Herba Epimedii, Yinyanghuo in Chinese, is one of the commonly used Chinese medicines for a long time. According to Chinese Pharmacopoeia, the dried aerial parts of Epimedium brevicornu Maxim., Epimedium sagittatum (Sieb. et Zucc.) Maxim., Epimedium pubescens Maxim., Epimedium wushanense T.S.Ying and Epimedium koreanum Nakai are used as Yinyanghuo, which has the effect of anti osteoporosis, immunological function modulation and anti-tumor actions, etc. In present study, fifteen flavonoids were separated and identified, and HPLC, UPLC and CZE methods were developed for the qualitative and quantitative analysis of flavonoids in Epimedium. The characteristics of different analytical methods were also compared. The thesis consists of seven chapters. Chapter 1 reviewed the advance in the research of flavonoids components and analytical methods of flavonoids in Epimedium. Chapter 2 focused on the separation of chemical compounds from Epimedium. Fifteen flavonoids were isolated and purified from total flavonoids extract of Epimedium by various chromatographic methods. They were identified as baohuoside I, 2"-O-rhamnosyl icariside II, epimedin C, icariin, hexandraside F, epimedin A, hexandraside E, epimedin B, caohuoside C, sagittatoside A, baohuoside VII, kaempferol-3-O-rhamnoside, sagittatoside B, baohuoside II and epimedoside C, which provided the reference substances for the analysis of Epimedium. In Chapter 3, The parameters of pressurized Liquid Extraction (PLE) including the type of solvent, extraction temperature, pressures, static extraction time, flush volume, cycles, times and particle size were optimized by using univariate approach. Then an HPLC-DAD-MS method was developed for qualitative and quantitative analysis of Epimedium. 26 samples were analyzed and the results showed that the contents of flavonoids were greatly variant in different species or locations of Epmedium. Simultaneous analysis of multiple components is more reasonable for quality control of Chinese medicine. Hierarchical clustering analysis based on characteristics of 15 investigated compounds peaks in HPLC profiles showed that 26 samples were divided into three main clusters, which in accordance with their flavonoids contents. Four flavonoids including epimedin A, B, C and icariin were optimized as markers for quality control of the species of Epimedium used as Yinyanghuo. Chapter 4 developed a UPC method for quality control of Epimedium. Fifteen flavonoids in 37 samples were determined using the validated method. Simulative mean chromatograms were generated based on the hierarchical clustering analysis. The correlation coefficients of each sample to mutual plot of high content cluster were compared. Generally the results were consistent with that of hierarchical clustering analysis. Chapter 5 is a primary study of CZE method for simultaneous determination of sixteen flavonoids in Epimedium and 26 samples were determined using the validated method. Chapter 6 compared the characteristic of HPLC, UPLC and CZE. The results showed that the three methods have their advantages and shortages respectively. When doing analysis, the most suitable method should be chosen depending on the requirement of analysis and condition of laboratory. Chapter 7 was a summary of this study. Key words: Epimedium; flavonoids; HPLC-DAD-MS; UPLC-PDA; CZE-DAD

淫羊藿 (Herba Epimedi） 是我國使用最為悠久的中藥之一，別名仙靈脾、三枝九葉草等，始載于《神農本草經》，列為中品。具有補腎陽，強筋骨，祛風濕的功效，用於陽痿遺精，筋骨萎軟，風濕痹痛，麻木拘攣以及更年期高血壓的治療。《中國藥典》2005年版一部收載的淫羊藿為小檗科植物淫羊藿Epimedium brevicormum Maxim.、箭葉淫羊藿Epimedium sagittatum (Sieb. et Zucc.) Maxim.、柔毛淫羊藿Epimedium pubescens Maxim.、巫山淫羊藿Epimedium wushanense T.S.Ying或朝鮮淫羊藿Bpimedium koreanum Nakai.的乾燥地上部分。為了對淫羊藿進行系統的品質分析，建立有效的品質控制方法，本課題在對淫羊藿進行化學成分研究、製備相應的對照品的基礎上，對其進行了系統的定性、定量分析，並對不同方法進行了比較。 全文共分七章。第一章對中藥淫羊藿的黃酮類化學成分和含量測定方法進行了詳細的綜述。第二章利用現代多種色譜分離技術，對淫羊藿總黃酮提取物進行了化學成分研究，分離鑒定了15個化合物，分別為：寶藿苷I (baohuoside I)、鼠李糖基淫羊藿次苷I (2”-O-tharnosyl icariside II)、朝藿定C (epimedin C)、淫羊藿苷 （icariin)、溫哥華苷F (hexandraside F)、朝藿定A (epimedin A)、溫哥華苷E(hexandraside E)、朝藿定B (epimedin B)、朝藿苷C (caohuoside C)、箭藿苷A(sagittatoside A）、寶藿苷 VII (baohuoside VII、山柰酚-3-O-鼠李糖苷(kaempferol-3-O-rhamnoside)、箭藿苷B (sagittatoside B)、寶藿苷II (baohuoside II)和淫羊藿苷C (epimedoside C)。為淫羊藿定性、定量分析方法的建立奠定了物質基礎。第三章首先對淫羊藿的加壓溶劑提取條件進行了優化。對提取溶劑的種類、提取溫度、壓力、時間、沖洗體積、週期、次數和樣品粒徑等進行了單因素考察，確定了淫羊藿藥材的加壓溶劑提取條件。之後採用HPIC-DAD-MS法建立了淫羊藿黃酮類成分的定性、定量分析方法，對全國各產地藥材進行了测定，並對結果進行了聚類分析，結果表明，26批藥材樣品根據黃酮含量的高低可分為三類，cpimedin A，B，C和icariin被優化作為淫羊藿品質控制的指標性成分。第四章採用UPLC-PDA法建立了淫羊藿中黄酮類成分的含量測定方法，對全國各產地37批藥材樣品進行了含量測定。在聚類分析的基礎上，建立了高含量組樣品的共有圖譜，並將各藥材圖譜與其比較，進行相似度評價，結果發現高、中、低含量組的樣品相關係數分別為0.92±0.05、0.70±0.15、0.18±0.03，與聚類分析的結果基本一致。第五章對CZE-DAD法分析淫羊藿中黃酮類成分的方法進行了初步研究，建立了CZE測定淫羊藿中黄酮類成分含量的方法，測定了26批樣品中的16個黃酮類成分。第六章對HPLC、UPLC和CZE分析淫羊藿中黃酮類成分的優缺點進行了比較，結果發現三種方法各有其特點，在應用時應結合具體分析的要求和實驗室的條件選擇最適合的分析方法。第七章是研究結果的總結與討論。關鍵詞: 淫羊藿；黄酮；HPLC-DAD-MS; UPLC-PDA；CZE-DAD