Abstract: The extraction process of the total saponins in the root of Ophiopogon japonicus by ultrasonic wave was optimized through orthogonal test. Then the saponins were separated by the HZ-801 absorbent resin and the purified samples were identified with chemical and chromatographic method. At last, the immunoregulatory activities of total saponins were studied in vitro. The optimal combination parameters of the extracting technology for 100 g tuber of Ophiopogon japonicus were extracting temperature 60 ℃, ethanol volume fraction 70%, the ratio of ethanol to material 15:1, extracting time 2 h, and extracting twice. The yield of total saponins was (0.508±0.23)%. It was found that three different fractions, named ophiopogonin B(15.4%), A(40.6%), and C(12.6%), were proved to be steroidal saponins by chemical test. Saponin, which was purified preliminarily by ether and n-butyl alcohol and sodium hydroxide, had prominent immunoregulatory activities, i.e., remarkable macrophage-activating capability and nitric oxide and interleukin-1 producing ability.

Key words: Ophiopogon japonicus(L.f.)Ker-Gawl, saponin, separation, identification, immunoregulatory activity