| Polygonatum odoratum (Mill.) Druce, a Chinese herbal medicine knowed as Rhizoma Polygonati Odorati (RPO), was used as test materials. Respectively, the extraction and comparative analysis of polysaccharide from defferent areas were studied; the antioxidant effect in vitro of polysaccharide was investigated, and the contents of total saponins and diosgenin were determinated. Besides, the tissue culture and rapid propagation were researched. The main results were as follows:1. Comparison of the content and the antioxidant activity of polysaccharide. The polysaccharide was extracted with methods of hot-water extraction, and the Sevag method was used to remove protein. The content of polysaccharide was determined with the method of the phenol-sulphuric acid and the contents of polysaccharide in RPO derived from ten producing areas were compared. The results showed that the contents of polysaccharide in RPO from different places were significantly different, and that from Jiangxi was the highest (11.89%), while that from Hebei was the lowest (6.37%). The contents of polysaccharide from different areas are Jiangxi > Sichuan > Hunan > HeilongJiang > Neimenggu > Shaanxi > Anhui > Guangxi > Shanxi > Hebei.In addition, the effect in vitro of scavenging free radicals of polysaccharides from RPO was researched. The scavenging ability of polysaccharides on superoxide anion (O2-?), hydroxyl radical (?OH), 1, 1-diphenyl-2-picrylhydrazyl (DPPH?) and hydrogen peroxide (H2O2) were studied in vitro chemical systems. The results showed that the antioxidant effects on O2-? and H2O2 were significant and relatively weaker scavenging activity were showed on ?OH and DPPH?.2. Studies on total saponins and diosgenin of RPO. The total saponins and diosgenin were extracted, and the content of the total saponins was determined by the method of HPLC. The method of the reversed phase HPLC determination of diosgenin was developed. The optimal HPLC conditions were as follow: C18 column as the chromatographic column, methanol-water (88:12, V/V) as the mobile phase, 1mL/min of flow rate, wave length of 210nm in UV detector. The results showed that the contents of the total saponins in RPO from different areas were obviously different, and that from Sichuan was the highest (3.63%), and that from Heilongjiang was the lowest (1.65%). The comparision of the total content saponins was as following: Sichuan > Hebei > Hunan > Neimenggu > Guangxi > Shanxi > Anhui > Jiangxi > Shaanxi > Heilongjiang. The content of the diosgenin was low, only 0.031%-0.053%.3. Tissue culture and rapid propagation of P. odoratum. The explants from the rhizome, the leaf blade and the stem of P. odoratum were cultured on MS media containing different phytohormons. The results showed that rhizome tissue was the most ideal explant for inducing calli and buds, while it was difficult to induce callus from leaf blade tissue or stem tissue. In the culture of rhizomes, MS + BA 1.0 mg·L-1 + NAA 0.5 mg·L-1 was the optimal medium for callus induction, both MS+BA 2.0 mg·L-1+IBA 0.5 mg·L-1 and MS + BA 3.0 mg·L-1 + NAA 0.1mg·L-1 were suitable for the induction of calli and the differentiation of advantitious buds, and the latter one was more suitable for the differentiation and the growth of bud. 1/2MS + NAA 3.0 - 5.0 mg·L-1 was the ideal medium for root induction. The induction rate of callus, bud and root are 87%, 90% and 99% respectively. The survival rate of plantlets after transplanting was more than 85%.By comparing the content of polysaccharide from different areas, a standard of evaluating quality of P. odoratum may be developed. In our studies, an antoxidant system of polysaccharide and the optimal tissue culuture condition of P. odoratum were established, in vitro. The contents of the total saponins in RPO from different areas were found to be obviously different. A method of diosgenin determination by HPLC was established.
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