| Objective:To make a preliminary analysis of qualitative and quantitative to Xinjiang Berberis Heteropoda Root chemical composition.On the basis of this,the extract of 5%sulfuric acid from root bark was isolated,purified and analyzed.Finally researched the in vitro antibacterial activity of the chemical constituents of roots.Methods:1.?1?Preliminary qualitative study on the chemical composition of different solvent extracts of root bark and xylem of blackberry was carried out by using the system pretest method of unknown chemical composition.?2?Compared the difference determination of total alkaloids content between ultraviolet spectrophotometry and acid dye colorimetry.And initially determined the total alkaloids content of the root bark and xylem.?3?Investigated and optimized the extraction process of total alkaloids from root bark by Response Rurface Method.2.Using 5%sulfuric acid soaked root bark,then,filtrate gone through cation exchange resin,silica gel column chromatography to separat and purify,after that,combined with NMR,ESI-MS to analyze the structure of monomer compounds,and used HPLC method to determin the content.3.Used Oxford cup method,double dilution method and eliasa determination OD method to preliminarily study on in vitro antibacterial activity of the root bark of 5%sulfuric acid extracts,and its 95%ethanol,70%ethanol,distilled water pats and these parts of the root xylem and monomer compounds to common pathogens and probiotics.Results:1.?1?Berberis Heteropoda root bark and xylem contained many kinds of compounds.The content of total alkaloids was determined by UV?average 2.94%?was higher than that of acid dye Colorimetry?average1.42%?.The total alkaloids content of different solvent extracts of root bark and root xylem was determined by UV?root bark/root xylem?:acid extract?4.52%/3.96%?>ethanol extract?2.89%/2.88%?>water extract?1.74%/2.28%?>acetone extract?0.83%/1.10%?>ethyl acetate extract?0.28%/0.13%?>petroleum ether extract?0.07%/0.08%?.The optimum extraction condition of total alkaloids was extracted 2h with 51 times 50%ethanol at80?,three extraction cycles.Through verification test,the total alkaloids extraction rate was 4.01%,RSD=1.12%,indicating that the optimized process was stable and feasible.2.The 5%sulfuric acid extracts were isolated and purified to obtain the compounds,which was identified as trans-ferulic acid.And the content was 0.59%3.Root bark of 5%sulfuric acid and its 95%ethanol parts and the same parts of root xylem had inhibitory effect on four kinds of pathogenic bacteria.The MIC of root bark of 5%sulfuric acid extract to the pathogenic bacteria was 2.02mg·mL-1,and its 95%ethanol part MIC to Escherichia coli was 2.13mg·mL-1,to Enterococcus faecalis was 4.25mg·mL-1,to Pseudomonas aeruginosa was 8.50mg·mL-1,to Staphylococcus aureus was 1.06mg·mL-1;The MIC of root xylem 5%sulfuric acid extract on the four kinds of pathogenic bacteria were 1.90mg·mL-1,and its 95%ethanol part MIC to Escherichia coli and Enterococcus faecalis were 5.05mg·mL-1,to Pseudomonas aeruginosa was 20.19mg·mL-1,to Staphylococcus aureus was 2.53mg·mL-1.The water part(20.68-10.34mg·mL-1)of 5%sulfuric acid extract of root bar,and 95%ethanol part(2.53-10.10mg·mL-1)and water part(0.97-7.73mg·mL-1)of 5%sulfuric acid extract of root xylem had a promoting effect on the growth of Lactobacillus;The 70%ethanol part(40.06mg·mL-1)of 5%sulfuric acid extract of root xylem,the water part(20.68mg·mL-1)and 70%ethanol part(2.91-11.63mg·mL-1)of 5%sulfuric acid extract of root bar had a promoting effect on the growth of bifidobacteria,the remaining samples showed inhibitory effect on the two kinds of bacteria.Conclusion:1.?1?The chemical composition of root bar and root xylem were almost the same.The alkaloid content was high,and if the alkaloid is used as the active ingredient,root xylem can also be used as the drug site.?2?The optimized extraction process of total alkaloids in root bark was stable and simple,and could provide experimental basis for follow-up study.2.The 5%sulfuric acid extract of root bark contains trans-ferulic acid,and the content was 0.59%.3.The 5%sulfuric acid extract of root bark and xylem on Staphylococcus aureus,Escherichia coli,Pseudomonas aeruginosa and Enterococcus had different degrees of inhibition,a proliferation promoting effect on Bifidobacterium and lactobacillus.The preliminary inference,if used in the treatment of intestinal disease caused by bacterial flora disorder,Berberis Heteropoda root xylem also can be used in medication site. |
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