青叶胆的化学成分研究及2010版《中国药典》青叶胆标准修订研究
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摘要
青叶胆为龙胆科植物青叶胆Swertia mileensis T.N.Ho etW.L.Shih的干燥全草,味苦,微寒。主要分布于云南弥勒、开远等县,具有清肝利胆,清热利湿的功效,用于黄疸尿赤,热淋涩痛,为《中国药典》收载品种。
青叶胆化学成分众多,主要含有黄酮类、环烯醚萜苷类、(口山)酮类以及三萜类化合物。文献报道其有效成分具有保肝、解痉、镇痛、抗炎、抗病毒等药理作用。本实验将青叶胆药材晒干切断,用95%EtOH浸提,回收溶剂后,分散于水中,分别用氯仿、水饱和正丁醇依次萃取。采用硅胶柱层析、葡聚糖凝胶LH-20柱层析等分离方法对正丁醇部分所含成分进行分离,得到了4个单体化合物。通过理化常数测定及光谱分析(UV、IR、~(13)CNMR、~1HNMR)等方法鉴定了其结构,分别为:獐牙菜苦苷、龙胆苦苷、齐墩果酸和异牡荆素。对青叶胆中化学成分的分离及鉴定,为獐牙菜属植物的化学分类学,属间亲缘关系的相关研究以及合理开发利用青叶胆植物资源提供了依据。
在获得上述化学成分的基础上,本实验考察了青叶胆中獐牙菜苦苷及异牡荆素在高效毛细管电泳中的迁移行为,建立了毛细管电泳法快速分离分析以上两种成分的方法。通过各种条件的优化,发现背景缓冲液的pH值、缓冲液的浓度、分离电压等因素对分离均有重要影响。该方法灵敏度高,快速、稳定性好:相对于HPLC,具有分离速度快,样品耗量少,环境友好以及成本低等特点,可以作为控制青叶胆药材质量的快速、有效的分析方法。
本专题同时承担了《中华人民共和国药典》2010版(Ⅰ部)标准研究课题:青叶胆药材及饮片质量标准修订。作者采用HPLC法,考察了不同产地青叶胆药材及饮片中獐牙菜苦苷及龙胆苦苷的含量,制定了其含量限度,并对饮片进行了性状、显微、薄层等指标的鉴别研究,测定了药材及饮片中水分、灰分、酸不溶性灰分的含量,这些工作对于评价药材质量、充分利用药物资源具有重要意义。以上研究结果,云南省食品药品检验所正在开展复核检验工作。
Mile Swertia Herb, a traditional Chinese drug "Qingyedan", is the dried herb of Swertia mileensis T. N. Hoet W. L. Shih, which belongs to the Fam. Gentianaceae. Qingyedan is bitter in taste and slight cold in nature. The drug mainly distributes in the Yunnan Mile and Kaiyuan County, et al.. It can increase the flow of bile, and remove heatdampness. Collected in Chinese pharmacopeia, the drug is used for the treatment of tanudice with dark urine, acute urinary infetion with difficult painful urination.
Flavonoids, iridoids, xanthones and triterpenoids are considered as the predominant ingredients in Mile Swertia Herb. In many related reports, the effective components have considerable activities such as liver protection, spasmolysis, analgesia, anti-inflammatory, anti-virus, etc.. In this research, Mile Swertia Herb was collected, cut into pieces and dried. Then the drug was immersed with 95% ethanol, after removing the solvent, the ethanol extracts were dispersed into water, extracted with dichlormethane and n-BuOH saturated with water in sequence. The n-BuOH extracts were subjected to silica gel column chromatograph and sephadex LH-20 column chromatograph to obtain four compounds. Through measuring their physical and chemical constants and spectroscopic analysis (UV、IR、~(13)CNMR、~1HNMR) , four of the structures were elucidated as swertiamarin , gentiopieroside , oleanolic acid and isovitexin. The isolation and identification researches into chemical constituents of Mile Swertia Herb have provided bases on chemosystematics of species swertia herb, the relevant study of genetic relationship and utilization of plant resources.
On the basis of obtaining the components mentioned obove, the author developed a quick and accurate capillary electrophoresis method for the quantitative analysis of the compuonds by studying the migration behavior of swertiamarin and isovitexin on high performance capillary electrophoresis (HPCE). According optimising all the factors involved, we found that the power of hydrogen, concentration of buffer solution and the separation voltage have a significant impact on separate effect. The method is highly sensitive and rapid with good stability. Compared to HPLC, HPCE has the quality of satisfactory separation speed, less sample consumption, environment friendly and low cost. It can sever as a rapid and effective analytical method for the quality control of Mile Swertia Herb.
Meanwhile, we assumed one standard research project of Pharmacopoeia of People's Republic of China Edition 2010: Quality specification revision of Mile Swertia Herb. In this part, HPLC method was adopted to determine the contents of swertiamarin and gentiopieroside in medical material and decoction pieces of Mile Swertia Herb of different habits. Simultaneously, the content limitation was established. In addition, the description, microstructure, TLC, water content, ash content and acid-insoluble ash content of Mile Swertia Herb were researched. All these studies have shown significant importance to evaluate the quality and sufficiently utilize the resource of Mile Swertia Herb. The check of the experimental results now is being carried out by Yunnan Institute For Food and Drug Control.
青叶胆化学成分众多,主要含有黄酮类、环烯醚萜苷类、(口山)酮类以及三萜类化合物。文献报道其有效成分具有保肝、解痉、镇痛、抗炎、抗病毒等药理作用。本实验将青叶胆药材晒干切断,用95%EtOH浸提,回收溶剂后,分散于水中,分别用氯仿、水饱和正丁醇依次萃取。采用硅胶柱层析、葡聚糖凝胶LH-20柱层析等分离方法对正丁醇部分所含成分进行分离,得到了4个单体化合物。通过理化常数测定及光谱分析(UV、IR、~(13)CNMR、~1HNMR)等方法鉴定了其结构,分别为:獐牙菜苦苷、龙胆苦苷、齐墩果酸和异牡荆素。对青叶胆中化学成分的分离及鉴定,为獐牙菜属植物的化学分类学,属间亲缘关系的相关研究以及合理开发利用青叶胆植物资源提供了依据。
在获得上述化学成分的基础上,本实验考察了青叶胆中獐牙菜苦苷及异牡荆素在高效毛细管电泳中的迁移行为,建立了毛细管电泳法快速分离分析以上两种成分的方法。通过各种条件的优化,发现背景缓冲液的pH值、缓冲液的浓度、分离电压等因素对分离均有重要影响。该方法灵敏度高,快速、稳定性好:相对于HPLC,具有分离速度快,样品耗量少,环境友好以及成本低等特点,可以作为控制青叶胆药材质量的快速、有效的分析方法。
本专题同时承担了《中华人民共和国药典》2010版(Ⅰ部)标准研究课题:青叶胆药材及饮片质量标准修订。作者采用HPLC法,考察了不同产地青叶胆药材及饮片中獐牙菜苦苷及龙胆苦苷的含量,制定了其含量限度,并对饮片进行了性状、显微、薄层等指标的鉴别研究,测定了药材及饮片中水分、灰分、酸不溶性灰分的含量,这些工作对于评价药材质量、充分利用药物资源具有重要意义。以上研究结果,云南省食品药品检验所正在开展复核检验工作。
Mile Swertia Herb, a traditional Chinese drug "Qingyedan", is the dried herb of Swertia mileensis T. N. Hoet W. L. Shih, which belongs to the Fam. Gentianaceae. Qingyedan is bitter in taste and slight cold in nature. The drug mainly distributes in the Yunnan Mile and Kaiyuan County, et al.. It can increase the flow of bile, and remove heatdampness. Collected in Chinese pharmacopeia, the drug is used for the treatment of tanudice with dark urine, acute urinary infetion with difficult painful urination.
Flavonoids, iridoids, xanthones and triterpenoids are considered as the predominant ingredients in Mile Swertia Herb. In many related reports, the effective components have considerable activities such as liver protection, spasmolysis, analgesia, anti-inflammatory, anti-virus, etc.. In this research, Mile Swertia Herb was collected, cut into pieces and dried. Then the drug was immersed with 95% ethanol, after removing the solvent, the ethanol extracts were dispersed into water, extracted with dichlormethane and n-BuOH saturated with water in sequence. The n-BuOH extracts were subjected to silica gel column chromatograph and sephadex LH-20 column chromatograph to obtain four compounds. Through measuring their physical and chemical constants and spectroscopic analysis (UV、IR、~(13)CNMR、~1HNMR) , four of the structures were elucidated as swertiamarin , gentiopieroside , oleanolic acid and isovitexin. The isolation and identification researches into chemical constituents of Mile Swertia Herb have provided bases on chemosystematics of species swertia herb, the relevant study of genetic relationship and utilization of plant resources.
On the basis of obtaining the components mentioned obove, the author developed a quick and accurate capillary electrophoresis method for the quantitative analysis of the compuonds by studying the migration behavior of swertiamarin and isovitexin on high performance capillary electrophoresis (HPCE). According optimising all the factors involved, we found that the power of hydrogen, concentration of buffer solution and the separation voltage have a significant impact on separate effect. The method is highly sensitive and rapid with good stability. Compared to HPLC, HPCE has the quality of satisfactory separation speed, less sample consumption, environment friendly and low cost. It can sever as a rapid and effective analytical method for the quality control of Mile Swertia Herb.
Meanwhile, we assumed one standard research project of Pharmacopoeia of People's Republic of China Edition 2010: Quality specification revision of Mile Swertia Herb. In this part, HPLC method was adopted to determine the contents of swertiamarin and gentiopieroside in medical material and decoction pieces of Mile Swertia Herb of different habits. Simultaneously, the content limitation was established. In addition, the description, microstructure, TLC, water content, ash content and acid-insoluble ash content of Mile Swertia Herb were researched. All these studies have shown significant importance to evaluate the quality and sufficiently utilize the resource of Mile Swertia Herb. The check of the experimental results now is being carried out by Yunnan Institute For Food and Drug Control.
引文
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[4]Neerja Pant,DC Jain,RS Bhakuni.Phytochemicals from genus Swertia and their biological activities[J].Indian Journal of Chemistry.2000,39B:565-586
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[8]郭爱华.龙胆科獐牙菜属药用植物化学成分和药理作用的研究进展[J].山西中医学院学报.2005,6(1):57-59
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