【作者】 王晓燕；

【导师】 韩正敏；

【作者基本信息】 南京林业大学 ， 森林保护学， 2010， 博士

【摘要】 石蒜属植物在我国分布的种类最多,约为该属的四分之一。石蒜属植物除具有较高的观赏价值外,其所含丰富的生物碱也得到广泛应用,尤其是加兰他敏生物碱,由于近年来临床用于老年痴呆病症早、中期的治疗而成为研究热点。本研究主要以忽地笑为试验材料,分别采用索氏提取、稀酸溶液提取、超声波辅助提取、微波辅助提取和超临界提取法对忽地笑中的加兰他敏生物碱进行提取,并对各提取方法进行了优化;比较了不同优化提取方法提取忽地笑中对加兰他敏提取效果;比较不同种源地忽地笑中加兰他敏含量的差异;分析了不同品种石蒜植物中加兰他敏的含量;对忽地笑材料进行了解剖观察;对忽地笑营养器官中的加兰他敏进行了组织化学定、荧光组织定和激光共聚焦技术的定位观察;对忽地笑进行了组织培养研究;对组织培养材料小鳞茎、愈伤组织和再生根进行了切片观察,对其加兰他敏生物碱进行了激光共聚焦定位研究。。主要研究结果如下:1、比较了各优化提取法提取忽地笑中加兰他敏的得率,以索氏提取最高,其次是超声波和微波辅助提取,酸性水溶液提取率最低;分析了同一植株的根、茎和叶等器官中加兰他敏的含量,其含量是根>鳞茎>叶;比较了不同地理种源地忽地笑中加兰他敏含量的差异,结果显示:湖北产忽地笑加兰他敏含量最高,安徽金寨忽地笑含量最低;对石蒜属植物不同物种加兰他敏的含量进行了分析比较,结果显示:中国石蒜中加兰他敏含量最高,其次是江西石蒜,忽地笑中含量最低。2、对忽地笑辅助提取液中加兰他敏等成分进行了GC-MS分析,经Xcalibur化学工作站,Nist2000标准质谱图库及人工图谱检索,共鉴定出8个化合物;对不同地理种源地的石蒜属植物的HPLC图谱数据进行聚类分析比较。3、对忽地笑营养器官切片进行了显微观察;以钼酸钠的浓硫酸溶液为显色剂对忽地笑的鳞叶切片进行了组织化学定位观察;选取340–360nm的范围波长为激发光,发射波长范围确定在430–500nm,对忽地笑的营养器官中的加兰他敏荧光显微观察;激光共聚焦显微技术对忽地笑营养器官中的加兰他敏进行了定位研究。4、对石蒜属植物的离体培养进行了初步尝试,并运用激光共聚焦显微技术对离体培养的小鳞茎、愈伤组织中的加兰他敏进行了定性分析,检测证明离体培养技术用于生产加兰他敏提取原料的可行性。本文通过对石蒜属植物中加兰他敏进行提取、定位和离体培养物提取加兰他敏可行性进行了研究,取得的结果可以为石蒜属植物资源利用等方面的研究提供基础和参考。更多还原

【Abstract】 About one quarter of Lycoris species are mainly distributed in China. In addition to a high ornamental value, it contains a wealth of alkaloids which has been widely applied. Recently, galanthamine alkaloids extracted from Lycoris has been used in clinical treatment of the early and medium-term Alzheimer’s disease.In this paper, lycoris aurea is taken as experimental material to extract galanthamine by using methods including Soxhlet extraction, diluted acid solution, ultrasonic-assisted extraction, microwave-assisted extraction, and supercritical fluid extraction, with each method being optimized. The paper also makes a comparison of effects of various optimized extraction methods and the difference of galanthamine content in the aurea from different origins. The paper analyzes the galanthamine content in the varieties of lycoris plants. The dissection of the aurea is carried out for further observation, and histochemistry location is conducted to observe the galanthamine contained in the scale leaf of the aurea. In this paper, fluorescence microscope technology is adopted to observe the sliced roots, scale leaf and lamina of the aurea, and Laser Scanning Confocal Microscope (LSCM) is adopted to observe the sliced bulblet, callus and regenerated roots obtained using tissue culture, in addition to the location of galanthamine for further observation.The main results are as follows:The extraction methods were compared: the Optimal method was Soxhlet extraction, the next methods was Ultrasonic-Assisted extractuion and Microwave-Assisted extraction, The least effective method was the Diluted acid.The galanthamine content in the root, stem and leaf were analysed, and the results showed the content in the root > stem >leaf. The galanthamine content which from different geographical provenances were compared: the hightest in aurea was from Hubei, the lowest was from Anhui Jinzhai. The content of different varieties were compared too: the hightest was L.chinensis Traub, the next was the L.radiata(L’her)Herb, the lowest was aruea.2. The composition of aurea extraction was analysed by GC-MS. 8 Compounds were Identified by Xcalibur chemical workstation, Nist standard MS Gallery and helped with artificial search, the data obtained by HPLC for different provenaces were analyzed.3. Vegetative organ of aurea were observed of by microscopy; the Slice of aurea were observed by Frode regent, the distribution of galanthamine in the tissue of Golden lycoris were localized: 340-360nm as the excitation,430-500 as the emission light, the distribution of galanthamine were observed Using fluorescence microscopy. The location fo galanthamine were studied by LSCM.4. An attempt to culture in vitro of Golden lycoris was conducted. Using of confocal laser technology, the content of galantamine was qualitative. The result proved that vitro technology for the production of raw materials for extracting galantamine was feasible.This paper addresses the feasibility of extraction and location of galanthamine in lycoris plants as well as the extraction of galanthamine from isolated culture products, and the results may provide basis and reference for the research on the use of lycoris plants resources.更多还原