Objective: In this study, 5-fluorouracil as the drug model molecules, and gelatin as the carrier of matrine. Methods: The gelatin nanoparticles were prepared by coacervation method, the surface morphology and particle size of the gelatin nanoparticles were observed by Transmission electron microscope (TEM) and Dynamic light scattering (DLS), it's envelopment rate and drug loadings were calculated and the sustained-release effect were detected in vitro. Results: The gelatin nanoparticles had a good surface morphology, the average particle size was 65.1?2.1 nm, dispersed uniformly. The envelopment rate and drug loadings of 5-fluorouracil/ gelatin were 23.5?1.9 % and 69.7?0.5 % respectively. In vitro release experiments displayed that the release of 5-fluorouracil/gelatin nanocomposite accorded with the Higuchi equation, indicating that the gelatin could sustain the release of 5-fluorouracil. MTT measurement showed that the 5-fluorouracil/gelatin nanocomposite has significant cytotoxicity on human gastric cancer cells (SGC7901). Conclusion: The prepared 5-fluorouracil/gelatin nanocomposite has good properties of sustainab release and cytotoxicity and could be used as a kind of drug-loading carrier in the pharmaceutical field.