Panax notoginseng was used as the medium for lactic acid bacteria fermentation to manufacture product with antihepatocarcinoma activity. The fermentation broth prepared in a 250 mL Erlenmeyer flask was found to possess antiproliferation activity against hepatoma Hep3B cells. At the dosage of 500 microg/mL, the viability of hepatoma Hep3B cells was approximately 2.2%. When the fermentation was scaled up to a 6.6 L fermenter, it was found that the fermentation broth produced at 37 degrees C for 2 days showed the highest antihepatoma activity. Animal study revealed that when Hep3B implanted SCID mice were treated with 1000 mg/kg BW/day of the fermentation broth, tumor volume and tumor weight were reduced approximately 60% as compared to the negative control group. HPLC analyses showed that saponins in P. notoginseng including notoginsenoside R(1) and ginsenosides Rg(1), Rb(1), Rd, and Rh(4) decreased, but ginsenosides Rh(1) and Rg(3) increased during fermentation. LC-MS/MS revealed that the minor saponins ginsenoside F(1), protopanaxatriol, and notoginseng R(2) also exist in the fermentation product. It appears that ginsenoside Rg(3), ginsenoside Rh(1), and protopanaxatriol are possibly responsible for the enhanced antihepatocarcinoma activity of the P. notoginseng fermentation broth.