银杏叶、银杏叶提取物口服制剂中黄酮苷类成分质量分析研究

发布时间：2018-01-18 16:18

本文关键词：银杏叶、银杏叶提取物口服制剂中黄酮苷类成分质量分析研究　出处：《北京中医药大学》2017年硕士论文　论文类型：学位论文

【摘要】：银杏叶为裸子植物银杏Ginkgo biloba L.的干燥叶,以银杏叶或银杏叶提取物为原料的制剂是现代植物药的重要产品,占据较大的市场份额,此类产品的质量问题时有报道,解决问题的关键在于提高现行标准的质量控制水平。本论文聚焦银杏叶中主要有效成分之一的黄酮苷类化合物,通过采用指纹图谱、一测多评以及对照提取物等技术手段,研究更加有效、简便和可行的质控方法。由于其组成尚未完全阐明,加之对照物质难以获得,目前国内外现行标准中,对于银杏叶、银杏叶提取物以及一系列制剂中银杏黄酮苷类的质量控制均采用酸水解这一间接方法,经折算后计算出总黄酮苷的含量,该法虽可解决对照物质缺乏的问题,但对质量控制不准确,并为人为添加黄酮苷元等造假行为提供了可能。本论文在前期研究的基础上,对银杏叶制剂中黄酮苷原型化合物,研究建立了银杏叶片、胶囊、酊剂、滴剂、滴丸、软胶囊等口服制剂的指纹图谱,并采用相似度评价和特征黄酮苷峰面积比双质控指标相结合的方式对样品进行定性分析研究,最终确定更加耐用和可行的判定标准,即相似度0.90的同时,特征黄酮苷与芦丁的峰面积比((KRG+QRcG+KRcG)/Rutin)应当不低于1.60。为了解决银杏叶黄酮苷对照品难获得、成本高的问题,本论文在实验室前期研究的基础上,建立了一测多评方法,针对银杏叶中黄酮苷原型化合物,以芦丁为内参物,建立了芦丁与其他5个黄酮苷的相对校正因子,实现了用"一测多评"法测定银杏叶口服制剂中黄酮苷的含量。定向纯化制备了银杏黄酮苷对照提取物,优化制备工艺,从银杏叶提取物中分离制备出含有5个银杏黄酮苷指标性成分的对照提取物,并对黄酮苷含量进行了标定,通过量值转化,可用于测定银杏叶口服制剂中黄酮苷的含量。对中国食品药品检定研究院发放的银杏叶对照提取物(供鉴别用)中6种黄酮苷含量进行标定,并通过量值转移,用于测定银杏叶口服制剂中黄酮苷的含量。这3种方法的测定结果与外标法测定结果比较,统计分析结果显示,方法间无明显差异(P均0.05),说明均可用于银杏叶多剂型口服制剂的定量分析。为了探究银杏叶中黄酮苷类成分积累与季节变化的关系,2016年5月到10月底,对北京市3个不同地区、相同树龄、不同性别的银杏共162份银杏叶样品进行生长周期内黄酮苷类成分含量的动态监测,结果表明不同地区所测成分含量随季节变化的趋势基本一致,即三种黄酮苷芦丁、山柰酚3-O-芸香糖苷、异鼠李素3-O-芸香糖苷含量都是在5月份变化不大,6、7月下降,8月开始回升,之后到10月虽有波动但一直保持较高的水平;其他三种黄酮苷山柰酚3-O-鼠李糖-2-葡萄糖苷、槲皮素3-O-鼠李糖-2-O-(6-O-对羟基反式桂皮酰)-葡萄糖苷、山柰酚3-O-鼠李糖-2-O-(6-O-对羟基反式桂皮酰)-葡萄糖苷含量变化情况基本一致,在整个生长周期内各月虽有波动变化,但整体变化不大,基本保持平稳。雌、雄株中含量存在差异,雄株银杏叶中6种黄酮苷含量高于雌株银杏叶。三个地区雌、雄株银杏叶中六种黄酮苷含量最大值出现的月份稍有不同,其中雌株分别出现在5月、8月、10月,雄株分别出现在5月和10月。上述研究和结果,为银杏叶及其口服制剂的质量控制提供了简便、耐用和可行的新方法,提高了银杏叶及其口服制剂的质量控制水平。
[Abstract]:Ginkgo biloba ginkgo Ginkgo biloba L. for gymnosperms, the dried leaf of Ginkgo biloba preparations or Ginkgo biloba extract as raw materials is an important product of modern plant medicine, occupy a larger market share, the quality problem of such products have been reported, the key to solve the problem is to improve the quality control level of the current standard. Flavonoids one of the main effective components of this paper focus in Ginkgo leaves, through the use of fingerprint QAMs and control extracts and other technical means to study more effective quality control method is simple and feasible. The group has not been fully elucidated, and the control of material is difficult to obtain, at home and abroad in the current standards, for Ginkgo biloba the quality control of Ginkgo biloba extract and a series of preparations of Ginkgo flavone glycosides are using this indirect method by acid hydrolysis, after conversion to calculate the content of total flavonoid glycosides, the Although the method can solve the control problem of lack of material, but the quality control is not accurate, and provides the possibility to artificially add flavonoid aglycones such as fraud. This paper based on the previous studies, the prototype of the flavonoids of Ginkgo biloba extract in the compound, study the establishment of Ginkgo Biloba Leaves Extract Tablets, capsule, tincture, pill drops, fingerprint atlas of soft capsule oral preparation, and the similarity evaluation and characteristics of flavonoid glycosides in peak area than the double quality control index of combination of qualitative analysis and Study on the sample, and ultimately determine the more durable and feasible criteria, namely 0.90 similarity and peak area characteristics of flavonol glycoside and rutin ratio ((KRG+QRcG+KRcG) /Rutin) it should be no less than 1.60. in order to solve the ginkgo flavone glycoside reference is difficult to obtain, the problem of high cost, this paper based on previous studies, the establishment of the QAMs method for Ginkgo biloba leaves The prototype of flavonoid glycosides compounds, with rutin as a reference substance, a relative correction factor of rutin and other 5 flavonoid glycosides, implementation of the "content of a QAMs method for the determination of flavonoid glycosides in Ginkgo biloba oral preparation. The preparation and purification of Ginkgo flavone glycosides extract directional control, optimizing the preparation process, preparation a control extract of Ginkgo flavone glycosides 5 index components isolated from the extract of Ginkgo biloba leaves, and the contents of flavonoids were calibrated, value transformation through, can be used for the determination of Ginkgo biloba in the oral preparations of Huang Tonggan. The Chinese Institute for food and drug control release Ginkgo biloba extract (for identification) in 6 kinds of flavone glycosides were calibrated, and through value transfer, for the determination of flavonoid glycosides in Ginkgo biloba oral preparation. The determination results of the 3 methods external standard method and the results were compared, the results of statistical analysis show that. No significant difference between the method (P 0.05), that can be used for quantitative analysis of Ginkgo biloba multi dosage oral preparation. In order to explore the relationship between flavonoid glycosides accumulation and seasonal variation in Ginkgo biloba leaves, May 2016 to the end of October, the same in Beijing city in 3 different areas, different age, sex and a total of 162 Ginkgo biloba leaf samples were dynamic monitoring during the growth cycle of flavonoid glycosides content, the results show that the different area measured by the composition of seasonal changes in the content of trend is basically the same, namely three kinds of flavonoids rutin, kaempferol rutinoside 3-O-, isorhamnetin 3-O- rutinoside content is little change in May, 6,7 month decline since August again, after October despite fluctuations but has maintained a high level; the other three flavonoid glycosides of kaempferol 3-O- rhamnose glucose -2- glycosides, quercetin 3-O- rhamnose -2-O- (6-O- hydroxy trans cinnamoyl) - glucosidase. Kaempferol, 3-O- rhamnosus -2-O- (6-O- hydroxy trans cinnamoyl) - glucoside content changes are basically the same, in the whole growth period of each month despite fluctuations, but the overall change is not big, remained stable. The female, there are differences in male plants, the content of 6 kinds of male ginkgo flavone glycosides in the leaves of Ginkgo biloba were higher than female. Three in female, the content of six kinds of flavonoid glycosides in Ginkgo biloba leaves male strain maximum month is slightly different, the female plants respectively in May, August, October, male plants respectively in May and October. The above research and results for the quality control of ginkgo leaf and the oral formulation provides a simple and feasible method, new durable, improve the level of quality control of Ginkgo biloba and its oral preparation.

【学位授予单位】：北京中医药大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R286.0

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