Objective To investigate the preparation of nanoparticles loading the effective components of Compound Huangdai Tablets (Arsenic sulfide, Indirubin and Tanshinone) and the therapeutic effect of HepG₂ and Huh-7 cells. Methods The poly (D,L- lactide-co-glycolide) nanoparticles loading Arsenic sulfide, Indirubin and Tanshinone were prepared by o/w emulsification-solvent evaporation method. The drug encapsulation efficiency, loading efficiency and release profile in vitro were measured by Hydride generation atomic absorption spectrometry and High-performance liquid chromatography. The morphology and size of nanoparticles were investigated by scanning electron microscopy and particle size analyzer, respectively. Cell viabilities were measured by MTT assay and forming colonies. Results The nanoparticles were spherical, with a diameter of 115.09 ± 36.71 nm and a polycentrifugation index of 0.131. The drug encapsulation efficiency and loading efficiency of Arsenic sulfide, Indirubin and Tanshinone were 5.17±1.26%, 1.03±0.45%, 1.72±0.67% and 16.15±4.7%, 76.74±6.8%, 83.09±9.4%, respectively. The mimics the drugs release in vitro were biphasic with early bursting release followed by slow and sustained release. Fluorescence microscopy revealed phagocytosis of nanoparticles by HepG₂ and Huh-7 cells. MTT detection and cell survival assay showed decreased cell viability after treatment with free drugs and drug loading nanoparticles compared with control cells, and drug loading nanoparticles were more significant than free drugs. Conclusions The nanoparticles loading Arsenic sulfide, Indirubin and Tanshinone have definite therapeutic efficacy on hepatocellular carcinoma, providing a new dose type for the clinical administration of Traditional Chinese Medicine.