知母化学组分特征及其影响因素研究

【作者】 孙小明；

【导师】 王文全；

【作者基本信息】 北京中医药大学 ， 中药学， 2008， 硕士

【摘要】 知母始载于《神农本草经》,列为中品,是百合科Liliaceae知母属植物知母Anemarrhena asphodeloides Bge.的干燥地下根茎。是临床常用重要中药,具有清热泻火、生津润燥的功效,常用于外感热病、高热烦渴、肺热燥咳、骨蒸潮热、内热消渴、肠燥便秘等证。主要分布于我国北方大部分地区,不同产地药材中各化学成分含量及比例关系也存在较大差异,因此是导致知母药材质量不稳定的主要原因之一。迄今为止,知母药材质量评价研究仍主要针对少数几个成分,不能较全面的反映知母化学组分特征。随着野生知母日益减少,市场中栽培知母比例升高,同时栽培措施不统一,药材加工不规范,也是药材质量不稳定的一个主要原因。本文针对以上问题,在现有文献资料基础上,开展了知母药材调查收集工作,通过田间试验和实验室分析,探索了不同产地野生和栽培知母的化学组分及微量元素的特征,研究气候因子与化学组分间的相互关系及栽培措施对知母中化学组分含量的影响。(1)建立了HPLC-ELSD知母指纹图谱。对新芒果苷、芒果苷、知母皂苷C、知母皂苷AⅢ主要成分进行了指认。15个产地野生和9个栽培产地知母图谱相似度均在0.97以上。不同产地野生和栽培知母间在两个特征区域存在差异。(2)全面分析了化学组分与气候因子的相关性。有效成分与6种气候因子均相关不显著(P>0.05)。粗蛋白与7月均温和年降水量呈显著性相关(P<0.05)。总多糖与年降水量、无霜期呈显著相关(P<0.05),与7月均温呈极显著相关(P<0.01)。(3)探讨了栽培措施对化学组分的影响。施肥对知母的有效成分含量和产量有很大的影响;移栽密度对有效成分影响不显著。使用适当配比的N、P、K肥,对增加产量起着重要的作用。(4)揭示了知母微量元素的含量特征,探讨了环境和栽培措施对其含量的影响。不同产地野生知母中Mn、Fe、P元素含量差异显著(P<0.05),不同产地栽培知母中Fe、P、Mg、Na、Si元素含量差异显著(P<0.05)。Mn元素与蒸发量呈现极显著正相关(P<0.01),与年降水量成正相关。不同生长年限知母中Na元素含量存在显著差异(P<0.05),四年生知母的Na含量较高。喷钾肥试验的各处理间K含量存在极显著差异(P<0.01),一年喷三次有利于钾元素的积累,从而促进知母根茎的生长。(5)初步估算了知母中主要化学组分的含量范围。粗脂肪含量范围在1.27～1.71%,粗蛋白含量范围在6.91~9.42%,总多糖含量范围在13.8~18.4%,菝葜皂苷元含量范围在0.87~1.89%,知母皂苷C含量范围在1.05~1.97%,知母皂苷AⅢ含量范围在0.34~0.99%,新芒果苷含量范围在0.80~1.80%,芒果苷含量范围在0.45~1.15%。本文的特色有以下几个方面:(1)建立了知母粗脂肪、粗蛋白的提取测定方法。(2)建立了知母HPLC-ELSD指纹图谱,对新芒果苷、芒果苷、知母皂苷C、知母皂苷AⅢ四种成分进行了指认。(3)从药材微量元素的角度揭示了知母的质量特征。更多还原

【Abstract】 Common Anemarrhena Rhizome was firstly recorded on the“Shen Nong Ben Cao Jing”, which was listed as a middle one. It is the dry rhizome of Anemarrhena asphodeloides Bge., belonging to Liliaceae. It is commonly used on clinic as the heat-clearing and fire-purging drug and is widely distributed in the North China, Korea and Mongolia. The quality is different because there are many resouses, such as different places of products and collection periods. The quality study of it was mainly concerned on several constituents, such as sarsasapogenin and mangiferin. The whole quality character couldn’t be reflected by these analyses.Meanwile, there was no standard of cultivation technique when producing. So the quality of it is difficult to guarantee.This article is mainly to solve the above problems and analyse quite a lot of samples which were collected in the North China by wild investigation, field experiment and laboratory analysis. In order to identify the quality character and the influencing factors on the contents and proportion of constituents, and the relationship between the contents and the climatic factors, we do the research on the quality character of the wild growth and the cultivated ones by studing the multiple constituents determination and HPLC fingerprint.(1)HPLC–ELSD fingerprint was set up. The four contents neo-mangigein, magigein, timosaponin C, timosasponin AⅢare identified.The semblance of the two fingerprint are all above 0.97. There are some differences between two regions.(2)The relationship between the contents and climatic factors was analysed. It shows that the sarsasapogenin, mangigerin, neo-mangigein, timosaponin C timosasponin AⅢand crude fat all have not significant difference(P>0.05)with year sam, July sam, annual precipitation, evaporation capacity, sunshine duration, frost-free period. The crude protein has significant difference(P<0.05)with July sam and annual preipitation. The polycose has significant difference(P<0.05)with annual preipitation and frost-free period, and has most significant difference(P<0.01)with July sam.(3)Investigate the influnce of cultivate measures to constituents. the compound fertilizer and farmyard manure have significant influence to the active contents.There is no significant influence to the active contents in different margins. The content of crude fat is significant different(P<0.05) under different manure match. Rational use N,P,K and proper match are good for accumulation of crude fat and protein and also for increasing the outcome.(4)Reveal the content character of microelements and investigate the influence of enviroment and cultivated measure to the content of constituents. The content of Mn has significant direct correlation(P<0.01)with evaporation capacity. The content of Mn has inverse correlation with year sam, July sam, sunshine duration and frost-free period. The content of P has no significant relationship with the six factors(P>0.05).It has direct correlation with evaporation capacity and inverse correlation with the others.The content of Na+ with different growth time limit are significant different(P<0.05),and the content is highest of the fourth age. The content of K+ are significant differen(tP<0.01) between each handling in potash manure. It is good for the accumulation of K when consperg three times each year.(5)Estimate the range of content of chemical constituent. The content range of the crude fat, crude protein, total polysaccharide, sarsasapogenin, Anemasaponin C, AnemasaponinAⅢ, neomangiferin, mangiferin are respectively 1.27～1.71%, 6.91~9.42%, 13.8~18.4%, 0.87~1.89%, 1.05~1.97%, 0.34~0.99%, 0.80~1.80%, 0.45~1.15%.The distinctness of this article have the following aspects:(1) The methods of extracting contents of crude fat and crude protein were set up.(2) The HPLC-ELSD fingerprint was set up. The four active compounds were identified.(3)Determine the contents of ten kinds of microelements. Reveal the quality character from the angle of microelements.更多还原