Objective To investigate the inhibitory effect and possible mechanism of apoptosis of Yiyi Fuzi Baijiang Powder(YFBS) on hepatocellular carcinoma. Methods C57BL/6 mice model were used to observe the inhibitory effect of YFBS on liver cancer. 24 tumor-bearing mice were divided into model control group, platinum group (5 mg/kg), and low and high dose groups of Yiyi Fuzi Baijiang Powder(7.735, 15.47 g/kg, respectively), treated for 14 d. The effects of different concentrations (10, 20, 30, 40, 50 mg/mL) of YFBS and different treatment time (24, 48, 72 h) on Hepa1-6 cell proliferation were detected by MTT method. Flow cytometry (FCM) was used to detect the apoptosis of hepatoma cells. The levels of Caspase-3, cleaved Caspase-3, Bax and Bcl-2 were detected by western blot. Results The results showed that compared with the model control group, the positive drug group and YFBS groups could inhibit tumor growth of tumor-bearing mice in vivo, with statistically significant differences(P<0.05). Also compared with the blank control group, both the positive group and YFBS groups could significantly inhibit the proliferation of Hepa1-6 cells in a time-dose dependent manner in vitro (P<0.05). The results of flow cytometry showed that YFBS could induce the apoptosis of liver cancer cells, and it had dose-effect relationship. The expressions of pro-apoptotic protein Bax, Caspase-3 and cleaved Caspase-3 increased, while anti-apoptotic protein Bcl-2 decreased, and the ratio of Bax/Bcl-2 increased(P<0.05). Conclusion YFBS had anti-hepatocellular carcinoma effect, inducing carcinoma cell apoptosis by regulating Caspase-3, cleaved Caspase-3, Bax and Bcl-2 protein expression.