【作者】 南换杰；

【导师】 武滨； 秦雪梅；

【作者基本信息】 山西大学 ， 生药学， 2009， 硕士

【摘要】 黄芪是一种常用中药,具有补气固表,利尿托毒,排脓,敛疮生肌等功效。黄芪药材分布广泛,主要产区有甘肃、陕西、山西、宁夏、内蒙等省,质量差异较大,临床上又有各种炮制品使用,建立完善、综合的质量评价体系,并以此系统评价不同的黄芪药材及炮制品,对于有效控制其质量,为临床用药,以及中成药生产选择适宜的原料提供研究基础,具有重要的实用价值。本论文第一部分针对黄芪甲苷含量测定方法和指纹图谱分析中存在的一些问题,进行了修订完善;第二部分用系统评价体系,对黄芪的不同的品种、不同产地、不同采收年限、生品与炮制品等进行比较评价,获得大量的数据资料,主要研究结果如下:采用正交实验筛选黄芪甲苷含量测定样品前处理方法,优化了提取、纯化条件,即药材甲醇40mL浸泡过夜,超声提取2次,45min/次,合并提取液,挥干溶剂,残渣加水微热溶解,通过D101大孔树脂柱,经水洗,1%氢氧化钠溶液洗,再水洗至中性,然后依次用30%、95%乙醇洗脱,收集95%乙醇洗脱液,蒸干,残渣加甲醇使溶解作为供试品溶液。本方法与药典法不同之处在于,用多次超声提取代替索氏提取;直接过D101大孔树脂柱代替正丁醇萃取、碱洗及过柱;洗脱条件不同。含量测定结果与现行药典方法无显著性差异,但分析时间缩短2/3,回收率提高。此法应用于黄芪药材及复方制剂中黄芪甲苷的含量测定,方法可行。收集不同产地的黄芪药材,测定黄芪甲苷、总皂苷、总黄酮、多糖及部分微量元素的含量,结果表明:蒙古黄芪道地产区山西浑源以及内蒙古药材各成分含量较高,与其道地性品质相一致;主要引种栽培区甘肃药材质量接近道地产区,可考虑在甘肃进行规范化种植;不同基源的黄芪药材,主要有效成分含量一般蒙古黄芪高于膜荚黄芪。从单成分到大类成分多指标对黄芪化学成分进行含量测定,可以有效全面地控制黄芪质量。建立了黄芪药材乙酸乙酯与甲醇提取物高效液相指纹图谱:以乙腈—0.1%H₃PO₄水溶液为流动相,采用不同的梯度洗脱;流速:1mL/min;检测波长为210nm:柱温25℃。其中,乙酸乙酯提取物得到16个共有峰,甲醇提取物15个共有峰;方法学考察表明,该法的稳定性、重现性均符合指纹图谱分析技术要求,用中药色谱指纹图谱相似度评价系统软件分析结果显示:10批黄芪药材相似度＞0.8,说明各产地药材有比较好的一致性。试验证明,本方法可操作性强,重现性好,可作为黄芪药材指纹图谱研究的参考。与已有文献方法比较:采用了不同极性的溶剂提取,信息量较大,分离度及色谱峰形较好。采用相似度系数法、均值聚类法、主成分分析多种软件对指纹图谱进行分析计算,相似度结果表明:山西不同来源的黄芪药材相似度较高,说明道地产区黄芪药材化学组成一致性较好,质量稳定。均值聚类分析和主成分分析结果显示:蒙古黄芪和膜荚黄芪差异明显,说明从化学成分和指纹图谱可以初步鉴别不同物种之间的差异;相同的蒙古黄芪种中,道地产区山西浑源、内蒙古、主要栽培区甘肃黄芪和黑龙江聚为一类,表明聚类、主成分分析结果一致。通过正交试验确定了蜜炙黄芪烘制的工艺条件,综合多指标成分、紫外光谱、高效液相指纹图谱对比评价了烘制和炒制品及其他炮制品的质量,进行了相关性分析,表明蜜炙黄芪烘制品质量与传统的炒制品质量相近,同时对各种炮制品的功效与物质关系进行了初步的探讨,从化学成分的角度验证了传统炮制品功效不同的科学道理。更多还原

【Abstract】 Astragali is a commonly used Chinese Medicine, having the function of getting rid of sputum, diuretic, expectorant abilities. It is widely distributed in Gansu, shanxi, Ningxia, Neimenggu provinces and so on. There are various processed products of Radix Astragali, which are using in clinical. It has practical values to develop a complete and compositive Quality Evaluation System for evaluate quality of Radix Astragalus form different sources, and it is the basis of proper clinical administration or chosen of raw material in industry. The first part of this thesis revise the determination of astragaloside IV and analysis of Chromatography Fingerprint; the second part contains abundant data in Quality Evaluation System by comparing different species, different places of cultivation, different ages and seasions of collection, different procession of herb, and so on. The major results as follows:A suitable previous-processing method of astragaloside IV was screened by orthogonal experiment, and conditions of extraction and purification was optimized. Radix Astragalus samples was marinated in 40mL methanol all night before ultrasonic extraction. The extraction was repeated twice for 45 min. The extracted solution was mixed and evaporated to dryness. The residue was dissolved with water and eluted by water, 1% sodium hydroxide, water to neutral, as well as 30%, 95% ethanol through D101 Macroporous resin. The 95% ethanol elution was collected and evaporated. The residue was dissolved with methanol as working solution. The differences between this experiment and pharmacopeia are: ultrasonic extracting repeatedly instead of refluxing; the butanol extracting, alkali eluting and columning was replaced by cross D101 Macroporous resin directly; eluting conditions. Comparing with pharmacopeia method, there was no significance difference in the result. But analysis time is shorten 2/3 and recoveries was increased. Hence, this method could be applied to determine of Radix Astragalus and its compound. The contents of astragaloside IV, total saponines, total flavorids and certain microelements were determined after collecting Radix Astragalus samples of different places. The results demonstrate that astragaloside IV as the marker components in pharmacopeia is scientificalness for positive correlation among the major constituents of Radix Astragalus samples. Each content was higher in Shanxi hunyuan real estate area, so the quality is cosistent with its producing area. The quality of Neimeng is similar to that of Heilongjiang traditional regions. And quality of Ganshu cultivation is close to that of real estate area, so normalized cultivation could be considered in GanSu . It is overall available to control the quality of Radix Astragalus by chemical compositions from single to multi-component assaying.The method of HPLC fingerprint was established for the quality control of Radix Astragalus. The samples were extracted by Ethyl acetate and Methyl acohol separately and determined by HPLC with non-linear gradient elution. The mobile phase consisted of acetonitrile and 0.05% phosphoric acid. The flow rate was 1.0 ml/min, the wavelength was 210 nm and the column temperature was 25°C. The Ethyl acetate extraction of Radix Astragalus obtained 16 common peaks and Methyl acohol extraction obtained 15 common peaks. The chromatograms were analysised by "similarity evaluation system for chromatographic fingerprint of TCM" software, published by GPC(version 2004A). The fingerprint of ten samples were obtained and similarity more then 0.8 which had a high coherence. This method has a good stability. The fingerprint can overall reflect the quality of Radix Astragalus. Cmpared to the references, this method has more information and different solvent.The similarity coefficient method, mean clustering method, principal component analysis were used for the analysis of fingerprints. The similarity results show that: similarity degree was high in different sources of Shanxi and the chemical composition was consistent with each other, the Radix Astragali has better quality. Means clustering analysis and principal component analysis results show that: Mongolia Astragalus and Astragalus membranaceus has obvious differences, the differece among different species can be identificated from the chemical composition and the initial fingerprint. The same species, Shanxi hunyuan real estate area, neimeng, Heilongjiang traditional areas, the main cultivation areas in Gansu astragaloside were clustered, indicate that the result of clustering was consistent with principal component analysis results.Technological condition of honey processing was determined by orthogonal experiment. The quality of the baked, fried and other processed products was evalued through the integrated multi-index components, UV, HPLC fingerprint. The result of correlation analysis show that honey processing technology can replace the traditional frying processing, the effectiveness and material relationship of a variety of processed products were studied and verified the scientific principle of the effectiveness of different processed products from the traditional point of view.更多还原