摘要
朝鲜淫羊藿是我国药典收录的传统中药,是生长在长白山地区的道地药材。大量基础研究和临床实践证明,朝鲜淫羊藿具有温补肾阳、强筋骨、祛风湿之功效。
在本论文中,首先建立了朝鲜淫羊藿总黄酮提取的简便且有效的方法,然后,将总黄酮在一定条件下进行转化,得到羊藿苷A粗提物。粗提物经硅胶柱纯化后,可以得到80%～85%的羊藿苷A。同时,建立了高速逆流色谱(HSCCC)分离羊藿苷A单体的方法,以此单体化合物为标准品对提取物中的羊藿苷A进行定性定量分析。最后,对羊藿苷A进行了初步的药效学研究,实验结果表明,羊藿苷A对实验性心肌缺血具有保护作用。
目前,人们对朝鲜淫羊藿中所含化合物的研究比较多,对其中所含化合物的种类与结构的研究比较深入。本论文提供了从朝鲜淫羊藿中提取含量高的羊藿苷A粉末,且进一步纯化的方法。纯化后的羊藿苷A组分简单,质量易控,药效好,为新药的研发打下了坚实的基础。
Epimedium koreanum is included in the traditional Chinese medicine in China Pharmacopoeia which is the growth in the Changbai Mountain area. Many basic researches and clinical practices shows that the Epimedium koreanum have Warming kidney, strengthening muscles and eliminating Rheumatism. In present thesis, the extraction of flavonoids in Epimedium koreanum, purification and activity of icariside were studied.
1. The extraction and purification of flavonoids in Epimedium koreanum
The Epimedium koreanum leaves were smashed and extracted twice with 70% ethanol solution. The content of total flavonoid in extract was determined by UV-Vis spectrophotometer. Icariin was used as the standard. The yield of total flavonoids in Epimedium koreanum is12%. Extract was heated. Epimedium flavonoids would become Yanghuogan A . 5 g of crude extracts were got from 100 g of leaves, of which Yanghuogan A content could be up to 20%～30%. Extracts were purified by silica gel column which were eluted by chloroform - methanol -wate(9:10:0.2, v/v/v). Yanghuogan A was detected by TLC detection. The expand agents is chloroform - methanol– water (9:10:0.2, v/v/v). Yanghuogan A elution was collected and determined by HPLC method. The results showed that the yield of Yanghuogan A could be up to 60%～70%. The content of Yanghuogan A in elution could be up to 80%～85%.
2. Purification of Yanghuogan A by HSCCC
The powder of Yanghuogan A can be isolated by high-speed countercurrent chromatography (HSCCC). Using hexane - methanol - water(5:4.5:5.5, v/v/v) as the two-phase solvent system. The upper was used as stationary phase and the lower as mobile phase. Flow rate is 2.0 ml/min. Temperature is 30℃. Retention rate in stationary phase is 50 %. We can get Yanghuogan A whose purity is 99 %. Qualitative and quantitative analysis of the extract were carried out by using Yanghuogan A as standard. Between 0.04～0.36mg/mL, Yanghuogan A has the good linearity, a better recovery and precision.
3. Primitive study on activity of Yanghuogan A
To explore the activity of Yanghuogan A, the protective pharmcology functions of Epimedium Flavonoids were compared to compound danshen dripping pills in the pituitrin-induced rat myocardial ischemia model. After disease induction, the 40 rats were randomly assigned to the four experimental groups: one group rats were orally administered with water as control group and the other groups were administered with total flavonoids, Yanghuogan A and compound danshen dripping pills respectively. Once a day at 200 mg/kg for seven days. We found that T wave of the flavonoids and Yanghuogan A groups was lower than that of control and the effect of Yanghuogan A was better than flavonoids. The result shows that Yanghuogan A has the functions of anti-acute myocardium ischemia.
引文
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