Studies On The Geographic Variation Of Chemical Composition And Biological Activities In Rhizoma Polygonati Odorati

Polygonatum odoratum (Mill.) Druce, the perennial herb, was the traditional Chinese medicine. It could be applied in food and drug industry. The plant contains a varity of steroidal saponins, flavonoids, polysaccharides, quinones, lignans and other chemical components, which is significant in Pharmacological. It will be a highly economic value for Polygonatum resources utilization.Predecessors have done more research on Polygonatum, which mainly consentrate in chemical composition, microstructure properties and pharmacological aspects. The studies found that, there were varieties of Polygonatum source which always confused and chemical constituents which have apparently intraspecific variation. The unstable quality and single evaluation methord severely hamper the utilization of Polygonatum and reduce the clinical efficacy.Therefore, in order to ensure the stable quality of Polygonatum and enhance its position in international medicine market, it is necessary to study the chemical components and biological activity variation within Polygonatum species. This study will provide a reference for species choice and quality standard improvement.The main results are as follows:1. Polysaccharide, saponin and flavonoids content of Polygonatum from different origin were mearsured for the first time. Polysaccharide contents in different samples are between 20.3928% and 33.1636%. The highest is Taian sample and the lowest is Anhui sample. Saponin contents in different samples are between 0.0944% and 0.2780%. Saponin of Liaoning sample is highest and Saponin of Heilongjiang sample is lowest. Flavonoids contents in different samples are between 0.0468 % and 0.0931%. Flavonoids contents of Hunan sample are largest and two times as much as Taian sample, which is the lowest. According to polysaccharides, saponins and flavonoids contents, 10 sample were divided into four categories by cluster analysis.Though the sum of three ingredients of groupⅠis largest, which up to 29.27%. But the saponin contents of groupⅠwas smallest in four groups.Ⅲ,Ⅳgroups have lower polysaccharides, flavonoids contents respectively. Considering all the three ingredients Simultaneously, the study show that Polygonatum of groupⅡhave the best quality, the areas of groupⅡare Anhui, Hunan, ShandongQingdao. 2. Polygonatum saponin and flavonoids were separated by macroporous resin, and ingredient contents are better than the solvent extraction method.. The content of flavonoids were 8.24% and the acquisition ratio were 0.174%, which isolated by DM130 macroporous resin. The saponin content and acquisition ratio were 19.11% and 0.81% respectively.3. DPPH assay, the reducing power ability assay, and OH free radical assay were applied to compare the antioxidant active of Polygonatum from different areas for the first time. The result show that flavonoids and saponins exhibited best antioxidant ability to DPPH assay and the reducing power ability, followed with water extract, 70% ethanol extract and 70% alcohol polysaccharides. 70% alcohol polysaccharides has good scavenging ability to OH free radical. For different Polygonatum sample, Liaoning, Hunan, Anhui, ShandongTai'an, ShandongQingdao five flavonoids sample are at top rank in DPPH assay and the reducing power ability assay. IC50 of DPPH assay respectively are 33.96μg/ml,37.44μg/ml,38.02μg/ml,38.15μg/ml,38.65μg/ml. The absorbance of the reducing power ability assay respectively are 0.968,0.806,1.066,1.545,1.272 at concentration 0.1mg/ml. ShandongQingdao, Hunan 70% alcohol polysaccharides have superior OH scavenging activity to other species. IC50 respectively are 0.44mg/ml and 0.45mg/ml. Additionally, Guangxi, Liaoning, Sichuan, Anhui, Shaanxi Polygonatum polysaccharides also exhibits excellent OH scavenging activity with IC50 0.52mg/ml,0.53mg/ml,0.54mg/ml,0.56mg/ml respectively.4. 6 species of bacteria and 13 species of plant pathogen were selected out to test inhibition of Polygonatum extracts. In antibacterial experiment, there are obviously effective when the methanol extracts are imposed on Bacillus aerogenes and Bacterium prodigious, especially the inhibition from Hunan sample which is the best of all, and the suppressive zone is more than 15mm at concentration 10mg/ml. The inhibitory zones measured by Hunan flavonoids against Staphylococcus aureus and Bacterium prodigious arrive at 14mm, which shows the sample has a heavily impact on the two bacteria. The conclusion that there were not obvious effects was drawn according to the bacteria inhibitory experiments of Saponin compounds from different origins. Hunan Saponin compounds could suppress the growth of Bacillus aerogenes and Bacterium prodigious, and its diameter was nearly maintained at 8mm. In plant pathogen inhibition experiment, different Polygonatum methanol extracts could inhibit growth Cucumber anthrax pathogen with inhibition rate 55.6% to 74.0%, respectively. The inhibition rate 72.7% of Henan methanol extracts and the inhibition rate 74.0% of Liaoning, Guangxi, Shaanxi methanol extracts were prominent in tested samples. Poplar Canker Pathogen (Xingping) was strongly inhibited by either Polygonatum flavonoids with rate 76.2% to 88.2%. There were not obvious effects were drawn according to Saponin plant pathogen inhibitory experiments. The rate of Henan Saponin to Exserohilum turcicum and Cucumber anthrax pathogen, the rate of Liaoning Saponin to Camellia anthrax pathogen are both up to 45%.5. TLC and HPLC method were used to investigate difference of different Polygonatum flavonoids, and the HPLC Chromatograms were comtected with antioxidant of flavonoids to investigate the correlation of two. TLC analysis showed that: 5 spots were separated from Henan saponins, and Rf respectively are 0.17, 0.38, 0.61, 0.81 and 0.89. Only 2 spots were separated from Henan saponins with Rf 0.17 and 0.61. Saponin reference compounds 3 and 4 were encountered in all samples, and diosgenin appear in Hunan Polygonatum. There are 8 kinds of flavonoids in Hunan and Guangxi Polygonatum which is the most in all tested samples, and Rf are 0.17, 0.36, 0.39, 0.49, 0.59, 0.63, 0.73 and 0.77. Henan Polygonatum flavonoids have the maximum difference with each other. Flavonoids B appears in samples except Heilongjiang. Similarities of flavonoids HPLC Chromatograms were between 0.6 and 0.9, and significant difference wae showed. 2, 3, 4, 5, 6, 7, 8, 10, 11, 13 and 14 peaks were positively correlated with DPPH scavenging effect, and 3, 4, 5, 8 and 11 peaks play a major role. 2, 4, 7, 9, 10, 11, 12 peaks correlated with the reducing power ability. It can be seen that, 2, 4, 7, 11 peaks are important in both two trials.Through studies of chemical composition and biological activity of different Polygonatum, it clearify that how geographical factors affect Polygonatum quality changes. Preliminary study was done on the relationship between biological activity and chemical compositions. It will provide a reference for the distribution centers investigation and quality standards improvement.