龙胆的质量评价
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摘要
中药龙胆为龙胆科(Gemianaceae)植物条叶龙胆(Gentiana manshurica Kitag.)、龙胆(Gentiana scabra Bge.)、三花龙胆(Gentiana triflora pall.)或坚龙胆(Gentiana rigescens Franch.)的干燥根及根茎。具有清热燥湿,泻肝胆火的功效,用于治疗湿热黄疸,阴肿阴痒、带下、强中、湿疹瘙痒、目赤、耳聋、胁痛、口苦、惊风抽搐等症状。
     本实验共分六章,从灰分、水分、醇浸物、多糖、龙胆苦苷的含量上,对鸡西等10个不同产地、左家龙胆GAP示范基地的10个不同采收期的龙胆进行了较为系统的比较,并对龙胆多糖、龙胆挥发油的提取工艺进行了研究,对四个不同产地龙胆的药理作用进行了比较分析。
     10个不同产地和GAP示范基地的10个不同采收期龙胆的灰分含量均达到2005年版《中国药典》的规定标准。10个不同产地龙胆水分含量在6.9401%~5.3648%;醇浸物含量在41.1780%~15.3407%之间,差异显著。10个不同采收期龙胆水分含量在9.6281%~5.0488%;醇浸物含量在40.8272%~33.8837%。其余均在此范围内波动。
     龙胆多糖的研究主要采用正交试验的方法对超声波法及微波法提取龙胆多糖的工艺进行研究,分别确定超声波法及微波法提取龙胆多糖的最佳提取工艺,并与传统的水煎煮法进行比较,选择出三者中的最佳提取方法。并利用所选定的最佳工艺对10个不同产地、10个不同采收期的龙胆的多糖含量进行提取、测定。确定了超声波提取龙胆多糖的最佳工艺条件为超声温度为80℃,提取时间为20min,溶剂量为8倍;微波提取龙胆多糖的最佳工艺条件为微波功率为450w,溶剂量为14倍量,提取时间为4min。且超声波法提取龙胆多糖优于微波法,二者均优于传统的水煎煮法。各个不同产地龙胆多糖含量差别较大,其中以安徽省桃花潭地区龙胆的龙胆多糖含量最高为24.8747%,黑龙江省鸡西市的最低为4.6989%。各不同采收期的多糖含量从8.1516%到24.5675%。这为大量工业化提取龙胆多糖奠定了基础,也为进一步研究龙胆多糖的其药理作用提供了物质前提。
     龙胆苦苷的研究是采用VPODS柱(150mm×4.6mm,0.25μm),按照药典规定的龙胆苦苷的测定方法,对10个不同产地、10个不同采收期的龙胆中龙胆苦苷的含量进行测定,应用DPS数据处理系统对其进行分析,并对东北三省省间数据进行处理。结果发现不同产地的龙胆以侯马产地的龙胆,龙胆苦苷含量最低,为0.5136%,未达到2005年版《中国药典》的标准(不得少于1.0%),以左家地区龙胆中的龙胆苦苷含量最多,为9.8945%,其余产地龙胆的龙胆苦苷含量均在此范围内波动,均达到2005年版《中国药典》的规定值,其问相互差异极显著。各个采收期龙胆中龙胆苦苷含量均达到标准,在3.3528%~7.2726%范围内波动,并具有显著差异。
     龙胆挥发油的研究是采用CO_2超临界流体萃取法提取龙胆挥发油类成分,建立CO_2新的提取方法-超临界流体萃取法提取龙胆挥发油的最佳提取工艺。并利用GC-MS对CO_2超临界流体萃取法及传统的水蒸气蒸馏法提取龙胆中挥发油类成分进行了比较分析。结果建立CO_2超临界流体萃取法提取龙胆挥发油的最佳工艺条件为萃取釜温度为30℃,样品粒度为20~40目之间,萃取压力为25MPa。共鉴定出50个成分,其中CO_2超临界萃取物被鉴定了28个成分,水蒸气蒸馏法萃取物被鉴定了27个成分,两者共有成分为5个。CO_2超临界流体萃取法的提取龙胆挥发油无论从提取物提取时间、颜色、得率、化合物成分及含量等方面均优于水蒸气蒸馏法。
     以Wistar大鼠为实验动物,建立CCl_4致大鼠慢性肝损模型,以龙胆治疗肝损伤疗效为指标,对桓仁、左家、新宾、绥化4个不同产地的龙胆在治疗CCl_4致大鼠慢性肝损伤疗效的差异进行了比较分析,探索其药理作用差异的原因。结果发现4个产地总体药理作用左家龙胆最佳。
     本文针对不同产地、不同采收期龙胆灰分限量、水分限量、醇浸物含量以及龙胆的多糖的提取工艺、挥发油的不同产地、不同采收期的龙胆苦苷化学成分、药理作用做出比较研究,为其质量标准的建立奠定了良好的基础。为龙胆全面的质量标准的建立奠定了基础。
Gentian is the roots of plants Gentiana manshurica Kitag.、Gentiana scabra Bge.、Gentiana triflora pall. or Gentiana rigescens Franch. of Gentianaceae. It has the efficacies of clearing heat and drying damp,discharging fire for liver and gall and damp-heat jaundice,hidden-arsine gargalesthesia, morbid leucorrhea, persistent erection, eczema pruritus, conjunctival congestion, deafness, hypochondriac pain, bitter taste, infantile convulsion for therapies
     This experiment is divided into six chapters altogether, including systemic comparison for ash contents, water contents, alcohol extractive contents, polysaccharide contents and gentiopicroside contents in gentian roots from different areas or harvesting in different times, studies on extraction technology for polysaccharide and essential oil from gentian roots, comparison and analysis of the pharmaceutical action of alcohol extractive in gentian roots from four different areas.
     Samples from 10 producing areas in 6 provinces altogether were collected for experiment analyzing. The samples were gentiana roots from Gentiana scabra Bge., Gentiana manshurica Kitag. The 10 different times Gentiana scabra Bge. Samples were collected from the beginning of September to the end of October in Zuo Jia in 2005.
     The ash contents in gentian roots from different areas or harvesting in different times achieve the pharmacopeia ordain cost. The water contents in gentian roots from different areas were from 5.3648% to 6.9401%, and 5.0488% to 9.6281% in different times. The alcohol extractive contents in gentian roots from different areas were from 15.3407% to 41.1780%, and 33.8837% to 40.8272% in different times.
     The chapter of study for gentian polysaccharide is to optimize and establish the extraction process of gentian polysaccharide. To extract gentian polysaccharide and determine contents in gentian roots from different areas or harvesting in different times. The ultrasonic wave extraction optimum process parameters are as fouons: temperature is 80℃, extract time is 20m, mass ratio of water is 8 times. The microwave-assisted extraction optimum process parameters are as fouons: wave power is 450 w, mass ratio of water is 14 times, extract time is 4m. The ultrasonic wave extraction is better than microwave-assisted extraction for gentian polysaccharide extraction, and they are both better than water decoction. The difference is marked of polysaccharide contents in different areas. The hightest polysaccharide content is 24.8747% in Tao Hua-tan of An Hui Pronvince, the lest is 4.6989% in Ji Xi of Hei Long-jiang Province. The contents are from 8.1516% to 24.5675% in different times. This study establish foundation for gentian polysaccharide extraction by industrialization, and afford material antecedent for further research into pharmacodynamic function of the polysaccharide in gentian roots.
     The chapter of study for gentian gentiopicroside is to determine the contents of gentiopicroside in gentian roots from different areas or harvesting in different times, according to method in pharmacopeiae, used VPODS(150mm×4.6mm, 0.25μm). To analysis by the data processing, found that the lest content is 0.5136% in Hou Ma of Shan Xi Province, don not achieve the pharmacopeia ordain cost. The highest content is 9.8945% in Zuo Jia of Ji Lin Province. The contents are from 3.3528% to 7.2726% in different times.
     The chapter of study for gentian essential oil is to establish the best extraction by SFE-CO_2. Use GC-MS to compare the essential oils extracted from Radix Gentianae by supercritical fluid extraction(SFE) and steam distillation(SD). The SFE-CO_2 optimum process parameters are as fouons: the temperatrue is 30℃, the garticle size is 20~40 mesh, the extraction compressive stress. There were 28 components from Radix Gentianae by SFE were separated and identified. There were 27 components from Radix Gentianae by SD were separated and identified. The SFE-CO_2 is better than the steam distillation.
     Establishment of experimental model for liver injury of CCl_4 in Wistar rats. Compare the therapies of alcohol extractive in gentian roots from four different areas. Result is that Zuo Jia is the best.
     This essay aim at ash contents, water contents, alcohol extractive contents, polysaccharide contents and gentiopicroside contents in gentian roots from different areas or harvesting in different times, and study on the extraction for polysaccharide and essential oil in gentian roots. All is to establish the good foundation for the quality standardization of gentian.
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