| Anoectochilus roxburghi, which is belongs to genus Cypripedium of the orchidfamily, is one kind of perennial herbs. Anoectochilus roxburghi, which is one of thetraditional medicinal materials in our country, has multi-functions like heat-clearingand detoxifying, nourishing blood and reduce internal heat, anti-inflammatory andanalgesic effects. It has a good clinical efficacy on sore throat, fever, Antidiarrheal,snake wound. Anoectochilus roxburghi is safe and simple with a little side effect. Itcould be used for treating these diseases, including diabetes, hyperlipidemia, tumorand hepatitis B in China. As the research moves along, Anoectochilus roxburghi’sgood quality become more prominent and more favorite and has avast development future. Based on the conditions of widespread, versatility andcurative effect, we study mainly effective material basis on hypolipidemic ofAnoectochilus roxburghi. The extracting and purifying process of kinsenoside fromAnoectochilus roxburghi and the determination method for the main content ofkinsenoside have been investigated.Study of preparing technologyAccording to the consult literature material determined this display curativeeffect the main active constituent, it has determined that it is glycosides includingkinsenoside. We carry on the analysis using the statistical software to each tentativedata, thus optimal related experimental parameter.Extraction technological processBecause of the larger polar character of kinsenoside, it is soluble in water anddissolves in ethyl alcohol.So we investigate the factors of extracting solvent to findthe best procedure. Because of the thermal instability of kinsenoside, itis easy to decompose.So we investigate the factors of extraction temperature to findthe best procedure.Extract research: Methods: To optimize the extraction process of Anoectochilus roxburghi by orthogonal test. Based on the amount of kinsenoside, we studied the extraction temperature, extraction time, extraction times and liquid-solid ratio, at last obtained the best extraction conditions. Results:The optimum technical condition is:adding15times amount of water, temperature is80℃, extracting3times and every time with2hour.Purification technology processAlcohol precipitation research: Because Anoectochilus roxburghi contains aconsiderable amount of sugars, we need remove the saccharide impurities by alcoholprecipitation. Methods: Based on the amount of impurities, we studied the ethanolconcentration, at last obtained the best alcohol precipitation conditions. Results:The optimum technical condition is: extract enriched to relative density is1.13（45℃）, by adding ethanol containing alcohol volume is80%, aside12hours aftermixing.Column refining research: Methods: In order to remove the saccharide impurities and flavone impurities, we studied the resin type, at last obtained the best column refining conditions. Results: The optimum technical condition is: using the UBK555ion exchange resin to refine the Anoectochilus roxburghi extract.Study of quality controlMethods: HPLC-ELSD was used to determine the content of kinsenoside. Thegradient elution was performed on a Hypersil NH2（5μm，4.6×250mm）column withacetonitrile: purified water（92:8）as the mobile phase. The flow rate is1.0ml/min.The column temperature is30℃. The ELSD spray chamber temperature is40℃.Thenitrogen pressure is350kPa. The gain is seven. Results: The calibration curves（theIn values of peak areas with corresponding In values of sample introducingamounts）were linear within the range of0.524~5.24μg for kinsenoside, the averagerecovery rate was98.22%, RSD=1.33%. Conclusion: The determination method issimple, reliable, accurate and reproducible. It is suitable for the quality control ofAnoectochilus roxburghi effective-part. |
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