【作者】 李安琪；

【导师】 陈玉琼；

【作者基本信息】 华中农业大学 ， 茶学， 2007， 硕士

【摘要】 藤茶(Ampelopsis grossedentata W．T．Wang)是葡萄科蛇葡萄属的一种藤本植物，广泛分布于我国长江流域以南等省区，是我国民间一茶用、药用植物，富含二氢杨梅素等黄酮类物质。我国壮族和瑶族人民将其幼嫩茎叶制成保健茶，用于治疗感冒发热、咽喉肿痛、黄疸型肝炎等症。有关藤茶的研究主要集中在藤茶活性成分的分离鉴定及黄酮的功能活性等方面。本文首次探讨了施肥条件对藤茶黄酮等主要品质成分的影响；采用二次正交旋转组合设计法研究了藤茶主要活性成分黄酮的提取条件；筛选了利用大孔吸附树脂纯化藤茶黄酮的分离纯化条件以及葡聚糖凝胶对藤茶黄酮的纯化效果，以期为藤茶的开发应用提供理论基础。1．施肥条件对藤茶品质成分的影响采用田间随机区组试验设计，研究了氮肥、磷肥、钾肥、复合肥、农家肥5种肥料对藤茶黄酮等主要品质成分的影响。结果表明，不同类型的肥料对藤茶中黄酮、多酚类物质、氨基酸、可溶性糖、可溶性蛋白质、水浸出物以及矿质元素的含量的影响均达到显著水平。施磷、钾肥的藤茶中黄酮、多酚、可溶性糖及水浸出物的含量增加显著，Ca、Mg、Cu、Zn等矿质元素也显著提高。施用氮肥有利于藤茶中可溶性蛋白质、氨基酸及水浸出物含量的提高。复合肥有利于藤茶中Ca、Mg、Zn、Fe等矿质元素的积累。2．黄酮类化合物的提取条件的优化在单因素试验的基础上，采用二次回归正交旋转组合设计方法研究了温度、时间、料液比、乙醇浓度对藤茶黄酮提取率的影响。结果表明，温度、时间、料液比极显著影响藤茶黄酮的提取率，建立的回归模型显著性检验达极显著水平，回归模型的预测值与实测值能较好地拟合。确定藤茶黄酮的最佳浸提条件：提取温度为77.5℃，乙醇浓度65％，料液比1：40，浸提时间90min。3．黄酮类化合物的分离纯化工艺的优化选用11种大孔吸附树脂对藤茶黄酮进行了静态吸附与解吸附试验。结果表明，1号树脂适宜于藤茶黄酮的分离纯化，对黄酮的吸附量为41.12mg／g，解吸率为92.12％。通过动态吸附试验对1号树脂有关参数进行了优化。结果表明，黄酮上样浓度6—7mg／ml，上样体积为柱体积的1／3，流速1.8ml／min，用95％的乙醇进行洗脱，可达到较好的洗脱与纯化效果，黄酮的洗脱率可达90％左右，洗脱出的黄酮纯度达65％左右。进一步利用Sephadex LH-20层析柱进行分离纯化，制得的黄酮纯度达74.40％，二氢杨梅素含量达67.12％。更多还原

【Abstract】 Tengcha(Ampelopsis grossedentata W.T.Wang)belongs to Ampelopsis Michx of Vitaceae. It is grown abroad in the south of the Yangtze River. It’s used for drinking and drug among the people, which contains rich flavones, such as Dihydromyricetin (DMY). The tender stems and leaves of Tengcha have been made into a healthy drink which is called "mystic tea" for treating rheum, fever, gall of fauces, hepatitis and so on, with a long history of several hundred years among Yao and Zhuang nationality in China.Much attention on Tengcha focused on identification of active constituents and biological activities of flavones in recent years. In this study, the effects of fertilization for Tengcha’s main biochemical components such as flavones were studied. Meanwhile, the conditions of extraction by means of quaternion-square regression-rotation orthogonal experimental design, and purification with resins and sephadex LH-20 were reported.1. The effect of fertilization for Tengcha’s qualityWith the stochastic area experiment in the fields design, the effects of 5 kinds of fertilizers (nitrogen fertilizer, phosphate fertilizer, potassium fertilizer, compound fertilizer and farmyard manure) on the contents of Tengcha’s main biochemistry components such as flavones were studied. The results showed that the different kinds of fertilizers had significant effects on the contents of flavone, polyphenols, amino acids, soluble sugar, soluble protein, water extraction and mineral elements of Tengcha. The contents of flavones, polyphenols, soluble sugar, water extraction and elements of Ca、Mg、Cu、Zn of Tengcha fertilized with phosphate fertilizer and potassium fertilizer were more than the control’s and others’. Nitrogen fertilizer was propitious to increase the contents of soluble protein, amino acids and water extraction of Tengcha. In addition, compound fertilizer was good to accumulate the contents of mineral elements Ca、Mg、Cu、Zn.2. The extraction conditions of flavones on TengchaThe effects of extracting temperature, time, the ratio of Tengcha and water, ethanol consistency on the extracting rates of flavones were studied. The results showed that the factors of temperature, time, the ratio of Tengcha and water had significant effects on extractions of flavones. The model of regression of flavones’ extracting rate was statistically significant at 1％level and was appropriate to express response variables. Considering high extracting rate, the better variable levels were 65％ethanol consistency, 77.5℃temperature, 90min heating time, 1:40 ratio of Tengcha and water. 3. Purification of flavones on TengchaExperiments of 11 kinds of macroporous adsorbing resins adsorbing and desorbing flavones indicated that NO. 1 was one of the most appropriate resins in all for preliminary purification of flavones. Its adsorbing capacity was 41.12mg/g, desorbing rate was 92.12％. In the column of NO. 1, the optimum conditions were that the concentration of crude flavones was 6-7mg/ml, the volume of sample was 1/3 that of column, flowing velocity was 1.8ml/min, eluting solution was 95％ethanol, which led to 65％purity of the flavones and 90％desorbing rate of flavones. The purity of flavones with further purification by Sephadex LH-20 was 74.40％and the content of DMY was 67.12％.更多还原