HPLC Analysis Of Several Kinds Of Polyphenols Of Bergenia Scopulosa T.P. Wang And Rodgeris Aesculifolia Batal

Bergenia scopulosa T.P.Wang and Rodgeris aesculifolia Batal are two ofChinese traditional drugs of Saxifraceae plants with the function of removing weaknessand cough. Polyphenol compoudes are the main components in these two kinds ofplants. About the determination of chemical components, there were many reports ondetermination of bergenin, but there are no report on simultaneously determination ofbergenin, catechin, arbutin and gallic acid with HPLC.There are two parts including summary and research in the thesis:Part one: Review1. Including present situation of chemical ingredient research and pharmacologyresearch of Bergenia scopulosa T. P. Wang and Rodgersia aesculifolia Batal. There arepolyphenols, yellow ketones, anthracene quinone type, organic acids and otherinorganic substance in these two kinds of plants. Their pharmacology activeness arewidespread.2. Including research progress of polyphenol pharmacology and the method ofdetermination; and research progress of pharmacology activeness and the determinationmethod of highly effective liquid chromatography of bergenin, catechin, arbutin andgallic acid.Part two: Research papersIn this part, two departs are involved.1. The optimum process of extracting total polyphenol from the root of Rodgersiaaesculifolia Batal by using univariant method was studied and optimized withorthogonal test, then the content of total polyphenol was monitored by colorimetry ofdiazotization reagent at 415 nm. Experiments were made by using L₉(3⁴) orthogonaldesign with extractive rate of total polyphenol as the index, investigating the influenceof extracting solvent, the concentration of solvent, the ratio of solid to liquid andextracting time. The obtained optimum extracting condition of total polyphenol is thatthe material was refluxed in 10 times (m/v) 60% of neutral methanol solution (v/v) for30min with 3 times. The content of total polyphenol from Rodgersia aesculifolia Batalwas 28.32%. The results indicated that the optimum extraction conditions of total polyphenol was rational.2. A direct, reliable and accurate high performance liquid chromatography (HPLC)method has been developed using danshensu as an internal standard for thequantification of gallic acid, arbutin, bergenin and catechin in Bergenia scopulosaT. P. Wang and Rodgersia aesculifolia Batal, two important traditional Chinesemedicines. The separation of these components was achieved on a C₁₈ column withgradient elution using acetonitrile-methanol (80/20, v/v) and water (containing 0.03M/Lpotassium dihydrogen phosphate adjusted to pH 3.50 with phosphoric acid) as mobilephase, with a gradient elution of 8-8%(v/v) A at 0-10min, 8-25%(v/v) A at 10-30 min,25-25%(v/v) A at 30-50min.. The solutions were chromatographed at constant flow rateof 0.5ml/min and UV detection was set at 230nm. All calibration curves showed goodlinearity (r²ï¼ž0.9982) with test ranges. Intra-assay precision of four components wereless than 1.4% and the average recovery rates obtained were in the range of95.66—104.9% for all with R.S.D below 2.9%. Method was validated according to ICHguidelines and proved to be suitable for stability, homogeneity testing and qualitycontrol of these compounds in pharmaceutical applications.Determining under the designation chromatograph conditions, the chromatographpeak shape and the degree of dissociation of the four ingredients quite are all satisfied.The method may be used to determine the analogous compound. And we have allcarried on the determination of these four kinds of chemical ingredients of the differentspot of the two plants, which may be used for these two kinds of plants' quality control.This experiment research the optimum process of extracting total polyphenol ofRodgersia aesculifolia Batal for the first time and firstly establish the method for thedetermination of the total polyphenol of arbutin, gallic acid, bergenin and catechin ofthe two medical plants simultaneously. And at the same time, arbutin, gallic acid,bergenin and catechin of the different parts of the two plants has carried on thedetermination for the first time by the established method. These have established thefoundation for the further research and development of Bergenia scopulosa T. P. Wangand Rodgersia aesculifolia Batal.