锡金微孔草化学成分与生理生化特性的研究

NWIPB OpenIR

	锡金微孔草化学成分与生理生化特性的研究
	皮立
学位类型	博士
导师	韩发 ; 胡凤祖
	2014-05
学位授予单位	中国科学院研究生院
学位授予地点	北京
学位专业	生物学
关键词	锡金微孔草化学成分液质联用脱落酸生理生化
摘要	本文对锡金微孔草[Microula sikkimensis]化学成分和生理生化特性进行了研究。应用GC-MS、UPLC-MS/MS、ICP-OES等分析技术，对锡金微孔草主要化学成分进行了分析。从微孔草全草石油醚和正丁醇萃取部分提取、分离和鉴定五种化合物；用扫描电子显微镜，比较了不同海拔采样地点微孔草叶片的亚显微结构。为微孔草的活性成分开发和抗逆生理研究提供科学依据。本研究主要得到以下结论： 1.检测微孔草蛋白质，粗脂肪，多糖、总糖、脂肪酸、氨基酸、矿质元素和总黄酮的含量。全草中多糖（6.29%）和蛋白质（9.48%）的含量较高；氨基酸含量最高的是天冬氨酸（0.66%），其次是丝氨酸（0.61%）、赖氨酸（0.51%）；17种元素的平均值依K>Ca>P>Mg>Fe>Al>Na>Ba>Sr>Mn>Zn>B>Ti>Cu>Mo> Ni>Cr分布；总黄酮的平均值是4.65%，具有较高的开发价值。 2.对微孔草全草脂溶性成分和花挥发油进行了分析。脂溶性成分主要由硬脂酸(24.40%)、亚油酸(19.65%)、α-亚麻酸(19.86%)、β-谷甾醇（2.08%）等38个化合物组成。花挥发油的主要成分为醇类、烯类和酮类，主要由β-石竹烯(6.3%)、氧化石竹烯 (18.8%)、兰桉醇(7.3%)等34个化合物组成。3. 应用UPLC-MS/MS技术，分析微孔草全草提取液，鉴定出3种黄酮类成分，即芦丁、金丝桃苷、牡荆素。 4. 采用分散液液微萃取-高效液相色谱法测定微孔草叶中的脱落酸含量，为微孔草内源植物激素脱落酸的研究建立了方便、高效、省时的检测方法。 5. 用柱色谱法、薄层色谱法、重结晶法等分离手段，从微孔草全草石油醚和正丁醇萃取部位中分离出5种化合物，应用波谱技术（核磁氢谱、核磁碳谱）和参考文献数据进行了结构鉴定，分别是正十八烷醇(Ⅰ)、β-谷甾醇（Ⅱ）、芦丁（Ⅲ）、金丝桃苷（Ⅵ）和牡荆素（Ⅴ）。 6. 微孔草亚显微结构和抗逆生理的初步研究。随着海拔的升高，上下表皮的气孔密度和气孔长度均增加；气孔的长宽比，上表皮随海拔升高减少，下表皮的变化不明显。
其他摘要	The chemical component and physiological-biochemical characteristic of Microula sikkimensis were studied in this paper. we mainly studied the quantitative analysis of chemical constituents in M, sikkimensis by GC-MS, UPLC-MS/MS, ICP-OES and so on. Five compounds were isolated from the petroleum ether and n-butyl alcohol soluble part of ethanol extract of the herbs M, sikkimensis . Three flavonoids were identificated and analyzed in the whole plant of M. sikkimensis by UPLC-MS/MS. The determination method of abscisic acid was studied by DLLME-HPLC. Submicroscopic structure of leaves were compered in collect areas of different altitude by scanning electron microscope. The results provided the scientific basis for active ingridiant development and study of stress-resistant physiological. The conclusion are as follows: 1. The contents of protein, crude fat, polysaccharide, total suger, fatty acid, amino acid, mineral element and total flavonoids were analyzed. Polysaccharide (6.29%) and protein (9.48%) had higher contents in the herbs M, sikkimensis. The most abundant amino acid in the herbs M, sikkimensis is aspartic acid (0.66%),then serine (0.61%) and lysine (0.51%).The contents of 17 minerals in the herbs M, sikkimensis were determined by ICP-OES, the results of 17 minerals in the herbs showed K>Ca>P>Mg>Fe>Al>Na>Ba>Sr>Mn>Zn>B>Ti>Cu>Mo>Ni>Cr by mean values. The mean value of total flavones in the herbs M, sikkimensis is 4.65%, with great value for development. 2. Components of the liposoluble constituents and essential oil of flowers were first analyzed by GC-MS. liposoluble constituents mainly includes : octadecanoic acid(24.40%),linoleic acid (19.65%), α-linolenic acid (19.86%), β-sitosteri（2.08%）, etc. The components of essential oils of flowers were mainly included alcohols, alkenes and ketones. Caryophyllene oxide (18.8%),β-caryophyllene (6.3%), globulol (7.3%), α-eudesmol(7.9%) ，linalool (6.3%)，linalooloxide（3.2%）were major compounds found in the essential oils. 3. Three flavonoids extracted from M. sikkimensis had been identified by UPLC-MS/MS.The three flavonoids were rutin, hyperin and vitexin. 4. The minute amounts of abscisic acid of M. sikkimensis was determinated by DLLME-HPLC , built a kind test method of convenient and timesaving for the study of abscisic acid . 5. Five compounds were isolated from the petroleum ether(60-90℃) and ethyl acetate soluble part of ethanol extract by means of column chromatography(CC), thin layer chromatography(TLC), recrystallization, etc, and were identified as octadecyl alcohol (Ⅰ)、β-sitosterol（Ⅱ）、rutin（Ⅲ）、hyperin（Ⅵ）and vitexin（Ⅴ）by modern spectroscopic methods (1H-NMR, 13C-NMR), along with comparison to authentic samples or the data from literatures. 6. Preliminary study of submicroscopic structure and stress-resistant physiological was carried out on M. sikkimensis . With the increase of altitude, stomatal frequency and stomatal length were increased by different degrees ; With the increase of altitude, stomatal length-width ratio of epicuticula was decressed by different degrees, the change of stomatal length-width ratio of lower epidermis was not obvious.
文献类型	学位论文
条目标识符	http://210.75.249.4/handle/363003/4036
专题	中国科学院西北高原生物研究所
推荐引用方式 GB/T 7714	皮立. 锡金微孔草化学成分与生理生化特性的研究[D]. 北京. 中国科学院研究生院,2014.

条目包含的文件
文件名称/大小	文献类型	版本类型	开放类型	使用许可
锡金微孔草化学成分与生理生化特性的研究-（2453KB）			开放获取	CC BY-NC-SA	请求全文