金钱草中总黄酮提取工艺的比较研究.doc

金钱草中总黄酮提取工艺的比较研究

摘    要
金钱草系报春花科珍珠菜属植物过路黄(Lysimachia christinae Hance)的新鲜或干燥全草，主产于四川、浙江等地。金钱草中主要药效成分为黄酮类化合物，据有关文献报导，其具有促进胆汁分泌和排泄、利尿排石等作用。本文主要研究内容和结果如下：
以金钱草总黄酮含量为指标，采用正交试验设计优选乙醇回流提取法提取金钱草总黄酮的最佳工艺条件为：80%乙醇20倍量，回流提取时间2.0h，提取次数为2次。与热水提取法相比，乙醇回流提取法具有更高的提取效率。
本实验取用D101型大孔树脂精制金钱草总黄酮，继而通过实验确定其最佳纯化工艺：上样浓度为0.2mg/mL，6倍量洗涤用水，吸附效果最优；以50%乙醇溶液洗脱，洗脱率达到最高。结果表明，其纯化效果优于醇沉法。
本文所建立的金钱草总黄酮提取、纯化工艺路线简单、高效，易于推广，为工业化提取和纯化金钱草总黄酮奠定了基础。

关键词  金钱草；总黄酮；提取；纯化；大孔树脂

Comparison of Extraction Technology of Total Flavonoids from Christina Loosestrife Herb
Abstract
Christina Loosestrife Herb has been long used as a Chinese traditional medicine，which is from flesh or dry plant of Lysimachia christinae Hance，family Primulaceae，This medicinal plant is mainly produced in Sichuan and Zhejiang Province. Flavonoids are the main active ingredients of Christina Loosestrife Herb. It is reported that the total flavonoids in this plant could promote biliation, egestion, urinate and remove urinary calculus. The main contents and conclusions are provided as follows:
The contents of total flavonoids from Christina Loosestrife Herb being served as index, in this topic, ethanol refluxing extraction were selected by orthogonal test. The results were shown that the optimal condition for procedure was extracting 2 h, 4 times with 20 folds of 80% ethanol. Compared with traditional hot water extraction, ethanol refluxing extraction has higher extraction efficiency.
 D101 macro porous resin was used , and the optimal purification conditions was obtained with 0.2mg/mL of total flavonoids concentration、6 folds of water and 50% ethanol as eluting agent. The results were shown that the purification effect was better than the method of ethanol precipitation.
Being simple, efficient and operable, the extraction and purification technology proposed in this research can be used to establish the foundation for extracting and purifying total flavonoids from Christina Loosestrife Herb on industrial scale.
Key words:  Christina Loosestrife Herb; total flavonoids; extraction; purification; macro porous resin
目  录
摘    要 I
Abstract II
第1章 绪  论 1
1.1 课题背景 1
1.2 金钱草的研究概况 1
1.2.1 金钱草中化学成分的研究概况 2
1.2.2 金钱草的药理作用 3
1.2.3 金钱草的临床应用 3
1.2.4 金钱草的不良反应 3
1.2.5 本节小结 4
1.3 黄酮类化合物的研究现状 4
1.3.1 黄酮类化合物的提取工艺研究 4
1.3.2 黄酮类化合物的纯化工艺研究 6
1.4 本课题研究的内容 7
第2章 金钱草中总黄酮提取工艺的比较研究 8
2.1 实验材料 8
2.1.1 仪器 8
2.1.2 药材与试剂 8
2.2 方法与结果 8
2.2.1 标准对照品溶液的制备 8
2.2.2 供试品溶液的制备 9
2.2.3 显色方法 9
2.2.4 芦丁标准品最大波长扫描 9
2.2.5 标准曲线的绘制 9
2.3 方法学考察 10
2.3.1 精密度试验 10
2.3.2 显色稳定性考察 10
2.3.3 重现性考察 11
2.3.4 加样回收率试验 11
2.4 乙醇回流提取金钱草中总黄酮 11
2.4.1 乙醇回流提取方法的单因素考察 11
2.4.2 正交试验 14
2.4.3 正交试验表的选择 14
2.5 热水回流提取金钱草中总黄酮 15
2.6 浸膏得率的测定 15
2.7 两种提取方法的提取效果比较 15
2.8 讨论 15
2.9 本章小结 16
第3章 金钱草中总黄酮纯化工艺的比较研究 17
3.1 实验材料 17
3.1.1 实验仪器 17
3.1.2 药材与试剂 17
3.2 D101型大孔树脂分离纯化金钱草总黄酮 17
3.2.1 大孔树脂的预处理及含水率的测定 17
3.2.2 大孔树脂的装柱 18
3.2.3 总黄酮粗提液的制备 18
3.2.4 树脂的静态吸附与解吸 18
3.2.5 D101型大孔吸附树脂分离纯化金钱草总黄酮工艺考察 18
3.2.6 D101型大孔吸附树脂吸附-洗脱过程动力学初探 20
3.2.7 大孔树脂的再生 21
3.3 传统有机溶剂纯化 21
3.4 两种纯化方法的纯化效果比较 21
3.5 讨论 22
3.6 本章小结 22
结  论 23
1 金钱草中总黄酮的提取 23
2 金钱草中总黄酮的纯化 23
致  谢 24
参考文献 25
附录 A 27
附录 B 32