Study On Extraction, Separation, Purification And Activity Of Flavonoids From Epimedium Koreanum

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 koreanumThe 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 HSCCCThe 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 ATo 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.