蓼实种子提取物的分离、纯化以及结构测定的研究
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摘要
水蓼(Polygonum hydropiper L．)属蓼科(Polygonaceae)蓼属(Polygonum L．)的一年生草本植物，是一种资源十分丰富的植物，在中国大部分地区都有分布。水蓼在植物源农药、医药、兽药、食品添加剂等方面有着良好的开发前景。论文在大量文献的基础上，对蓼实种子中的黄酮类化合物、脂溶性部位化学成分及挥发油化学成分进行了提取、分析、分离、纯化研究，通过HPLC、GC-MS等仪器手段进行定性、定量分析并初步确定了其结构。为进一步开发利用蓼实种子提供了实验依据与基础。共进行了三个方面的研究工作：
1、蓼实种子总黄酮的提取及纯化。蓼实种子总黄酮提取的最佳条件为：溶剂为70％的乙醇；温度85℃；料液比1：15；提取次数2次，回流浸提2.0h。分别用石油醚萃取法、聚酰胺吸附法和活性炭吸附法对蓼实种子中总黄酮进行了纯化工艺研究，得到总黄酮的纯度分别为79.2％、91.8％和78.1％，说明以聚酰胺树脂纯化效果最好。
2、蓼实种子总黄酮的分析鉴定。用显色反应对蓼实种子总黄酮进行了定性分析，初步确定了蓼实种子总黄酮含有黄酮、二氢黄酮、二氢黄酮醇；用紫外分光光度法测得总黄酮的含量为4.32％，该方法的重现性、精密度、稳定性和回收率均较好，分析结果可靠；用高效液相色谱法(HPLC)测定蓼实种子总黄酮含量为4.35％。
3、蓼实种子中挥发性化学成分分析。分别运用水蒸汽蒸馏法和溶剂提取法提取了蓼实种子中挥发性化学成分，并进行了GC-MS分析。通过G170LBA化学工作站数据处理系统，检索NIST147谱图库，并分别与标准谱图进行对照、复合，再结合有关文献进行人工谱图解析及气相色谱保留指数法数据对比，确认了蓼实种子中的各种挥发性化学成分。通过化学工作站数据处理系统，并运用峰面积归一化法进行数据处理，求得各化学成分的相对百分含量。水蒸汽蒸馏提取的挥发油共鉴定出44种化合物，含量相对较高的为β-雪松烯(含量为10.76％)，葎草烯(含量为11.60％)，十六烷酸(含量为16.74％)，(Z)-9-十六碳烯醛(含量为15.39％)，8-溴-新长叶烯(4.61％)。溶剂提取法提取的挥发油共鉴定出41种化合物，其中α-蒎烯的含量高达46.74％。
Polygonum hydropiper L, one of the genus Polygonum, is an annual herb which is abundant in most area of China. It has an extensive application prospect in the fields of pesticides, medicine, veterinary medicine as well as food additives. This paper was concerned with the total flavones, liposoluble constituents and volatile oil from the seeds of Polygonum hydropiper L based on a large number of literature investigation and literature systematization. The ingredients were extracted、purified、analyzed and characterization. The qualitative and quantitative analysis was carried out by means of HPLC, GC-MS to determine the structure of the monomers. The following researches have been carried out:
1. The extraction and purification of the flavonoids. The optimum extraction condition was established as follows: solvent 70% (v/v) ethanol, extraction temperature 85℃, and ratio of material to liquid(g/mL) 1:15. extraction time 2.0h, extraction times twice. The purification methods of petroleum ether extraction, polyamide adsorption and active carbon adsorption resin were studied systemically. The polyamide adsorption resin was chosen for its higher purity. By the way, the percentage purity of above three methods were 79.2%, 91.8% and 78.1%.
2. The analysis and identification of the flavonoids. The qualitative analysis of the flavonoids by silica gel chromatogram and colouring showed that the extraction solution contains flavonoids. The content of the flavonoids from the seeds of Polygonum hydropiper L. was analyzed quantitatively by spectrophotometry and effect of parameters including degree of precision, repeatability, stability, recovery of the samples on measuring result was investigated, and the content was 4.32%. The content of the flavonoids were also analyzed by HPLC and the result was 4.35%.
3. Analysis of the volatile oil. Both steam-distillation and solvent extraction were used, the compunds were identified by gas chromatography-mass spectrometry(GC-MS) analysis. Analysis was performed by G170LBA chemistry workstation, similarity of the components were searched in NIST147 library and relative contents of volatile components were determined by peak area normalization and then compared with the standardspectrogram. 44 compounds were identified extracted by steamdistillation and the main component were beta.-Himachalene whose percent was 10.76%, Humulen-(v1) whose percent was 11.60%, n-Hexadecanoic acid whose percent was 16.74% , (Z)-9-Hexadecenal whose percent was 15.39% and 8-bromo-Neoisolongifolene whose percent was 4.61%. 41 compounds were identified extracted by solvent extraction within whichα-pinene was as high as 46.74%.
1、蓼实种子总黄酮的提取及纯化。蓼实种子总黄酮提取的最佳条件为：溶剂为70％的乙醇；温度85℃；料液比1：15；提取次数2次，回流浸提2.0h。分别用石油醚萃取法、聚酰胺吸附法和活性炭吸附法对蓼实种子中总黄酮进行了纯化工艺研究，得到总黄酮的纯度分别为79.2％、91.8％和78.1％，说明以聚酰胺树脂纯化效果最好。
2、蓼实种子总黄酮的分析鉴定。用显色反应对蓼实种子总黄酮进行了定性分析，初步确定了蓼实种子总黄酮含有黄酮、二氢黄酮、二氢黄酮醇；用紫外分光光度法测得总黄酮的含量为4.32％，该方法的重现性、精密度、稳定性和回收率均较好，分析结果可靠；用高效液相色谱法(HPLC)测定蓼实种子总黄酮含量为4.35％。
3、蓼实种子中挥发性化学成分分析。分别运用水蒸汽蒸馏法和溶剂提取法提取了蓼实种子中挥发性化学成分，并进行了GC-MS分析。通过G170LBA化学工作站数据处理系统，检索NIST147谱图库，并分别与标准谱图进行对照、复合，再结合有关文献进行人工谱图解析及气相色谱保留指数法数据对比，确认了蓼实种子中的各种挥发性化学成分。通过化学工作站数据处理系统，并运用峰面积归一化法进行数据处理，求得各化学成分的相对百分含量。水蒸汽蒸馏提取的挥发油共鉴定出44种化合物，含量相对较高的为β-雪松烯(含量为10.76％)，葎草烯(含量为11.60％)，十六烷酸(含量为16.74％)，(Z)-9-十六碳烯醛(含量为15.39％)，8-溴-新长叶烯(4.61％)。溶剂提取法提取的挥发油共鉴定出41种化合物，其中α-蒎烯的含量高达46.74％。
Polygonum hydropiper L, one of the genus Polygonum, is an annual herb which is abundant in most area of China. It has an extensive application prospect in the fields of pesticides, medicine, veterinary medicine as well as food additives. This paper was concerned with the total flavones, liposoluble constituents and volatile oil from the seeds of Polygonum hydropiper L based on a large number of literature investigation and literature systematization. The ingredients were extracted、purified、analyzed and characterization. The qualitative and quantitative analysis was carried out by means of HPLC, GC-MS to determine the structure of the monomers. The following researches have been carried out:
1. The extraction and purification of the flavonoids. The optimum extraction condition was established as follows: solvent 70% (v/v) ethanol, extraction temperature 85℃, and ratio of material to liquid(g/mL) 1:15. extraction time 2.0h, extraction times twice. The purification methods of petroleum ether extraction, polyamide adsorption and active carbon adsorption resin were studied systemically. The polyamide adsorption resin was chosen for its higher purity. By the way, the percentage purity of above three methods were 79.2%, 91.8% and 78.1%.
2. The analysis and identification of the flavonoids. The qualitative analysis of the flavonoids by silica gel chromatogram and colouring showed that the extraction solution contains flavonoids. The content of the flavonoids from the seeds of Polygonum hydropiper L. was analyzed quantitatively by spectrophotometry and effect of parameters including degree of precision, repeatability, stability, recovery of the samples on measuring result was investigated, and the content was 4.32%. The content of the flavonoids were also analyzed by HPLC and the result was 4.35%.
3. Analysis of the volatile oil. Both steam-distillation and solvent extraction were used, the compunds were identified by gas chromatography-mass spectrometry(GC-MS) analysis. Analysis was performed by G170LBA chemistry workstation, similarity of the components were searched in NIST147 library and relative contents of volatile components were determined by peak area normalization and then compared with the standardspectrogram. 44 compounds were identified extracted by steamdistillation and the main component were beta.-Himachalene whose percent was 10.76%, Humulen-(v1) whose percent was 11.60%, n-Hexadecanoic acid whose percent was 16.74% , (Z)-9-Hexadecenal whose percent was 15.39% and 8-bromo-Neoisolongifolene whose percent was 4.61%. 41 compounds were identified extracted by solvent extraction within whichα-pinene was as high as 46.74%.
引文
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