露兜簕果中有效成分的研究
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摘要
露兜簕(Pandanus tectorius Sol.)又名野菠萝,广布于亚洲、非洲和大洋洲热带和亚热带地区,其根、叶、花、果都可以药用,本论文通过对露兜簕果的有效成分及相关研究,更好地分析露兜簕潜在的资源价值,为开发利用这一植物资源和科学研究提供一些依据,主要研究内容与结果如下：
1.取新鲜露兜簕的果实,通过显微镜观察其横切面和纵切面的显微特征
显微观察露兜簕新鲜果实的徒手切片,易见许多维管束,在中心横切面中分布较多,维管束中可能有很多纤维结构,但在徒手切片下不易观察,因此露兜簕的果肉部分口感较差。
2.经快速压力溶剂萃取,对露兜簕果的基本化学成分的测定
采用快速压力溶剂萃取,用80%的乙醇,70℃,110bar压力,少量、快速提取用于测定露兜簕果的基本成分,经苯酚-硫酸法测定计算得,露兜簕果中多糖的平均含量为27.7%；采用芦丁标样比色法测得露兜簕果中总黄酮的平均含量为1.235%；利用酸性染料比色法测得露兜簕果中总生物碱的平均含量为0.158%(以苦参碱计)。
3.通过乙醇加热回流法提取,经有机溶剂萃取,GC-MS分析各部分的相关有效成分
GC-MS分析四个萃取部分的化学成分,共鉴定出22种化合物,其中石油醚部分鉴定出了16种,氯仿部分13种,乙酸乙酯部分9种,正丁醇部分7种,分别占总峰面积的84.27%,80.20%,73.22%和74.29%,其中脂肪酸类物质最多,占总醇提物60%～70%,在四种萃取物中均发现亚油酸、油酸、硬脂酸、棕榈酸。
4.用GC对露兜簕果中的角鲨烯的含量进行测定
采用超临界CO2提取,并优化了提取工艺。结果显示露兜簕果中含有角鲨烯,但含量较低约为0.02%。
5.利用ICP-MS对生长环境不同的露兜簕果的微量元素等进行分析检测
采用微波消解技术将露兜簕果进行消解处理,再利用ICP-MS测定了两种不同产地的露兜簕果中的Li、B、V、Mn、Cu、As、Sr、Mo、Cs、Ba、Pb和Cr等多种微量元素。结果表明,各元素的检出限为0.001～4.05μg/g,相对标准偏差为0.2%～5.0%,回收率为85%～105%。
6.采用薄层色谱和硅胶柱色谱技术对露兜簕萃取物进行分离纯化
研究表明,硅胶GF254薄层色谱法的最佳展开剂配比为石油醚：乙酸乙酯=8：2,经硅胶柱色谱分离纯化,最后在石油醚段得到纯物质F5-10
7.用IR、13C-NMR、1H-NMR、LC/MS对分离出的纯物质进行结构解析通过对谱图的分析,并结合参考文献,最后确定该物质为齐墩果酸。本课题得到了海南省自然科学基金项目(210010)的经费支持。
Pandanus tectorius Sol. also known as wild pineapple, widely distributed in Asia, Africa and Oceania in tropical and subtropical regions, its roots, leaves, flowers and fruits can be medicinal. In this thesis, wild pineapple fruit on the active ingredients and related research to better analyze potential resource value of the wild pineapple, also provide some basis for the development and utilization of the plant resources and scientific research.The main research contents and results are as follows:
1. Understand the microscopic characteristics of Pandanus tectorius Sol..Observed through microscope cross-section and longitudinal section of the fresh fruit.
Microscopic observation the fresh fruit of Pandanus tectorius Sol. by hand slicing, easy to see that many vascular bundles distributed in the central cross-section. There may be a lot of fiber bundle structure, but it's difficult to observe under the hand section, so the flesh part of the pandanus poor taste.
2. Determine the basic chemical composition in the fruit of Pandanus tectorius Sol. by the rapid pressure solvent extraction.
By rapid pressure solvent extraction, with 80% ethanol,70℃,11Obar pressure, Small, fast extraction for the determination of the basic components of wild pineapple fruit. By the phenol-sulfuric acid method calculated:the average content of polysaccharides is 27.7%; Using Rutin standard samples measured the average content of total flavonoids is 1.235%in the pandanus's fruit; Measured by acid dye colorimetry, the average total alkaloid content of 0.158% (Calculated by Matrine).
3. Extracted by ethanol reflux, the organic solvent extraction, GC-MS analysis the active ingredient.
GC-MS analysis of some of the chemical composition of the four extracts,22 compounds were identified, of which petroleum ether identified 16 species, chloroform13 species, ethyl acetate 9 species, n-butano17 species; respectively,84.27%,80.20%,73.22%and 74.29%of the total peak area. Fatty substances is the most,60% to 70% of the total ethanol extract. And in the four extracts were all found linoleic acid, oleic acid, the hard fatty acid, palmitic acid.
4. Determine squalene content in the fruit of the pandanus with the GC.
The results showed that the pandanus fruit contains squalene, but low levels, about 0.02%.
5. Detected and analyzed the trace elements of the pandanus fruit in different growth conditions by ICP-MS.
Microwave digestion to digest the wild pineapple fruit, and then determined Li, B, V, Mn, Cu, As, Sr, Mo, Cs, Ba, Pb, Cr and other trace elements of two different areas pandanus fruit by ICP-MS. The results showed that the detection limit of each element is 0.001 mg/g to 4.05 mg/g, relative standard deviation is 0.2% to 5.0%, and the recovery is 85% to 105%.
6. Isolated the organic solvents extracts through TLC and silica gel column chromatography.
Studies have shown that the best developing solvent ratio of silica gel GF254 thin-layer chromatography is petroleum ether:ethyl acetate= 8:2, by silica gel column chromatography purified, in the petroleum ether get pure substance F5-1.
7. Analysis the structure of pure substance by IR,13C-NMR,1H-NMR, LC/MS.
Through the spectrum analysis, combined with references to finalize the substance is Oleanolic acid.
This work was supported by The natural science fund of Hainan province (210010).
1.取新鲜露兜簕的果实,通过显微镜观察其横切面和纵切面的显微特征
显微观察露兜簕新鲜果实的徒手切片,易见许多维管束,在中心横切面中分布较多,维管束中可能有很多纤维结构,但在徒手切片下不易观察,因此露兜簕的果肉部分口感较差。
2.经快速压力溶剂萃取,对露兜簕果的基本化学成分的测定
采用快速压力溶剂萃取,用80%的乙醇,70℃,110bar压力,少量、快速提取用于测定露兜簕果的基本成分,经苯酚-硫酸法测定计算得,露兜簕果中多糖的平均含量为27.7%；采用芦丁标样比色法测得露兜簕果中总黄酮的平均含量为1.235%；利用酸性染料比色法测得露兜簕果中总生物碱的平均含量为0.158%(以苦参碱计)。
3.通过乙醇加热回流法提取,经有机溶剂萃取,GC-MS分析各部分的相关有效成分
GC-MS分析四个萃取部分的化学成分,共鉴定出22种化合物,其中石油醚部分鉴定出了16种,氯仿部分13种,乙酸乙酯部分9种,正丁醇部分7种,分别占总峰面积的84.27%,80.20%,73.22%和74.29%,其中脂肪酸类物质最多,占总醇提物60%～70%,在四种萃取物中均发现亚油酸、油酸、硬脂酸、棕榈酸。
4.用GC对露兜簕果中的角鲨烯的含量进行测定
采用超临界CO2提取,并优化了提取工艺。结果显示露兜簕果中含有角鲨烯,但含量较低约为0.02%。
5.利用ICP-MS对生长环境不同的露兜簕果的微量元素等进行分析检测
采用微波消解技术将露兜簕果进行消解处理,再利用ICP-MS测定了两种不同产地的露兜簕果中的Li、B、V、Mn、Cu、As、Sr、Mo、Cs、Ba、Pb和Cr等多种微量元素。结果表明,各元素的检出限为0.001～4.05μg/g,相对标准偏差为0.2%～5.0%,回收率为85%～105%。
6.采用薄层色谱和硅胶柱色谱技术对露兜簕萃取物进行分离纯化
研究表明,硅胶GF254薄层色谱法的最佳展开剂配比为石油醚：乙酸乙酯=8：2,经硅胶柱色谱分离纯化,最后在石油醚段得到纯物质F5-10
7.用IR、13C-NMR、1H-NMR、LC/MS对分离出的纯物质进行结构解析通过对谱图的分析,并结合参考文献,最后确定该物质为齐墩果酸。本课题得到了海南省自然科学基金项目(210010)的经费支持。
Pandanus tectorius Sol. also known as wild pineapple, widely distributed in Asia, Africa and Oceania in tropical and subtropical regions, its roots, leaves, flowers and fruits can be medicinal. In this thesis, wild pineapple fruit on the active ingredients and related research to better analyze potential resource value of the wild pineapple, also provide some basis for the development and utilization of the plant resources and scientific research.The main research contents and results are as follows:
1. Understand the microscopic characteristics of Pandanus tectorius Sol..Observed through microscope cross-section and longitudinal section of the fresh fruit.
Microscopic observation the fresh fruit of Pandanus tectorius Sol. by hand slicing, easy to see that many vascular bundles distributed in the central cross-section. There may be a lot of fiber bundle structure, but it's difficult to observe under the hand section, so the flesh part of the pandanus poor taste.
2. Determine the basic chemical composition in the fruit of Pandanus tectorius Sol. by the rapid pressure solvent extraction.
By rapid pressure solvent extraction, with 80% ethanol,70℃,11Obar pressure, Small, fast extraction for the determination of the basic components of wild pineapple fruit. By the phenol-sulfuric acid method calculated:the average content of polysaccharides is 27.7%; Using Rutin standard samples measured the average content of total flavonoids is 1.235%in the pandanus's fruit; Measured by acid dye colorimetry, the average total alkaloid content of 0.158% (Calculated by Matrine).
3. Extracted by ethanol reflux, the organic solvent extraction, GC-MS analysis the active ingredient.
GC-MS analysis of some of the chemical composition of the four extracts,22 compounds were identified, of which petroleum ether identified 16 species, chloroform13 species, ethyl acetate 9 species, n-butano17 species; respectively,84.27%,80.20%,73.22%and 74.29%of the total peak area. Fatty substances is the most,60% to 70% of the total ethanol extract. And in the four extracts were all found linoleic acid, oleic acid, the hard fatty acid, palmitic acid.
4. Determine squalene content in the fruit of the pandanus with the GC.
The results showed that the pandanus fruit contains squalene, but low levels, about 0.02%.
5. Detected and analyzed the trace elements of the pandanus fruit in different growth conditions by ICP-MS.
Microwave digestion to digest the wild pineapple fruit, and then determined Li, B, V, Mn, Cu, As, Sr, Mo, Cs, Ba, Pb, Cr and other trace elements of two different areas pandanus fruit by ICP-MS. The results showed that the detection limit of each element is 0.001 mg/g to 4.05 mg/g, relative standard deviation is 0.2% to 5.0%, and the recovery is 85% to 105%.
6. Isolated the organic solvents extracts through TLC and silica gel column chromatography.
Studies have shown that the best developing solvent ratio of silica gel GF254 thin-layer chromatography is petroleum ether:ethyl acetate= 8:2, by silica gel column chromatography purified, in the petroleum ether get pure substance F5-1.
7. Analysis the structure of pure substance by IR,13C-NMR,1H-NMR, LC/MS.
Through the spectrum analysis, combined with references to finalize the substance is Oleanolic acid.
This work was supported by The natural science fund of Hainan province (210010).
引文
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