利用HPLC和HPLC/MS分析比较当归有效成分的研究
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摘要
中草药当归[Angelica sinensis (Oliv.) Diels ]为伞形科植物 (Umbelliferae)植物当归的干燥根。当归是著名中药，素有“十方九归”之说，现代药理和临床研究也表明当归对多种疾病特别是心血管系统疾病，抗肿瘤，改善肺功能，提高免疫力有很好的疗效。当归作为药用价值很高的传统中药，也面临着诸多问题。其本身所含的化学成分非常复杂，而且由于其产地的分布十分广泛，不同产地的当归的化学成分及其含量也存在着不同程度的差别，分析工作者难以对当归的化学成分或者药材的质量作出系统的评价。
本文首先采用薄层色谱的方法对甘肃岷县和湖北兴山所产的当归乙醇提取液进行了粗略的比较，通过检视其展开斑点发现两者之间存在着一定的差别。然后用反相高效液相色谱对两种当归乙醇提取物的甲醇溶液采用梯度洗脱进行了分离，并比较了两者之间的异同之处。比较发现两者的主要色谱峰基本相同，但是仍然存在着一定的差别。此外，对岷县当归的头身尾的乙醇提取液甲醇溶液进行了液相分离比较，发现其成分在含量上存在着显著的差别。
作为进一步的研究，本研究采用液质联用技术（HPLC/MS/MS）对岷县当归提取液进行了分析，分别得到了其二极管阵列色谱图，正离子总离子流图，负离子总离子流图及总离子流图。结合已知的当归中有效成分的信息，从质谱图中找到了m/z为191.25正离子分子离子峰和117.08，255.42，193.18的负离子分子离子峰，可能分别为藁本内酯，琥珀酸，棕榈酸，阿魏酸的分子离子峰，但是要确认这些成分还有待深入研究。此外，针对试验中发现的一些现象也提出了相应的解释。
Angelica Sinensis (Oliv.) diels ( Danggui in Chinese) is the root of Umbellifarae plant. Angelica Sinensis is one of the famous Chinese medicinal plants widely used among Chinese traditional medicine prescriptions. Modern pharmacology and clinical research also prove that Angelica Sinensis is effective in anti-ulcer, anti-tumor, improving blood circulation, lung function and immunity, etc. As Angelica Sinensis is valuable phamarcologically, it also faces many problems. Just like most herbs, it contains number of constituents, various in polarity and molecular weight. What's more, it has wide production areas over the nation and because of their locations, the chemical compounds and the contents of them they contain are also different. So, it is hard for analysts to make a unified conclusion to Angelica Sinensis's constituents and a scientific evaluation system to control the quality of commodious Angelica Sinensis.
In this dissertation, research of the chemical constituents of Angelica Sinensis was summarily reviewed, including the essential oil, nonvolatile constituents and the main active components. The phamarcological research situation is also stated.
At first, the comparison between the ethanol extracts from Minxian Danggui and Xingshan Danggui, was simply performed by TLC. The results show that there is difference between the Danggui from the two different locations through the inspection of the separated spots. Then the ethanol extracts from Minxian Danggui and Xingshan Danggui were comparatively analyzed by HPLC through an optimized gradient elution program. Results indicate that the main peaks in the chromatograms are similar, but there are still minor differences that can not be neglected. In addition, the extracts of the three different parts: the head, the main body and the end of Angelica Sinensis from Minxian, were analyzed in the same way. It finds that the contents of constituents in different parts are obviously different.
To further the study, the extracts of Minxian Danggui were analyzed by High pressure Liquid chromatography-electrospray mass spectrometry(HPLC/ESI/MS).
The DAD chromatogram and the total ion chromatograms were obtained from the analysis. According to the information known of the phamarceutical components, the author tries to explain the m/z +191.25 and -117.08，-255.42，-193.18 ion chromatograms, and they perhaps belong to the compounds: ligustilide, succinic acid, ferulic acid, palmitic acid and n-butylide phthalide respectively( to determine those constituents needs further study). Additionally, the problems or facts found during the analysis were explained tentatively.
本文首先采用薄层色谱的方法对甘肃岷县和湖北兴山所产的当归乙醇提取液进行了粗略的比较，通过检视其展开斑点发现两者之间存在着一定的差别。然后用反相高效液相色谱对两种当归乙醇提取物的甲醇溶液采用梯度洗脱进行了分离，并比较了两者之间的异同之处。比较发现两者的主要色谱峰基本相同，但是仍然存在着一定的差别。此外，对岷县当归的头身尾的乙醇提取液甲醇溶液进行了液相分离比较，发现其成分在含量上存在着显著的差别。
作为进一步的研究，本研究采用液质联用技术（HPLC/MS/MS）对岷县当归提取液进行了分析，分别得到了其二极管阵列色谱图，正离子总离子流图，负离子总离子流图及总离子流图。结合已知的当归中有效成分的信息，从质谱图中找到了m/z为191.25正离子分子离子峰和117.08，255.42，193.18的负离子分子离子峰，可能分别为藁本内酯，琥珀酸，棕榈酸，阿魏酸的分子离子峰，但是要确认这些成分还有待深入研究。此外，针对试验中发现的一些现象也提出了相应的解释。
Angelica Sinensis (Oliv.) diels ( Danggui in Chinese) is the root of Umbellifarae plant. Angelica Sinensis is one of the famous Chinese medicinal plants widely used among Chinese traditional medicine prescriptions. Modern pharmacology and clinical research also prove that Angelica Sinensis is effective in anti-ulcer, anti-tumor, improving blood circulation, lung function and immunity, etc. As Angelica Sinensis is valuable phamarcologically, it also faces many problems. Just like most herbs, it contains number of constituents, various in polarity and molecular weight. What's more, it has wide production areas over the nation and because of their locations, the chemical compounds and the contents of them they contain are also different. So, it is hard for analysts to make a unified conclusion to Angelica Sinensis's constituents and a scientific evaluation system to control the quality of commodious Angelica Sinensis.
In this dissertation, research of the chemical constituents of Angelica Sinensis was summarily reviewed, including the essential oil, nonvolatile constituents and the main active components. The phamarcological research situation is also stated.
At first, the comparison between the ethanol extracts from Minxian Danggui and Xingshan Danggui, was simply performed by TLC. The results show that there is difference between the Danggui from the two different locations through the inspection of the separated spots. Then the ethanol extracts from Minxian Danggui and Xingshan Danggui were comparatively analyzed by HPLC through an optimized gradient elution program. Results indicate that the main peaks in the chromatograms are similar, but there are still minor differences that can not be neglected. In addition, the extracts of the three different parts: the head, the main body and the end of Angelica Sinensis from Minxian, were analyzed in the same way. It finds that the contents of constituents in different parts are obviously different.
To further the study, the extracts of Minxian Danggui were analyzed by High pressure Liquid chromatography-electrospray mass spectrometry(HPLC/ESI/MS).
The DAD chromatogram and the total ion chromatograms were obtained from the analysis. According to the information known of the phamarceutical components, the author tries to explain the m/z +191.25 and -117.08，-255.42，-193.18 ion chromatograms, and they perhaps belong to the compounds: ligustilide, succinic acid, ferulic acid, palmitic acid and n-butylide phthalide respectively( to determine those constituents needs further study). Additionally, the problems or facts found during the analysis were explained tentatively.
引文
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