【作者】 陈佳江；

【导师】 董小萍；

【作者基本信息】 成都中医药大学 ， 中药学， 2013， 博士

【摘要】 目的：本课题在科技部“十二五”支撑项目——石斛规范化种植基地及其SOP优化、升级研究(2011BAll3B02-8)的资助下,对川产道地药材叠鞘石斛与金钗石斛、鼓槌石斛、流苏石斛之间及叠鞘石斛不同部位(茎、叶、花)化学成分指纹图谱及含量进行对比研究,以期发现叠鞘石斛与三者间的生物亲缘关系及对叠鞘石斛的综合利用提供参考：并对叠鞘石斛化学成分进行分离鉴定,阐明叠鞘石斛的化学成分类型,为其质量控制提供一定的依据；并对叠鞘石斛进行配方颗粒的制备研究,扩大其应用范围,增加种植者的经济收入,提高种植积极性,进一步带来更大的效益,为石斛资源的可持续利用和合理开发奠定基础。方法：采用薄层色谱法、紫外-可见分光光度法、高效液相色谱法、气相色谱等现代分析技术,从“薄层色谱——高效液相色谱——气相色谱——单体成分／组分含量”四个方面,对叠鞘石斛与金钗石斛、鼓槌石斛、流苏石斛及叠鞘石斛茎、叶、花化学成分进行对比研究；采用现代色谱分离方法及分析手段,对叠鞘石斛化学成分进行分离鉴定,并对其进行含量测定；从提取、干燥工艺,辅料选择,成型工艺等方面对叠鞘石斛配方颗粒进行制备工艺研究。结果：四种石斛对比叠鞘石斛中含有石斛酚,但未检出石斛碱和毛兰素。TLC与HPLC结果互为补充,直观显示四种石斛在化成成分上的异同点。脂溶性成分GC-MS对比分析结果对其具有一定的相同成分作出了验证。相同色谱信息表明具有同化学成分,作为同属植物其可能具有相类似的功效；特异性色谱信息表明其在化学成分上有所差异,在具体功效主治上是否完全相同应加以重视；对比结果提示叠鞘石斛及流苏石斛化学成分的研究具有较大的潜力。含量测定结果显示,在糖类成分含量上金钗石斛与流苏石斛较为接近,鼓槌石斛与叠鞘石斛较为接近,推测其在调节机体免疫、抗肿瘤及调节血糖等方面具有类似药效,但在使用剂量上应有所不同。四种石斛中石斛酚含量差异较大,流苏石斛与叠鞘石斛中含量较高约为220gg／g,提示其在抗糖尿病性白内障方面具有一定的研究价值。通过游离糖类衍生化后GC-MS分析结果可知,四种石斛中均主要检出鼠李糖、阿拉伯糖、甘露糖和葡萄糖,均未检出乳糖,金钗石斛亦检出含有半乳糖。叠鞘石斛茎、叶、花对比TLC对比结果显示,叠鞘石斛茎的色谱斑点较为丰富；HPLC对比结果显示,叶和花色谱峰信息较为丰富,且与茎中化学成分具有一定的相似性,推测其可能具有相同的化学成分及类似功效。GC-MS结果显示叠鞘石斛茎、叶、花有4种共有成分,且茎与叶、花分别有8种、13种共有成分；茎中烷烃类成分含量较高,而叶和花中脂肪酸、酯类含量较高,且成分种类明显多于茎：提示叠鞘石斛叶化学成分具有一定的研究意义,可为其综合利用提供依据。糖类成分含量测定结果表明,茎中除还原糖含量明显低于叶外,多糖及水溶性总糖的含量均高于叶中含量。测定结果提示可对叠鞘石斛叶在多糖所具的药理活性方面具有一定的研究价值。叠鞘石斛化学成分分离鉴定及含量测定对叠鞘石斛乙酸乙酯部位化学成分进行分离纯化,共鉴定出17个化合物,其中4,5-二羟基-2-甲氧基-9,10-二氢菲、丁香酸、大黄素、松柏醛、香兰素、邻苯二甲酸二(2-乙基)己酯、5-乙酰氧甲基糠醛、丁香醛、5-羟甲基糠醛、香草酸均为首次从叠鞘石斛中分离。采用HPLC法对叠鞘石斛中4,5-二羟基-2-甲氧基-9,10-二氢菲含量进行测定,结果表明其含量为17.42μg／g,其含量测定结果为叠鞘石斛的质量控制提供了一定的参考。4,5-二羟基-2-甲氧基-9,10-二氢菲及柚皮素分离纯化,为叠鞘石斛具有抗癌活性的药理作用提供了一定的依据；石斛酚与丁香酸的分离纯化,为叠鞘石斛在抗糖尿病性白内障方面可能具有药理活性提供了一定的参考。叠鞘石斛配方颗粒制备工艺研究通过实验研究,确定了叠鞘石斛配方颗粒的制备工艺；通过中试研究,证明该工艺可用于工业化生产且稳定可行；对中试样品进行含量测定,其指标性成分转移率达82.8%,表明该工艺符合工业化生产要求。该配方颗粒的研制,为扩大叠鞘石斛市场范围及其后续产品开发奠定了基础。结论：通过对比研究,为叠鞘石斛的药用价值及综合利用提供了一定的参考;化学成分的分离为叠鞘石斛的物质基础及药理活性提供了一定的支持;含量测定结果为叠鞘石斛的质量控制提供了一定的依据；配方颗粒的制备研究,为扩大叠鞘石斛市场应用及后续产品开发奠定了基础。更多还原

【Abstract】 Objective:Comparative studie between D.aurantiacum Rchb. f.var.denneanum (Kerr) Z.H.Tsi, D.nobile Lindl., D.chrysotoxum Lindl., and D.fimbriatum Hook.. Also investigate the stem, leaf, flower of D.aurantiacum Rchb. f.var.denneanum (Kerr) Z.H.Tsi. To found the biological genetic relationship of D.aurantiacum Rchb. f.var.denneanum (Kerr) Z.H.Tsi and those species, and provide the reference for comprehensive utilization of D.aurantiacum Rchb.; Isolation and identification of the chemical composition of the D.aurantiacum Rchb., to clarify the type of chemical composition in it and to provide a basis for its quality control. To investigate preparation process of formula particles of D.aurantiacum Rchb. to expand the range of applications and increase the income of the growers, improve planting initiative, to bring greater economic benefits.To laid the foundation for the sustainable use and rational development of D.aurantiacum Rchb.Method:The use of TLC, UV-Vis, HPLC, GC and other modern analytical techniques, from Four aspects of different levels to investigate the chemicalc onstituents of4species of Dendrobiumand the stem, leaf, flower of D.aurantiacum.Rchb. To isolated and identified the chemical constituents of D.aurantiacum.Rchb., and to determine the content of it. To investigate preparation process of formula particles of D.aurantiacum Rchb. from the extraction, drying process, materials selection, molding process, Etc.Result:contrast research of Four DendrobiumD.aurantiacum Rchb.contains gigantol, but not the detection Dendrobine and Erianin. The detectable result of TLC and HPLC complement each other, and visualize the similarities and differences of chemical composition of four dendrobium.the comparative analysis result of fat-soluble ingredients by GC-MS has verified that four Dendrobium with the same ingredients. The same chromatographic information to show that they have the same chemical composition, as the same plant that may have a similar effect. Specific chromatography show that the differences in the chemical composition,and should be pay attention that whether they have the the same functions and indications. Comparison results prompte that the chemical constituents study of D.aurantiacum Rchb. and D.fimbriatum Hook.having a greater potential.The result of Content Determination show that D.nobile Lindl. and D.fimbriatum Hook.have similar content of carbohydate components, while D.chrysotoxum Lindl. and D.aurantiacum Rchb. are similar., speculated to have similar efficacy in immune regulation, anti-tumor and regulation of blood sugar, but the dose should be different. The content of gigantol is quite different in different Dendrobium, D.aurantiacum.Rchb. and D.fimbriatum Hook.has the high content about220μg/g, suggesting that it has a certain value in the anti-diabetic cataract.Test results of free sugars by GC-MS show that rhamnose, arabinose. mannose and glucose were detected, but not lactose. D.nobile Lindl. also found to contain galactose.Comparative study of the stem, leaf, flower of D.aurantiacum Rchb. f.var.denneanum (Kerr) Z.H.Tsi.The comparison results of TLC show that the chromatography spots of D.aurantiacum.Rchb. are abundant. HPLC comparison results show that the leaf and flower have more abundant of chromatographic information, speculated that it may have the same chemical composition and similar efficacy. The results of GC-MS show that there are four common components in stem, leaf and flower. Stem have8.13common components with leaf and flower. Stem has higher content of alkanes, but leaf and flower with higher levels of fatty acids and esters, and the component types of leaf and flower significantly more than the stem.Prompt that chemical composition in leaf has a certain significance, and can provide the basis for its comprehensive utilization.The the content determination of carbohydrate show that the content of reducing sugar in stem is lower than in leaf, but the other are higher than it. The determination results suggest that leaf could has a certain research value about pharmacological activity of polysaccharideIsolation and identification of chemical composition and content determination of D.aurantiacum.Rchb.To separation and purification the chemical composition of D.aurantiacum.Rchb.. a total of17compounds were identified. Among those, coniferaldehyde, vanillic acid, vanillin, bis(2-ethylhexyl)phthalate, syringaldehyde,5-hydromethylfuraldehyde,4,5-dihydroxy-2-methoxy-9,10-dihydrophenanthrene,5-acetoxymethy lfuraldehyde, syringic acid and andemodin are first separated from D.aurantiacum Rchb.The content determination result by HPLC of4,5-dihydroxy-2-methoxy-9,10-di-hydrophenanthrene in D.aurantiacum.Rchb. show that the content is only17.42μg/g. The result of content determination to provide a reference of quality control of D.aurantiacum.Rchb. The separation and purification of4,5-dihydroxy-2-methoxy-9,10-dihydrophenanthrene and naringenin to provide some basis of anti-cancer activity of D.aurantiacum.Rchb. The separation and purification of gigantol and syringic acid to providing a reference that D.aurantiacum.Rchb. might have pharmacological activity in the anti-diabetic cataract.Investigate preparation process of formula particles of D.aurantiacum Rchb.Through experimental studies to determine the preparation process of formula particles, the preparation process is stable, viable, and can be used for industrial production. The content determination result showed that the transfer rate of target component is82.8%, indicating that the preparation process to meet the requirements of industrial production, to expand the scope of the market and laid the foundation for the rational development of D.aurantiacum Rchb.Conclusion:Through comparative studies to provide some reference for medicinal value and utilization of D.aurantiacum Rchb. f.var.denneanum (Kerr) Z.H.Tsi. The separation of chemical composition to provide some support the material foundation and pharmacological activity of it. The content determination provide some basis for quality control of it. The preparation technology of formula particles to expand the application of the market and laid the foundation for the rational development of it.更多还原