Using high-pressure liquid chromatography(HPLC) method to determine the saponin constituents of steamed Panax notoginseng. Methods: By HPLC, with 18 alkyl silane bonded silica as filler, and acetonitrile-water as mobile phase for gradient elution, we established a method that could simultaneously analyze 16 notoginsenoside controls(Rg1, Re, Rb1, Rc, Rb3, Rd, Rg3, Rh1, Rg6, F4, Rh4, Rk3, Rk1, Rg5), and measured the constituents of steamed Panax notoginseng. Results: By comparing each peaks retention time of the steamed Panax notoginseng HPLC map with mixed samples HPLC map, we found several saponin constituents (Rg3, Rh1, Rg6, F4, Rh4, Rk3, Rk1, Rg5) which are not produced by unsteamed Panax notoginseng. Conclusion: By HPLC analysis, we confirmed that steamed Panax notoginseng contain several immune-enhancing constituents, which is in accordance with the traditional Chinese medicines theory about Panax notoginseng. This research lay the foundation of future fingerprint research of Panax notoginseng.