【作者】 关红晖；

【导师】 高英；

【作者基本信息】 广州中医药大学 ， 中药学， 2008， 硕士

【摘要】 “药对”,又称对药、对子,专指临床上相对固定的两味中药组成的方剂,是中药复方配伍中最简单、最基本和最常见的用药形式。药对配伍遵循中医“七情合和”理论组合法度,具备了复方的基本功能主治,药对是中药配伍方法的精髓,亦是药物组合方剂的核心,药对配伍的研究是复方配伍规律研究的基础和重要切入点之一:既具有复方的特性,又具有单味中药成分相对简单便于展开现代科学研究等特点。由此可见,研究中药复方的配伍规律可从药对着手。知母和黄柏是临床常用的清(虚)热药对之一。二者伍用最早出自会·李杲《兰室秘藏》滋肾丸,现代制剂中,有很多口服制剂中应用了知母黄柏的药对配伍,如知柏地黄丸等。鉴于药对对研究中药复方的配伍规律的重要性和知母黄柏药对配伍应用的普遍性,本课题选用知母黄柏药对作为工具药对进行研究,以期为进一步揭示应用知母黄柏的药对规律的现代制剂的物质基础打下良好的工作基础。本论文在综述了知母、黄柏及其药对国内外研究现状的基础上,主要研究内容包括:知母黄柏药材的质量研究;知母黄柏药对提取纯化工艺的研究;知母黄柏药对配伍前后化学成分的含量变化研究;知母黄柏的指纹图谱研究。1.知母黄柏药材的质量研究除进行药材显微鉴别和溥层鉴别、水分、总灰分检查外,还采用紫外可见比色法和高效液相法,以菝葜皂苷元、知母皂苷AⅢ和盐酸小檗碱为对照,测定药材知母中菝葜皂菅元和总皂苷、黄柏中盐酸小檗碱和总生物碱的含量。2.知母黄柏药对提取纯化工艺的研究提取工艺研究采用正交试验设计方法,考察了不同乙醇浓度,溶媒倍量,提取时间和提取次数对知母总皂苷和黄柏总生物碱提取率和纯度的影响,建立了四因素三水平的正交试验,最终确立了最佳的提取工艺为:8倍量70%乙醇加热回流提取3次,每次1小时。纯化工艺研究采用大孔树脂对知母总皂苷和黄柏总生物碱进行提取精制纯化,筛选了不同型号的树脂,考察了树脂的静态、动态吸附性能,并对上样液浓度和流速,洗脱溶剂浓度和体积等相关因素进行了优化,确立了最佳的纯化工艺为:XDA-5型大孔吸附树脂分离纯化知母总皂苷的最佳工艺为:样品液中总皂苷与干树脂重量比1∶8,依次用7BV蒸馏水、7BV80%乙醇以2ml/min的洗脱速度洗脱,收集80%L醇部分,减压浓缩至干,得到的总皂苷纯度在50.0%以上;确定Lx—18型大孔吸附树脂分离纯化黄柏总生物碱的最佳工艺为:样品液中总生物碱与干树脂重量比1∶15,依次用7BV蒸馏水、7BV50%乙醇以2ml/min的沈脱速度洗脱,收集50%乙醇部分,减压浓缩至干,得到的总生物碱纯度在50.0%以上。确定LX—18型大孔吸附树脂分离纯化药对总皂苷和总生物碱的工艺,得到的总皂苷总生物碱部位和的纯度在50.0%以上。3.知母黄柏药对配伍前后化学成分的含量变化研究采用高效液相法和紫外可见比色法,以菝葜皂苷元和盐酸小檗碱、知母皂苷AⅢ为对照,对知母黄柏药对配伍前后菝葜皂苷元和盐酸小檗碱、总皂苷和总生物碱的含量变化进行测定,以评价配伍对化学成分含量的影响。4.知母黄柏的指纹图谱研究对10个收集地的知母黄柏药材和相应的总皂苷和总生物碱纯化物进行了HPLC指纹图谱研究,通过色谱条件的优化建立了各自的指纹图谱及检测方法,以控制药材的质量。参照国家药监局颁布的《中药注射剂指纹图谱研究的技术要求(暂行)》及运用国家药典委员会中药色谱指纹图谱相似度评价系统(2004年A版)对指纹图谱进行了评价,结果显示,10批药材和相应的总皂苷和总生物碱纯化物的相似度都在0.9～1.0之间。还对药对进行指纹图谱研究,比较知母、黄柏配伍前后的化学成分。更多还原

【Abstract】 The drug couple,which is the most foundational and simplest form in the prescription compatibility of traditional Chinese medicine,specially imply the prescription composed by two relative fixed Chinese medicine in clinic.It posseses the basic indication function of the complex prescription.The drug couple is the marrow of compatibility,and also the core of the prescription composed with drug.The study of drug couple is the foundation and the cut-point in the investigation of the prescription compatibility.Because it not just has the characteristics of complex prescription,but also the feature of simple to spread out modern research.So we can investigate the regularity of compatibility begin with the drug couple.Rhizoma Anemarrhenae and Cortex Phellodendri are frequntly used in the compatibility of Traditional Chinese medicine.The earliest appearance of them is in the Zishen Pellet from《Lan shi mi cang》written by Li Gao in Jing-dynasty.The drug couple composed by the Rhizoma Anemarrhenae and Cortex Phellodendri were used in lots of modern preparation.Because of the significance in studying the regularity of compatibility and the catholicity of the application with Rhizoma Anemarrhenae and Cortex Phellodendri,we choose the drug couple composed of Rhizoma Anemarrhenae and Cortex Phellodendri as the object to investigate,to establish foundation of the study on the modern preparation.This article on the foundation of concluding study progess on Rhizoma Anemarrhenae Cortex Phellodendri and drug couple,mainly deals with quality studies on Rhizoma Anemarrhenae and Cortex Phellodendri,studies on the extraction and purification process,studies on the chemical composition before and after compatibility,fingerprint.1.Quality studies:besides micro-identification and TLC qualitative test,moisture,total ash test,using UV-colorimetry and HPLC,using berberine hydrochloride,timosaponin AⅢand sarsasapogenin as reference components, determin the contents of berberine hvdrochloride,total saponins,total alkaloids and sarsasapogenin.2.Studies were carried on the extractionand purification process of total saponins and total alkaloids by reviewing the impact of ethanol concentration, ratio of herb to solvent,extraction time and times.Set up the L₉（3¹）orthogonal test and confirmed the best extraction process as 8BV 70%ethanol distilling three times,1h per time.Extract of total saponins and total alkaloids was purified with macroeticular resins.Different kinds of resins were applied to review the static and dynamic adsorption ability,to optimize the concentration,volume and flow rate of eluant.It was confirmed that the best purification process was XDA-5 macroeticular resins,total saponins of eluant to dried resins being 1:8,eluted bv 7BY 80%ethanol after 7BV water with flow rate of 2ml/min.80%ethanol part was collected and dried under vacuum resulting in the total saponins content above 50.0%.It was confirmed that the best purification process was LX-18 macroeticular resins,total alkaloids of eluant to dried resins being 1:15,eluted by 7BV 50%ethanol after 7BV water with flow rate of 2ml/min.50%ethanol part was collected and dried under vacuum resulting in the total saponins content above 50.0%.It was confirmed that the best purification process was LX-18 macroeticular resins, total saponins and total alkaloids content above 50.0%.3.Studies on the chemical composition before and after compatibility: using UV-colorimetry and HPLC,using berberine hydrochloride and sarsasapogenin,timosaponin AⅢas reference components,to evaluate the influence to the component by the compatibility of the drug couple.4.Studies of HPLC fingerprint was conducted to ten origins of Rhizoma Anemarrhenae and Cortex Phellodendri,total saponins and total alkaloids purification.Their fingerprints were built up by optimizing chromatogram condition,to characterize common peaks separately,so that the quality of medicinal materials can be controlled.Referring to "Technical Specifications of Fingerprint Study of Chinese Traditional Medicine Injection （Interim Regulation）" decreed by SFDA and similarity evaluation system for chromatographic fingerprint of TCM（2004 A）set by China pharmacopoeia committee,the results showed the similarity of Rhizoma Anemarrhenae and Cortex Phellodendri,total saponins and total alkaloids purification were both from0.9～1.0.To evaluate the influence to the component by the compatibility of the drug couple by using drug couple fingerprint.更多还原