【作者】 吴好好；

【导师】 蒋惠娣；

【作者基本信息】 浙江大学 ， 药物分析学， 2006， 硕士

【摘要】 桑叶为桑属植物桑树(Morus alba L.)的树叶。桑叶中营养丰富,含有氨基酸、维生素、矿物质、黄酮、生物碱、糖类、甾类等多种功能成分,具有降血糖、降血脂、降血压、抗炎、抗衰老、抗肿瘤等功效,桑叶在医药和食品领域中得到了广泛的应用。 本文对不同采摘时间、不同品种、不同种植基地的桑叶进行了指纹图谱和黄酮类成分分析以及黄酮分离纯化工艺的研究,通过比较桑叶药材的内在品质和鉴别特征、分离桑叶有效部位,为湖州桑叶开发利用提供依据。此外,对包含部分桑叶黄酮物质在内的黄酮类化合物进行了色谱保留行为和舒血管活性的相关性考察,希望为该类药物的构效研究提供参考。 1.桑叶药材的HPLC指纹图谱研究 目的:建立桑叶药材的高效液相指纹图谱(HPLC-FP),为桑叶药材的鉴别及品质控制提供依据。 方法:采用RP-HPLC法对不同种植基地、不同采摘时间和不同品种的湖州桑叶进行指纹图谱研究,建立HPLC指纹图谱共有模式,并进行聚类分析和相似度比较。 结果:HPLC指纹图谱对桑叶药材中主要成分均得到较好分离,根据聚类分析和相似度分析处理结果,不同种植基地、不同采摘时间和不同品种的桑叶药材指纹图谱相似度较好,但少部分品种的药材指纹图谱有明显差别,从采摘时间看,更多还原

【Abstract】 Folium Mori, leaves of Morus alba L., contain varieties of functional compounds, such as amino acids, vitamins, minerals, flavonoids, alkaloids, carbohydrates, steroids, and so on, with activities of exhibiting a wide range of biological effects, including reducing blood glucose, hypertensive, antihyperlipidemia, anti-inflammatory, antioxidant, anticancer, bacteriostasis and antivirus activity. Folium Mori have been widely used as medicine and food.The present study was to study the HPLC fingerprint, contents determination of flavonoids and the separation and purification of total flavonoids of flavonoids of Folium Mori, so as to provide basis for the development of Folium Mori resource. Moreover, the vasodilator activity of Folium Mori extract and the relationship between vasodilation of flavonoids and their retention index (RI) were also investigated.1. Determination of chromatographic fingerprint of Folium Mori by HPLCObjective: To develop the fingerprint determination method for Folium Mori, by HPLC method and to provide basis for identification of Folium Mori.Methods: A RP-HPLC method with gradient elution was used to determine the fingerprints of Folium Mori, from different plant sites, collected time and varieties. Mutual mode was developed, cluster analysis and similarity calculation were taken for comparison of the samples.Results: Samples from different plant sites, collection time and varieties were of high similarity, though a few samples showed evident difference in fingerprint graphics. Samples harvested in winter had even higher similarity than that of other seasons.Conclusion: Fingerprint of Folium Mori, can be an identifying method for Folium Mori.2. Determination of main flavonoids components of Folium MoriObjective: To determine and compare the contents of total flavonoids, chlorogenic acid, rutin, isoquercitrin, quercetin and kaempferol in Folium Mori from different collected time, varieties and plant sites.Method: The contents of total flavonoids was determined by colorimetry.The contents of chlorogenic acid, rutin and isoquercitrin were determined by RP-HPLC. Zorbax SB Cis column(4.6mm><250mm,5um) was used as analysis column, the mobile phase was 90%acetonitrile-0.2%phosphoric acid with gradient elution. The flow rate was 1.0 mL-min"1 , the column temprature and the detection wavelength were set at 25°C,358 nm. The contents of quercetin and kaempferol were determined by another HPLC method, performed on Zorbax SB Ci8 column with the mobile phase consisted methanol-0.20% phosphoric acid solution (55:45, v/v). The flow rate was 1.0 mL-min’1, the column temprature and UV wavelengh were set at 35°C,370nm. Result: The contents of tested components in Folium Mori samples harvested in winter are higher than that of other seasons., while the contents varied obviously among Folium Mori from different plant sites and different varieties.Conclusion: The contents tested components are influenced by collected time, plane site, and varieties.3. Studies on the separation and purification of total flavonoids from Folium Mori, by macroreticular resinObjective: To investigate the technological parameters of the purification process of total flavonoids from Folium Mori, by macroreticular resins.Method: The static absorption capacity,static elution ratio, static elution capacity of twelve types macroreticular resins were studied respectively and compared in order to find the optimum macroreticular resin. The technical process for purification of total flavonoids with the optimum macroreticular resin, was screened by purity and yield of total flavonoids product.Result: Rl type macroreticular resin owned optimum absorption and eluationparameters.The absorption ratio was 111.1 mgg"* . After the optimization, the yield of total flavonois was 76.6% and product purity was 56.0%.Conclusion: The Rl type macroreticular resin showed better comprehensive absorption property. It can be used to purify the extract of Folium Mori..4. Studies on vasodilator activity of Folium Mori extract and the relationship between vasodilation of flavonoids and their retention indexObjective: To investigate the vasodilator activity of Folium Mori extract and the relationship between vasodilation of flavonoids and their retention index (RI).Methods: Vasorelaxation of the flavonoids and Folium Mori extract were measured by measuring the tension reduction by samples in rat thoracic aorta precontracted by phenylephrine (PE) or by KC1. RI values of 18 flavonoids, including flavones, flavonois, flavanones, flavanols and isoflavones were measured by RP-HPLC method based on alkan-2-ones.Results: The RI-Rex regression equation for flavonoids was Rex =-11.68+0.11 RI (n=18,r =0.915, PO.01). Both water extract and ethanol extract caused concentration-dependent relaxation of aorta rings precontricted with K+(P<0.01), vasodilator effect of ethanol extract is greater than that of water extract(P<0.01).Conclusion: Folium Mori extract induces relaxation of aorta rings precontricted with K+.The potency of the vasorelaxation of flavonoids was positive correlated with RI.更多还原