【作者】 耿志鹏；

【导师】 张艺； 瞿显友；

【作者基本信息】 成都中医药大学 ， 药物化学， 2010， 硕士

【摘要】 黄连为毛莨科植物黄连(Coptis chinensis Franch.)、三角叶黄连(C.deltoidea C.Y.Cheng et Hsiao)、云南黄连(C.teeta Wall.)的干燥根茎。药材依次习称“味连”、“雅连”、“云连”,始载于《神农本草经》。味连也称鸡爪连、川连,为目前应用最广的黄连品种。《唐本草》载：“蜀道者粗大,味极苦,疗渴为最,江东者节如连珠,疗痢大著。”,李时珍谓：“今虽吴、蜀皆有,惟以雅州、眉州者为良。”。不同产地黄连的化学成分受其不同环境的影响而可能表现出差异性,其中川产黄连因其特有的遗传背景、生态环境、文化背景及中医药理论而具有道地性。目的：开展黄连的化学分析及生态因子研究,并分析两者间的关系,以此,建立反映川产黄连药材道地性的质量评价方法。方法：1、收集黄连产区的药材样品,考察产区的气候因子,并收集各产区的土壤样品,测定土壤样品的pH、有机质、水解性氮、速效磷、速效钾的含量,对黄连不同产区生态因子的差异性进行分析。2、采用NIR建立不同产区黄连样品的NIR指纹图谱,对其进行主成分分析,研究不同产地样品的差异性。3、采用RP-HPLC法建立不同产地黄连样品的HPLC指纹图谱,测定其中的盐酸药根碱、盐酸非洲防己碱、盐酸表小檗碱、盐酸黄连碱、盐酸巴马汀和盐酸小檗碱等6种主要生物碱含量。采用MPLS将HPLC结果与NIR数据进行回归,建立NIR测定黄连中6种生物碱的模型方程,对预测集中各成分的NIR预测值与HPLC真值间进行配对数据双尾T检验。4、运用聚类分析、灰色关联和典型相关分析等数理统计学方法,分析黄连产地生态因子与其化学成分之间的相关性。结果：1、收集到不同产地黄连药材样品134份及土壤样品19份。土壤分析聚类结果表明,重庆石柱地区与湖北利川地区的土壤样品具有相似性,而四川产地的土壤样品也相类似。2、NIR指纹图谱主成分分析结果表明,对于采集于不同产地五年生样品及不同产地的加工品,四川产地味连和重庆与湖北的样品具有明显差异；而且散点图可以将四年和七年样品与五年的样品分开。3、检测了27批样品的HPLC指纹图谱,共标定12个共有峰,相似度大于0.99。聚类结果表明,重庆石柱、湖北利川与湖南样品比较相似；峨眉山市、洪雅县样品有相似性；彭州、崇州及邛崃市样品则大多独自聚在一起,其余少数与峨眉山市及洪雅县样品聚在一起。4、建立了HPLC测定6种生物碱含量的方法,各生物碱的平均加样回收率依次为101.57%(RSD=1.33%),102.50%(RSD=1.35%),100.80%(RSD=1.91%), 102.66%(RSD=1.23%),97.85%(RSD=1.25%),99.01%(RSD=1.51%)。NIR预测集的HPLC结果与NIR预测值之间未达到显著水平(p>0.05)。5、黄连中的生物碱成分与气候及土壤等生态因子之间存在较好的相关性。与各生物碱相关的主要气候因子是日照时数、极端最高温、极端最低温,主要土壤因子是水解性氮、有机质、速效磷及海拔高度。结论：1、四川产地的气候及土壤与其他产地存在明显的差异性；不同产地黄连样品在化学成分上有明显的差异性,药根碱、非洲防己碱与巴马汀可能是导致四川与其他产地黄连化学成分差异的主要因素。综合生态因子与化学成分的研究,初步建立了体现黄连道地性的质量评价方法,为其他的道地药材的研究提供参考。2、首次建立的HPLC同时测定黄连中6种生物碱的方法,可以更充分的控制黄连的质量。NIR直接对黄连粉末进行分析,能快速、无损、有效测定出黄连中的生物碱含量,为黄连的质量控制研究提供了新的研究方法。更多还原

【Abstract】 The coptis is rhizome of Coptis chinensis Franch., Coptis deltoidea C.Y.Cheng et Hsiao, and Coptis teeta Wall, which was first recorded in Shen Nong’s Herbal.The Coptis chinensis Franch. is commonly used in clinical practice. The samples of different origins have different quality. Because of the ecological environment, the chemical composition of samples of different areas has difference. The samples of Sichuan are famous-region drugs, because of their unique genetic background, ecological environment, cultural background and theory of TCM.Objective:Study the chemical composition and ecological factors of coptis, and explore the relationship between them. Establish the method of quality evaluation of coptis, which can reflect its genuineness of TCM.Method:1. Collect samples of different producing areas, study ecological factors of producing areas. Collect soil samples and determination their pH, organic matter, available N, P, K. Analysis difference of ecological factors of different areas.2. Establish NIR fingerprint of samples of different producing areas, use the PC A method to study the difference of samples.3. Establish HPLC fingerprint of samples, and determine the contents of six major alkaloids jatrorrhizine, columbamine, epiberberine, coptisine, palmatine, and berberine. Use the MPLS method to establish NIR calibration equation of six alkaloids and total alkaloids. Conduct paired two-tailed T test to the HPLC and NIR data of prediction set.4. Analyze the correlation of chemical composition and ecological factors by mathematical statistics methods (Hierarchical Cluster Analysis, Grey Correlation Analysis, Canonical Correlation Analysis and so on).Result:1. Collected 134 medicine samples and 19 soil samples of different producing areas. Cluster Analysis results show that samples of Chongqing and Hubei gathered to one group, and samples of Sichuan gathered to one group.2. The PC A result of NIR fingerprint shows that wild samples and processed samples of Sichuan are different with the samples of other producing areas, samples of four years and seven are different with samples of five years.3. HPLC fingerprints had detected 27 samples, and 12 common peaks had been calibrated, similarity is greater than 0.99. Cluster Analysis results show that samples of Chongqing, Hubei and Hunan gathered to one group, samples of Emei and Hongya gathered to one group, samples of Pengzhou, Chongzhou and Qionglai gathered to one group or gathered with samples of Emei and Hongya.4. The method for the identification and quantitative determination of six alkaloids had been established. The average recovery was 101.57%(RSD=1.33%),102.50 %(RSD=1.35%),100.80%(RSD=1.91%),102.66%(RSD=1.23%),97.85%(RSD=1.25% ),99.01%(RSD=1.51%).The results of HPLC and NIR of prediction set don’t reach significant level(p>0.05).5. There is good correlation between chemical composition and ecological factors, dominant factor are sunshine hours, highest and minimum temperature, available N, organic matter, available P and altitude.Conclusion:1. Climate and soil of Sichuan are different with other producing areas; coptis of Sichuan are different with other producing areas, and the reason maybe jatrorrhizine, columbamine, palmatine.Established the method of quality evaluation reflecting its genuineness of coptis. It can provide reference for other authentic herbs.2. The method of quantitative determination of six alkaloids which established firstly can explore the nature of chemical composition. NIR can determination medicinal powder directly, and can determination the main chemical composition fastly, losslessly and effectively, which is consistent with the overall concept of TCM. It can provide new method for quality control of TCM.更多还原