For the purpose of clinical intravenous injection of 10-hydroxycamptothecin, a novel formulation of lipid nanoparticles loaded with 10-hydroxycamptothecin-phospholipid complex (HCPT-PC-LNs) was prepared by solvent evaporation and high-pressure homogenization methods. Spherical particles with a mean particle size of 200 nm and high encapsulation efficiency of 97.39 +/- 0.91% could be achieved under optimal conditions. In vitro release profile showed that the release pattern of HCPT from nanoparticles was retarded with neglectable initial burst release and well-fitted to Ritger-Peppas equation. In vivo distribution studies in mice showed that HCPT-PC-LNs were mainly localized in liver. Besides, in situ mouse hepatoma model was established and tumor uptake of HCPT-PC-LNs was much higher than that of free HCPT (more than 18-fold). It was found that HCPT-PC-LNs exhibited high growth inhibitory effect on human liver cancer cells by MTT assay. The cellular uptake of HCPT was 24.33 +/- 1.30 x 10(-7) microg/cell in HCPT-PC-LNs treated group at 12 h, which was almost 14-fold higher than that of free HCPT (1.72 +/- 0.57 x 10(-7) microg/cell). This study suggested that the HCPT-PC-LNs could be utilized as a novel formulation for liver tumors therapy, which might be applied in clinic in the near future.