【作者】 姚成；

【导师】 欧阳平凯；

【作者基本信息】 南京工业大学 ， 生物化工， 2003， 博士

【摘要】 对中草药有效成分的提取和分离方法、化学成分的主要分析方法和中草药特征指纹图谱的研究等方面进行了较详细的概述，对当归和猫爪草的化学成分分析和药理方面的国内外研究现状进行了详细综述。在此基础上，针对目前中草药化学成分分析研究均采用某一方法或技术,由于这些方法或技术的局限性，不能全面反映中草药复杂的化学成分，提出了采用综合分析方法进行中草药化学成分分析的新思路，以期对中草药的化学成分作全面分析研究。采用气相色谱-质谱联用技术（GC-MS）、电感偶合等离子体原子发射光谱（ICP-AES）、高效液相色谱（HPLC）、高效液相色谱-质谱联用技术（HPLC-MS）、氨基酸分析仪等现代分析手段、结合衍生化和分离技术的综合分析方法，对甘肃当归和不同产地的猫爪草进行了化学成分综合分析研究。首次采用GC-MS分析了河南产猫爪草乙醚提取物的主要化学成分，分析鉴定出：5-羟甲基二氢呋喃-2-酮、8-十八碳烯酸甲酯、油酸乙酯、亚油酸乙酯、亚麻酸乙酯、decahydro-1,5,5,8a-tetramethyl-1,2,4-methanoazulene、decahydro-4,8,8-trimethyl- 9- methylene-1,4-methanoazulene等物质；采用柱前衍生化方法并结合GC-MS，分析并鉴定猫爪草中含有：壬二酸、邻笨二甲酸、肉豆蔻酸、十五烷酸、棕榈酸、棕榈烯酸、十七酸、硬脂酸、8-十八碳烯酸、油酸、亚油酸、α-亚麻酸、二十烷酸、二十二烷酸、二十三烷酸等有机酸、胆甾醇、豆甾醇、豆烯甾醇、菜油甾醇等四种甾醇和葡萄糖、乳糖、麦芽糖等三种糖。其中多种成分为首次从猫爪草中被发现；首次采用ICP-AES方法分析了河南和新疆产猫爪草中的21种微量元素含量，同时进行了干法和湿法对比。结果表明：两种产地的猫爪草均含有人体必需的微量元素Fe、Co、Cu、Mg、Mn、Zn、Ni 、Se、Sr等，其中Zn高Cu低的现象与许多抗癌中药中Zn、Cu趋势一致，Se被公认为对肿瘤具有预防和抑制作用；首次采用氨基酸分析仪分析了河南、新疆、山东、湖北和安徽5个产地猫爪草中氨基酸的种类和含量，发现猫爪草中含有15种以上的氨基酸，不同产地猫爪草中所含氨基酸种类基本一致，含量与产地有关。采用加压柱层析方法从河南猫爪草乙醇提取液中分离得到了棕榈酸、β-谷甾醇、豆甾醇和β-谷甾醇-D-葡萄糖甙四种纯物质，并用红外光谱、熔点、薄<WP=4>层色谱、纯物质标准图谱等方法对其进行了鉴定。采用GC-MS分析了甘肃岷县当归挥发油的主要化学成分，分离得到72个色谱峰，鉴定出其中44个成分，其中32个成分为首次从岷县当归中发现；采用ICP-AES法测定了甘肃岷县和陇西当归中23种微量元素的含量，并对两种产地的微量元素测定结果进行了对比；对甘肃岷县和陇西当归及不同部位的氨基酸进行了分析，结果表明，陇西当归氨基酸含量大于岷县当归，陇西当归的不同部位氨基酸含量为：归头>归身>归尾；采用柱层析方法从甘肃岷县当归乙醇提取液中分离得到了十四醇-1、棕榈酸、阿魏酸、丁二酸等四种纯物质，同时对其进行了鉴定。对HPLC-MS联用技术在当归提取液成分分析方面的应用进行了探索性研究。对甘肃岷县和湖北兴山产当归乙醇提取液的HPLC分离的流动相条件进行了选择和优化，确定了三元梯度洗脱的流动相组成和比例；在此基础上采用电喷雾HPLC-ESI-MS对湖北兴山当归和甘肃岷县当归中的成分进行了分析，依据已知的五种有效成分的信息，对色谱和质谱图中相应的谱图提出了解释，为进一步进行该方法的深入研究奠定了基础。首次采用衍生化毛细管气相色谱法对不同产地猫爪草中的葡萄糖、乳糖、麦芽糖等糖及总糖和胆甾醇、豆甾醇、豆烯甾醇、菜油甾醇等四种甾醇的含量进行了分析，结果表明，不同产地的猫爪草中所含糖的种类、含量和总糖的含量不同；不同产地猫爪草中甾醇的含量也有不同。研究结果表明，采用综合分析方法进行中草药化学成分的分析研究，分析结果更全面，更能全面反映中草药中所含的化学成分。采用中草药化学成分的综合分析研究，对推动中草药化学成分的分析研究进展，促进中草药药理研究和新药开发具有重要的理论意义。更多还原

【Abstract】 Based on the summary review of the extraction and isolation of pharmaceutical components from the Chinese traditional medicinal plants, and the instrumental analysis of the chemical constituents they contained, and the research about the fingerprint of Chinese traditional medicine, the present situation of analysis of chemical constituents in Angelica Sinensis (Oliv.) Diels (Dnaggui in Chinese) and Ranunculus ternatus Thunb, and their pharmacological research were discussed in detail. Because of the limitation or disadvantage resulted from the single method or technique used in the analytical study of the components of Chinese medicinal plants, the complex constituents in the herbs can not be evaluated comprehensively. Therefore, in order to have a comprehensive and systematical knowledge and evaluation of the constituents in the Chinese medicinal plants, the author proposed a new idea of using an integrated analytical method to study and evaluate the constituents in the Chinese medicinal plants. Using the modern instrumental analytical methods such as GC-MS, ICP-AES, HPLC, HPLC-MS, Amino acid analyzer, combined with the derivation and separation techniques, a comparative and comprehensive analytical study of the Angelica Sinensis (Oliv.) Diels and Ranunculus ternatus Thunb from different production areas was performed. For the first time, the main chemical constituents in the ether extracts from Ranunculus ternatus Thunb grown in Henan Province were analyzed by GC-MS. Components were determined as following: 5-hydrmethyldihydrofuran-2-one, 8-Octdecenoic acid methyl ester, decahydro-1,5,5,8a-tetramethyl- 1,2,4- methanoazulene, decahydro-4,8,8- trimethyl-9-methylene-1,4-methanoazulene, ethyl oleate, linoleic acid ethyl ester, etc.. Then, the extracts of Ranunculus ternatus Thunb were derived and determined by GC-MS. The results indicated that it contains fatty acids such as: nonanedioic acid, phthalic acid, myristic acid, pentadecanoic acid,hexadecanoic acid, 9-hexadecenoic acid, heptadecanoic acid, octadecanoic acid, 8-octadecenoic acid, 9-octadecenoic acid, 9,12- octadecenoic acid, 9,12,15-octadecatrienoic acid, eicosanoic acid, docosanoic acid, tricosanoic acid, and four sterols: Cholesterol, Campesterol, Stigmasterol, Stigmastenol, and three<WP=6>monoglucosides : glucose, lactose, maltose. Many constituents were detected for the first time. With ICP-AES, 21 kinds of trace elements in Henan and Xinjiang Ranunculus ternatus Thunbs were respectively determined in both dry-ashing treatment method and wet one. Through comparison, it indicates that Ranunculus ternatus Thunbs from the 2 locations both contain trace elements which are necessary for health of human beings such as: Fe, Co, Cu, Mg, Mn, Zn, Ni, Se, Sr etc. The result that the content of Zn is higher than that of Cu is consistent with the relative content level of Zn and Cu contained in many anticancer Chinese traditional medicines. Moreover, as we all known, Se is helpful for the prevention and prohibition of tumors. The amino acids in Ranunculus ternatus Thunbs from Henan, Xinjiang, Shandong, Hubei, Anhui provinces were comparatively analyzed by the amino acid analyzer. It was found that: Ranunculus ternatus Thunb contains more than 15 kinds of amino acids; the sorts of amino acids in Ranunculus ternatus Thunbs from different locations are similar, but the contents of them are different from their locations. With the technique of flash chromatography, four compounds were isolated from the ethanol fraction of Ranunculus ternatus Thunb from Henan. Through the inspection of melting point, FT-IR, TLC, comparison with standard spectrum, they were identified as palmitic acid,β-sitosterol, stigmasterol andβ-sitosterol-D-glucoside. Using GC-MS, the volatile oil extracted from Angelica Sinensis of Minxian, Gansu province was analyzed directly. 44 compounds were identified from 72 isolated constituents, among which 32 compounds were found for the first time in Gansu Danggui. Meanwhile, 23 kinds of trace elements in Angelica Sinensis from M更多还原