【作者】 李洋；

【导师】 谭睿；

【作者基本信息】 西南交通大学 ， 微生物与生化药学， 2010， 硕士

【摘要】 大黄为临床常用中药,已有两千多年治病史,历代中药本草典籍中均有记载。在我国有45个品种和2个变种,主要分布于甘肃、青海、四川、陕西、西藏等地。2005版《中国药典》收录的大黄为蓼科植物掌叶大黄Rheum palmatum L.、唐古特大黄Rheum tanguticum Maxim.ex Balf.或药用大黄Rheum officinale Baill.的干燥根及根茎。大黄性味苦寒,泻热通肠、凉血解毒、逐瘀通经,传统用于抗菌、泻下,此外还具有广谱抗菌、保肝利胆、抗肿瘤、降血脂等广泛的药理作用,具有深厚的药用基础及巨大的开发潜力。综合现阶段对大黄药材质量控制研究文献,发现对于大黄药材质量控制方法较为集中于薄层色谱定性分析及高效液相色谱定量分析,但均是对于大黄药材中某一类或某一种药理活性成分进行检测。而大黄的药理作用发挥为多靶点,各成分相互作用而成。因此,上述方法未能体现出全面综合控制大黄药材质量,显示出一定的局限性。因此,本文从定量测定和定性分析角度切入,采用高效液相色谱法对29批不同来源的大黄药材分别进行3类不同药理活性物质中5个活性成分同时测定以及指纹图谱分析,获得较好结果。首先,本文通过HPLC条件优化筛选,建立了同时测定大黄药材中没食子酸、番泻苷A、芦荟大黄素、大黄素、大黄酚的HPLC含量测定方法,该方法稳定可靠,重现性良好。其中番泻苷A为主要泻下成分,其次为蒽醌类化合物；大黄素、芦荟大黄素抗菌作用最强；没食子酸、大黄酚主要发挥止血作用,对此5种化合物定量检测可基本实现从药理活性角度对大黄药材质量综合控制。实验中通过对药材供试品制备方法的优化筛选,本文确定了以甲醇为溶剂进行超声提取,无过多操作,快捷、准确、无损的含量测定供试品溶液制备方法,改善了现行供试品制备方法步骤繁多、耗时长、实际操作中易造成损失的弊端。其次,本文在对大黄HPLC色谱条件优化筛选后,初步建立了大黄HPLC指纹图谱。该方法可使样品中不同极性组分得到合适的保留,在75min内各组分全部出峰,色谱峰数达到50个左右。对29批不同来源大黄药材测定分析后,共检识出30个共有指纹峰,将大黄药材图谱划分为4个指纹区。实验结果进行比较分析后得知,各品种所含化学成分种类及含量均有一定差异,验证了大黄药材为多品种来源,故应针对不同品种建立相应的指纹图谱。本论文着重从HPLC含量测定和化学指纹图谱建立过程中色谱条件优化的方法和步骤进行大黄质量控制方法的探讨,力求从药理作用角度获得对大黄药材化学成分的定量含量和定性种类进行进一步研究,为建立全面、综合的大黄药材质量评价体系奠定一定基础。更多还原

【Abstract】 Rhubarb is a commonly used clinical medicine. It has a history of the millennium for treatment. In China there are 45 varieties and 2 varieties, it’s mainly distributed in Gansu, Qinghai, Sichuan, Shaanxi and Tibet. It is the dried rhizome of Rheum palmatum L., Rheum tanguticum Maxim.ex Balf. and Rheum officinale Baill. according to Pharmacopoeia of the People’s Republic of China (2005 edition).Rhubarb is traditionally used for purging heat, cooling blood, detoxification, break blood and expel stasis. In addition, Rhubarb also can be used to broad-spectrum antisepticize, protective and choleretic, anti-tumor, drop blood fat, etc. It has a wide range of pharmacological effects with strong foundation and tremendous medicinal potential.The recent comprehensive quality control of rhubarb is more focused on only one ingredient or one kind of ingredients by TLC and HPLC. However, the pharmacological effect of rhubarb is played by a class or a certain kind of pharmacological activity components. The above methods have showed some limitations in comprehensive quality control of rhubarb. Thus, a valid, simple and comprehensive quality control method is need to be established.29 batches of Rhubarb were determined with 5 pharmacologically active ingredients and done the fingerprint analysis in the basis of both quantitative and qualitative analysis by HPLC. The good results were obtained.Firstly, HPLC method was established for quantitatively simultaneous determining Gallic acid, Sennosides A, Aloe emodin, Emodin and Chrysophanol in Rhubarb. The method is stable and reliable, good reproducibility. In which Sennoside A is the strongest purgative component, followed by anthraquinone; Emodin and Aloe emodin has the strongest antibacterial activity; Gallic acid and Chrysophanol play the major role of hemostasis. So the simultaneous determination of the 5 kinds of basic compounds can consummated the Quality Control of Rhubarb from the view of pharmacological. Through testing and screening, the preparation method of medicinal products was selected. Only methanol extracted, without too much action. It was fast, accurate and nondestructive.Secondly, the HPLC fingerprint of Rhubarb was established. Different polarity ingredients were separated and retained the right time by the method. The run time is 75min and about 50 peaks were obtained.29 batches of Rhubarb were determined,30 distinguished peaks were found. The fingerprint was divided into 4 zones in order to do the identification of Rhubarb. After comparing the fingerprints, there were some differences between the 29 batches of Rhubarb. Because Rhubarb is from multi-source, fingerprint should be established for different varieties.This paper was focused on quality control of Rhubarb, through the research on chemical determination and HPLC fingerprint by choosing quality control conditions and steps. We try to do more research on quantitative chemical composition and qualitative content types of Rhubarb from the pharmacological point of view and lay a foundation for the establishment of a comprehensive, integrated system of quality control of Rhubarb.更多还原