLC; particle size and Zeta potential of TA-LC-Lipo were measured by a Marlvern Zetasizer Nano-ZS90; hemolysis of TA-LC-Lipo was assessed by visual observation and UV analysis. RESULTS The optimum prescription was as follows:the concentration of soybean phospholipid(SPC) was 10 mg·mL^-1, the ratio of drug to SPC was 1:10, the ratio of SPC to cholesterol was 4:1, and the molar content of DSPE-PEG 2000 was 5%. The average particle size, PDI, Zeta potential, and encapsulation efficiency of the prepared TA-LC-Lipo were 123.0 nm, 0.134, -1.20 mV, and 86.2%, respectively. CONCLUSION The prepared TA-LC-Lipo shows uniform particle size distribution, high encapsulation efficiency, sustained release, and low hemolytic activity in vitro.