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The Research on Chemical Components and Biological Activity of Shandan Huangshen

Author: XueHongYan
Tutor: YangLin
School: Lanzhou University of Technology
Course: Biochemistry and Molecular Biology
Keywords: Huangshen chemical constituents biological activities extracting techniques
CLC: R285
Type: Master's thesis
Year: 2011
Downloads: 56
Quote: 2
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Abstract


Huangshen”is the dried root and rhizome of Sphallerocarpus gracilis(belongs to the Umbelllferae family), which is used as relieving internal heat and pain, harmoning the circulation of menstrual. It wildly distributed in the northwest and northeast of China. In order to improve the exploiting and using of“Huangshen”, the investigations on the chemical constituents and bioactivities of have been carried out in this paper.The chemical constituents of“Huangshen”have been tested systematically by means of pre-test. It found that it contains amino acid, peptide,and proteins, saponins,saccharides and glycosides,flavoniods,alkaloids,carboxylic,steroids or terpenoids,coumarin,macrolides,volatile oils;there is no phenols,tannin,anthraquinones and cardiac glycosides.And then studied bioactivities of the polysaccharides and total saponin: (1) MTT colorimetric assay was adopted to test their anti-tumor activities to human hepatoma cell lines Hep-G2. The results showed that the polysaccharides and total saponin of Huangshen both exihibited no significant inhibitory effects on tumor cells. (2) Their antimicrobial activities in vitro ware evaluated by filter paper method. The results showed that the polysaccharides had no inhibitory effects to the four bacteria (Staphylococcus aureus, Eschercihia Coli, Streptococcus uberis, Klebsiella pneumoniac), while the total saponin showed different inhibitory effects to the four bacteria, and it showed the best inhibitory effects to Staphylococcus aureus. (3)The scavenging effects on hydroxyl radicals and superoxide anion radicals of the polysaccharides and total saponin were studied with Vc as a positive control. The results showed they both had the scavenging activities to hydroxyl radicals and superoxide anion radicals, and the reducitive potential in a concentration-dependent manner. Among which the scavenging capacities of the polysaccharides on hydroxyl radicals and superoxide anion radicals were lower than the capacities of Vc, the scavenging capacities of total saponin were correspond with Vc.Because of total saponin of Huangshen had significant inhibitory effects to bacteria and the scavenging activities to hydroxyl radicals and superoxide anion radicals, according to the total saponin extracting rate, the extracting techniques of total saponin were optimized by single factors and orthogonal design. The optimum extracting condition was showed as follows: the concentration of ethanol was 80%, extracting temperarure was 70℃, solid-liquid ratio was 1:20, extracting time was 150 min.Seven compounds were isolated from the petroleum ether (60-90℃) and ethyl acetate soluble part of methanol extract by means of column chromatography(CC), thin layer chromatography(TLC), recrystallization, etc, and five out of seven were elucidated as apoil(Ⅱ), stearic acid(Ⅳ),β-sitosterol(Ⅴ),daucosterol(Ⅵ) andβ-D- fructopyrano-(1→4)-O-β-D-glucopyranose(Ⅶ)by modern spectroscopic methods (1H-NMR, 13C-NMR, EI-MS etc.), along with comparison to authentic samples or the data from literatures.Results of the chemical compositions and biological activities in the thesis provided scientific theoretical evidences for using of Huangshen medicines, and exploiting and using of“huangshen”.

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