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枸杞叶化学成分及油果标准提取物的制备工艺研究
王艳
学位类型博士
导师阿吉艾克拜尔·艾萨
2016-06-03
学位授予单位中国科学院大学
学位授予地点北京
学位专业有机化学
关键词枸杞叶 化学成分 枸杞油果 多糖 多酚 提取 纯化
摘要

枸杞属于茄科 (solanceae) 枸杞属 (Lycium) 植物,在全世界分布广泛,约80种,我国分布有7种和3变种,其中以宁夏枸杞 (Lycium barbarum L.) 分布最为广泛,在宁夏、内蒙古、甘肃、青海、陕西、河北、新疆等地均有分布。我国除了有丰富的野生资源外,还对枸杞进行了大规模的种植,其中宁夏、新疆、河北、内蒙古是我国主要的枸杞产区。 枸杞自古以来就是可药食两用的植物,在我国有悠久的民间应用历史,其果实、根、叶均可入药,《本草纲目》、《神农本草经》和《本草汇言》中均有记载,枸杞具补肾、润肺、助阳、生精、益气、补虚、明目等功效。现代药理学研究表明枸杞叶具有良好的生物活性,但是其活性成分的物质基础尚不清楚,此外,由于传统的观念以及研究的不够深入,人们对枸杞叶的药用价值没有充分认识,以枸杞叶为原料开发的产品寥寥无几,造成药用资源的极大浪费。因此,对枸杞叶的化学成分进行深入系统的研究十分必要。另外,新疆拥有丰富的枸杞资源,但是产品的开发创新缓慢,目前绝大多数相关的产品开发仅限于简单的食品加工,相关的提取工艺、分离纯化方法等还没有十分深入的研究。因此本论文以枸杞叶和枸杞果实加工过程中的残次品-枸杞油果为研究对象,开展了化学成分研究、生物活性筛选和提取工艺研究,旨在加强对枸杞的科研开发,提高产品的附加值,为枸杞的进一步开发和综合利用奠定基础,具体工作如下: 1. 对枸杞叶的化学成分进行了系统的研究,分别从枸杞叶的氯仿部位、乙酸乙酯部位、正丁醇部位分离并鉴定了24个化合物,分别为莨菪亭 (LYC-1), 4-羟基苯丙烯酸乙酯 (LYC-2), 十六烷酸 (LYC-3), 邻苯二甲酸二丁酯 (LYC-4), 去氢催吐萝芙木醇 (LYC-5), 黑麦草内酯 (LYC-6), 丁香脂素 (LYC-7), lyciumnan (LYC-8), 吲哚甲酸 (LYC-9), 对羟基苯甲酸 (LYC-10), 对羟基桂皮酸 (LYC-11), 阿魏酸 (LYC-12), 咖啡酸乙酯 (LYC-13), 芦丁 (LYC-14), lyciumionoside A (LYC-15), lyciumionoside B (LYC-16), 3,4,6-trihydroxymegastigman-5-ene3-O-β-D-glucopyranoside (LYC-17), 云杉苷 (LYC-18), 东莨菪苷 (LYC-19), acanthoside D (LYC-20), lariciresineol 4′-O-β- D-glucopyranoside (LYC-21), lariciresineol 4-O-β- D-glucopyranoside (LYC-22), lyciumnanoside (LYC-23), N-trans-feruloyl tyramine-4′-O-β-D-glucopyranoside (LYC-24)。其中LYC-8, LYC-15, LYC-16和LYC-23为新化合物, LYC-2, LYC-5, LYC-6, LYC-7, LYC-9, LYC-13, LYC-17, LYC-18, LYC-20, LYC-21, LYC-22, LYC-24首次从枸杞属中分离得到。 2. 对枸杞叶的各极性部位进行了抗肿瘤活性和抗氧化活性测定,实验结果显示枸杞叶的石油醚部位、氯仿部位、乙酸乙酯部位和正丁醇部位对MCF-7, HeLa, A549, PC-3细胞株均有不同程度的细胞毒性,乙酸乙酯部位对A549和MCF-7细胞株的细胞毒性较强,正丁醇部位对HeLa和PC-3细胞株有较强的细胞毒性;分离得到的15个单体化合物对HeLa, A549, PC-3细胞株均有不同程度的细胞毒性;枸杞叶的正丁醇部位具有较好的DPPH 自由基清除能力,分离得到的LYC-5, LYC-12, LYC-13, LYC-14表现出一定的抗氧化活性。3. 以枸杞油果为原料,研究了枸杞油果提取物的提取和纯化工艺,以多糖和多酚的提取率为指标确定了最佳提取纯化条件为:枸杞油果粉碎石油醚超声脱脂后,以50%乙醇,料液比1:30,提取温度60 ℃,回流提取2 h,提取3次,提取浸膏以水溶解并稀释至0.17 g生药/mL上HPD-300大孔吸附树脂,上样流速2 BV/h,上样体积8 BV,以3 BV的蒸馏水除杂,洗脱剂为 50%的乙醇,洗脱剂用量3 BV,纯化后多糖含量为10.32%,多酚含量达到32.50%。

其他摘要

Goji was classified as solanceae, Lycium, widely grown throughout the world. It comprises eighty speices, and seven of them have been found in the People’s Republic of China. Lycium barbarum L. is exclusively distributed in Ninxia, Neimenggu, Gansu, Qinghai, Shanxi, Hebei, Xinjiang in China. Moreover, it has been cultivated on a large scale, especially the Ninxia, Xinjiang, Hebei, Neimenggu province is the majar prouducing regions. Goji were widely used as medicine and food in China, which has been used in Chna for a long history. In Chinese medicinal monographs “shen nong ben cao jing”, “ben cao gang mu” and “ben cao hui yan”, Goji was recorded as “nourishing liver and kidney, enhancing eyesight, enriching blood, invigorating sexreducing rheumatism” and so on. Modern pharmacological studies show that the leaves of Goji has good biological activity. Howerver, the material basis of its active components is not clear. Morever, traditional concept and the research is not deep enough, so people are not fully aware of the medicinal value of leaves. The prouducts with leaves of Goji as raw materia were very few. It caused a great waste of medical resources. Therefore, it is necessary to carried out a systematic study on the chemical composition of leaves of Goji. In addition, Xinjiang has rich resources of Goji. However, the development of the product is slow. The great majority of Goji procucts were confined to simple food processing. Extraction process and high efficiency extraction method not yet study thoutful. In this dissertation,the leaves and the black friuts of L. barbarum has been studied. We carried out a systematic study on the chemical composion and biological activities of leaves of L. barbarum, the extraction and purification of the polysacharids and phenolic of black fruits of L. barbarum. The aim of this work is to strengthen the research and development of Goji and meanwhile improve the added value of products. All this work lay the foundation for further scientific development and comprehensive utilization of Goji. 1. The systematic study on the chemical composition of leaves of L. barbarum. twenty-four compounds were seperated and identificated from the CHCl3, EtOAC and n-BuOH fractions. Four were new compounds, lyciumnan (LYC-8), lyciumionoside A (LYC-15), lyciumionoside B (LYC-16), lyciumnanoside (LYC-23) together with twenty known compounds, scopoletin (LYC-1), ethyl 3-(4-hydroxyphenyl) acrylate (LYC-2), palmitic acid (LYC-3), dibutyl phthalate (LYC-4), dehydrovomifoliol (LYC-5), loliolide (LYC-6), syringaresinol (LYC-7), indole-3-carboxylic acid (LYC-9), p-hydroxybenzoic acid (LYC-10), P-coumaric acid (LYC-11), feruic acid (LYC-12), caffeic acid ethyl ester (LYC-13), rutin (LYC-14), 3, 4,6-trihydroxymegastigman-5-ene3-O-β-D-glucopyranoside (LYC-17), picein (LYC-18), scopolin (LYC-19), acanthoside D (LYC-20), lariciresineol 4′-O-β- D-glucopyranoside (LYC-21),lariciresineol 4-O-β- D-glucopyranoside (LYC-22),N-trans-feruloyl tyramine-4′-O-β-D-glucopyranoside (LYC-24) were isolated from the leaves of L. barbarum. Compounds LYC-2, LYC-5, LYC-6, LYC-7, LYC-9, LYC-13, LYC-17, LYC-18, LYC-20, LYC-21, LYC-22 and LYC-24 were isolated from the plant for the first time. 2. The anti tumor and antioxidant activity of the different fractions of leaves of L. barbarum were determined. The results shows that the petroleum ether, CHCl3, EtOAC and n-BuOH fractions of leaves from L. barbarum displayed cytotoxicity on MCF-7, HeLa, A549, PC-3 cell lines. The EtOAC fractions of leaves of L. barbarum displayed a good cytotoxicity on A549 and MCF-7 cell lines. The n-BuOH fractions of leaves of L. barbarum displayed a good cytotoxicity on HeLa and PC-3 cell lines. Fifteen compounds displayed a different cytotoxicity on HeLa, A549, PC-3 cell lines. The n-BuOH fractions showed good antioxidant activity. Compounds LYC-5, LYC-12, LYC-13 and LYC-14 displayed antioxidant activity. 3. Study on the process of extraction and purification of extract of black fruits from L. barbarum. The methord was established with the polysacharides and phenols content as an indicator. The optimal extraction and purification were extract 3 times by 30 BV of 50% ethanol in 60 ℃, three hours each time. Then evaporated to dryness and diluted to 0.17 g crud drug/ mL, subjected to the HPD-300 macroporous resin column. The flow rate was 2 BV/h, and solution treatment capacity was 8 BV. After washing with 3 BV water, 70% ethanol was used as eluent, and the eluent volume was 3 BV at the flow rate 2 BV/h. After purification, the polysacharides and phenols content increased to 10.32% and 32.50%.

文献类型学位论文
条目标识符http://ir.xjipc.cas.cn/handle/365002/4584
专题资源化学研究室
作者单位中国科学院新疆理化技术研究所
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王艳. 枸杞叶化学成分及油果标准提取物的制备工艺研究[D]. 北京. 中国科学院大学,2016.
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