Objective:To prepare 5-fluorouracil(5-FU) magnetic liposome with Fe/Fe3O4 core-shell structural magnetic nanoparticles(FCSN) and analysis its characteristics. Methods:Fe3O4 nanoparticles were used to prepare FCSN by reduction-oxidation method. The characteristics of FCSN were determined by X-ray diffraction (XRD), transmission electron microscopy (TEM) and vibrating specimen magnetometer (VSM). Using reverse-evaporation method to prepare 5-FU-FCSN magnetic targeted liposome and the best formula of magnetic liposome was obtained through the four factors of three levels orthogonal experiment. The characteristics of magnetic liposome were measured with particle size analyzer(PCS) and TEM. Entrapment efficiency was determined by gel-chromatography. Results:FCSN was similar to a globe with the average particle size of 70 nm. XRD and TEM observation of FCSN shown that its shell was Fe3O4 and kernel was Fe, magnetic saturation intensity was 107.54 emu/g. 5-FU-FCSN magnetic liposome was similar to a globe with the average particle size of 202.5 nm, the entrapment efficiency of 33.5%. 5-FU-FCSN magnetic liposome was relatively stable in 4℃ refrigerator after a month. Conclusion:5-FU-FCSN magnetic liposome was successfully prepared, which can be stored in 4℃ refrigerator for a long time.