【作者】 伍蔚萍；

【导师】 孙文基； 阎宏涛；

【作者基本信息】 西北大学 ， 分析化学， 2004， 硕士

【摘要】 甘草中的主要有效成分为皂苷类和黄酮类，而以往人们仅对甘草中皂苷类成分作了大量深入的研究，评价甘草的质量也仅以甘草酸的含量作为其质量优劣的标准，显然不够全面。同时，随着药理学的深入研究，表明甘草黄酮类成分是甘草的一类主要活性成分，具有抗炎、抗HIV、抗肿瘤等多种药理作用和广阔的开发及应用前景，已成为当前中药甘草研究的热点。本文结合05年版药典任务—对甘草中黄酮类成分进行研究及其含量测定，为建立甘草黄酮类成分的质量标准提供理论依据。 全文由以下五部分组成： （一） 综述 对甘草黄酮类成分的研究进展及中药质量标准研究状况进行了综述。 （二） 甘草黄酮类成分的提取与分离 将甘草药材粉末用水煮醇沉法提取，再依次将甘草水溶液用乙酸乙酯和正丁醇溶剂萃取，乙酸乙酯层提取液分别经硅胶柱和聚酰胺柱层析分离后，均可得到同一单体化合物。经化学方法和光谱解析鉴定为已知成分甘草苷，为甘草黄酮类的一个主要成分。结果表明，采用硅胶柱层析分离比聚酰胺柱层析较易获得甘草苷；采用样品系陕西合阳县所产，其甘草苷的含量较高。 （三） 2005年版中国药典甘草含量测定项标准修订 结合2005年版中国药典甘草含量测定项标准修订任务。选用超声法处理样品，应用高效液相色谱法，以乙睛-0.5％醋酸（1：4）为流动相，检测波长为276nm，对24份甘草样品中甘草苷含量进行了测定。结果表明甘草苷含量最高可达3.72％，最低为0.86％，其平均含量达1％以上，所以规定甘草中甘草苷的含量限度不低于1.0％。该方法线性关系良好（r=0.9998），最小检出限为0.1μg·mL^-1，回收率在97.66％～103.5％之间，该方法具有操作简便、快速，准确等优点，并就此拟定了2005年版药典甘草含量测定项正文（草案）。 （四） 紫外分光光度法测定甘草中总黄酮的含量 利用二氢黄酮类化合物在碱性条件下转化为查尔酮类的特性，以甘草苷为对照品，10％KOH溶液作为显色剂，在波长400nm处进行测定，建立了测定甘草中总黄酮含量的紫外分光光度法，并应用该方法对46份甘草样品中的总黄酮含量进行了测定。表明该方法的线性关系良好（件0.9992），回收率在95.0%一100.5%之间，46份甘草样品的总黄酮平均含量为4.37%。该法与通常测定方法相比，具有操作简便，准确度高，重现性好等优点。 （五）甘草黄酮类成分的指纹图谱研究采用HPLC方法，选用70%乙醇超声提取、乙酸乙酷萃取法处理样品，以乙睛一0.2%醋酸为流动相，梯度洗脱，检测波长为310nm。在选定的色谱条件下，考察了10批不同产地的甘草药材，标示出n个色谱峰作为甘草黄酮类成分指纹图谱的共有特征峰，并使用几种不同的相似度计算软件对结果进行评价。结果表明不同产地的甘草药材其化学成分存在差异，但各共有峰均存在，相似度在0.78卜0.931之间。通过建立甘草黄酮类成分的指纹图谱，可有效地鉴别甘草药材真伪，为全面评价及控制甘草内在质量提供新的方法和理论依据。更多还原

【Abstract】 The glycyrrhizin and flavonoids are one of the important components in Licorice, However, people had been put the high emphasis on glycyrrhizin in the past. So the content of glycyrrhizic acid was only determined in Licorice for appreciating the quality of Licorice. Obviously, It’s difficult to reflect the quality of Licorice. On the development of the study on Licorice, these pharmacological actions of the flavonoids in Licorice have been shown, such as anti-inflammation, anti-tumour, ant-virus and so on. It’s very important for studying and exploiting the flavonoids in Licorice. To fulfill the mission of the People’s Republic of China Pharmacopoeia of 2005, that is to say, the determination of the chemical of flavonoids in Licorice will be added. This thesis includes five parts as follows:Part I: Review. A review of the flavonoids in licorice and the quality standard of Chinese traditional medicine were given.Part II: Extraction and separation of the flavonoids from the rootstalk of Licorice. The medicinal material power was extracted by boiled water, and then the ethanol was added to water solution for depositing some impurities. The ethyl acetate and 1-butanol were added to extract the water solution respectively. The silica gel column and the polyamide column were used to separate the part of ethyl acetate in turn. At last, a same compound was gained from the two column respectively, which was liquiritin identified by chemical and spectral method. The result showed that liquiritin was easily gained by using the silica gel column and the content of liquiritin was high in Licorice from the He-yang County of Shaanxi Province.Part III: Recension the content of Licorice of the People’s Republic of China Pharmacopoeia of 2005. The sample was extracted by ultrasonic method, which the reagent was seventy-percent ethanol. A method for determination of liquiritin in Licorice by HPLC was established. The mobile phase was consisted of acetonitrile-0.5% acetic acid (1:4) and the detection wavelength was at 276nm. Then thecontent of liquiritin in twenty-four samples was determined by this method. The result showed the content of liquiritin was between 0.86%~3.72%, the linear correlation was good (r=0.9998) and the average recovery was between 97.66%~103.5%. This method was simple, reproducible and rapid, which was suitable for determination of liquiritin in Licorice. According these results, the text (draft) of the determination content item on Licorice for the People’s Republic of China Pharmacopoeia of 2005 was established.Part IV: Determination the content of total flavones in Licorice by UV-spectrophotometer. Based on the characteristic that flavanonols are easily transformed chalcones under alkali condition, a UV-spectrophotometry was developed. By this method, the content of total flavones was determined for forty-six samples. The result showed the average content of total flavones in Licorice was 4.37%, the linear correlation was good (r=0.9992) and the average recovery was between 95.0%~100.5%. This method was simple, reproducible, accurate comparing the usual method.Part V: Study on the fingerprint of the flavonoids in Licorice. The ultrasonic method was used to extract samples, which the reagent was seventy-percent ethanol. Then the ethyl acetate was added to extract the water solution of sample. The mobile phase was consisted of acetonitrile-0.2% acetic acid and detection wavelength was at 31 Onm. Under grads eluting condition, ten patches of samples from different producing area were determined. The result showed that the eleven mutual characteristic peaks were detected for ten patches of samples. At the same time, different similarity software systems were adopted to appraise the result. The result showed the similarity was between 0.781~0.931, which indicated these samples existed difference. By establishing the fingerprint on flavonoids of Licorice, the genuine or fake from the medicinal material was differentitated. In a word, a new method controlling the quality of Licorice was established.更多还原