Abstract Cordyceps, one of the well-known traditional Chinese medicines, is a composite consisting of the stromata of the fungus, Cordyceps sinensis (Berk.) Sacc. (Family: Hypocreaceae) parasitized on the larva of some species of insects (Family: Hepialidae). The polysaccharides are considered to possess antioxidant, immuno-modulation, anti-tumor, and hypoglycemic activities. However, most reports usually focused on the chemical and biological research on polysaccharides from cultured Cordyceps. To date, there is few study on the polysaccharide from natural Cordyceps sinensis, and the difference of polysaccharides between natural and cultured Cordyceps sinensis is still unknown. In addition, how to control the qualitiy of polysaccharides is a problem due to their complexity and similarity. In this study, a high-performance thin-layer chromatography (HPTLC) method was firstly developed to distinguish the polysaccharides from 6 traditional Chinese medicines (TCMs), including Cordyceps sinensis, Panax ginseng, Panax notoginseng, Panax quinquefolii, Ganoderma lucidum and Astragalus memberanaceus, using 7 monosaccharides and 2 glycuronic acids as references. Then, alkaline polysaccharides from natural Cordyceps sinensis were prepared and purified. The partial characteristics of purified polysaccharide were analyzed and elucidated. Finally, the antioxidant activities of crude alkaline extracted polysaccharides (CAP) and the purified polysaccharides (PCAP) were determined using DPPH and ABTS assay. There are three chapters in the thesis. Chapter 1 was a mini-review on Cordyceps polysaccharides related to the chemical research, biological activities, and qualitiy control. Chapter 2 described a high-performance thin-layer chromatography (HPTLC) method for the discrimination of polysaccharides from six traditional Chinese medicines. The acid hydrolyzates of polysaccharides were analyzed by HPTLC combined with two coloration methods and thin layer scanning technique. In this method, 7 monosaccharides and 2 glycuronic acids were used as reference compounds, and 6 crude polysaccharides were easily discriminated. The separation, purification, characteristics elucidation and antioxidant activity of CAP and PCAP were shown in Chapter 3. The techniques such as ultrafiltration, column chromatography were used, and a prue polysaccharide (PCAP) was obtained. The characteristics of PCAP, total sugar content, IR spectrum, molecular weight, constituted monosaccharide and their molar ratio, were analyzed using HPTLC, HPLC, and IR. DPPH and ABTS free radical scavenging assay was applied for evaluation the antioxidant activity of CAP and PCAP. Key words: Cordyceps sinensis, Alkaline extracted polysaccharides, Chromatography, Qualitiy control, Antioxidation IV

澳門大學碩士論文 摘要 冬蟲夏草 Cordyceps sinensis(Berk.) Sacc 是一種傳統名貴藥材,其性平, 味甘,入肺、腎經,具補肺益腎,止血化痰之功效。蟲草中的多糖類成分具有廣 泛的生物活性,如抗氧化,調節免疫,抗腫瘤,降血糖等。然而,近年來的研究 主要集中於對人工蟲草多糖的化學結構解析和生物活性測定。天然和人工蟲草多 糖是否一致,二者之間存在什麼樣的差異,如何對蟲草多糖進行品質控制成為擺 在我們面前的課題。 本課題首先建立了高效薄層色譜法分离7种單糖與2种糖醛酸旨在以冬蟲 夏草、人參、三七、西洋參、靈芝與黃芪6種中藥多糖為例探討中藥多糖鑒別方 法:随后對城提取天然冬蟲夏草多糖進行分離純化,並對得到一個城提多糖組分 進行了特性表徵分析;最後用DPPH 與ABTS 自由基清除實驗評價了天然蟲草 城提粗多糖和純化多糖的抗氧化活性。 論文共分為三個部分: 第一章為冬蟲夏草多糖文獻綜述,包括冬蟲夏草多糖的化學、生物活性、以 及品質控制的研究進展。 第二章為高效薄層色譜法鑒別6種中藥多糖, 實驗以7種單糖與2種糖醛酸 為參照,对多糖的酸水解产物进行分析,應用2种不同显色剂結合薄層掃描技術 建立了各多糖樣品的特徵譜用于定性鑒別。 第三章為天然蟲草城提多糖的分離、純化、特性分析与抗氧化活性研究。實 驗採用超濾、凝膠柱層析等方法獲得了一個純化的蟲草城提多糖,並應用高效薄 層色譜法(HPTLC)、高效液相色譜法(HPLC)、紅外光譜(IR)等方法對其, 特性包括:總糖含量、紅外特徵光譜、分子量、單糖組成及比例進行表徵分析, 最后采用 DPPH 與ABTS 自由基清除法考察了粗多糖與純化後多糖的抗氧化活 性。 關鍵詞 天然冬蟲夏草、城提多糖、色譜、品質控制、抗氧化 11