【作者】 白秀君；

【导师】 杨海燕； 蒋益虹；

【作者基本信息】 新疆农业大学 ， 食品科学， 2007， 硕士

【摘要】 荷叶为卫生部批准的“可用于保健食品的物品”,现代临床广泛将荷叶应用于肥胖症及高血脂症,并取得了较好的疗效。药理研究显示荷叶生物碱类具有明显降脂减肥、抑制胆固醇、抗有丝分裂、抑菌和止痉挛等药理活性。目前有关荷叶生物碱的分离和制备的研究较少,本文主要报道了荷叶生物碱的提取工艺的优化、分离纯化进行了一系列的研究,为荷叶生物碱的进一步深入研究奠定了基础。现将主要研究结果报告如下:一荷叶生物碱的提取工艺采用热水提取法从荷叶中提取生物碱,对影响荷叶生物碱提取的四个主要因素――提取时间、提取温度、pH、固液比,在进行单因素实验的基础上,设计了L₁₆（4⁴）正交实验,以生物碱的提取率为指标,并通过极差、方差分析对这些因素进行了统计分析,并用人工神经网络对生物碱的提取工艺进行优化。确立了荷叶生物碱热水提取工艺:pH值2.5,提取温度85℃,提取时间1.75h,固液比1:35。二荷叶生物碱薄层层析鉴别荷叶生物碱在硅胶薄层板上用溶剂展开体系三氯甲烷:甲醇:二乙胺（90:9:1,V/V）展开,使荷叶中生物碱达到了较好的分离效果,此条件也可用于检测分离纯化效果。三高速逆流色谱法（HSCCC）分离纯化荷叶中的生物碱首次采用HSCCC成功分离了荷叶中的生物碱化合物。选择三氯甲烷:甲醇:pH4.00的水（4:3:2,v/v）为溶剂系统进行分离纯化,得到了三种组分,含量分别为94.13%、89.68%、92.25%。四高效液相色谱法（HPLC）分离纯化及定量分析建立了荷叶碱的高效液相色谱法（HPLC）定量分析,并利用制备型高效液相色谱对荷叶生物碱进行单体分离,分别得到了含量为98.78%、97.63%、99.99%的三种单体生物碱化合物。研究表明,采用人工神经网络和传统的正交实验相结合的分析方法建立了荷叶生物碱的提取工艺。薄层层析法对荷叶碱粗提物进行鉴别,高速逆流色谱和高效液相色谱两种不同方法都成功制备荷叶生物碱单体。更多还原

【Abstract】 A traditional Chinese herb called lotus leaf is approved as‘material that can be used in healthy foods’by Ministry of Health, P. R. China.In the present study, there were few paper about the separation and preparation of lotus leaf alkaloids. In this paper, the technology for extraction of alkaloid from lotus leaf was studied. Quantitative analysis of nuciferine by HPLC was established here. Two methods-high speed countercurrent chromatography and high performance liquid chromatography for separation and purification of alkaloids were applied. The main results were described as follows:1. The technology for extraction of alkaloidThe technology for extraction of alkaloid from lotus leaf was studied. Through combination of artificial neural network (ANN) with traditional orthogonal design, a new method for analyzing, processing and optimizing the test data was put forward. The optimum condition for extraction was summarized, namely, pH value 2.5,extracting temp. 85℃, duration of extraction 1.75h and solid to liquid ratio 1:35. The satisfactory results confirmed the reasonableness and practicability.2. Alkaloids of lotus leaf were identified by TLCUsing thin layer chromatography(TLC) for identifying the alkaloid in the extract demonstrated that the kinds of alkaloid changed with the extract solvent. Chloroform-methanol- lignocaine amine (90:9:1)mixture as a mobile phase. The nuciferine in the lotus leaf were found.3. Separation and purification of alkaloids from Lotus Leaf by high speed countercurrent chromatographyHigh speed countercurrent chromatography (HSCCC) was applied to the separation of alkaloids from Lotus Leaf the first time. High speed countercurrent chromatography with a two-phase solvent system composed of Chloroform:methanol:pH4.0 water(4:3:2,volume ratio). Five collections were separated. The results were verified by TLC, in which three components exhibited orange-red colour with bismuth potassium iodide reagent. HSCCC is a convenient, feasible and effective technique for the separation and purification of active components in Chinese herbal medicines.4. Separation and purification alkaloids from Lotus Leaf by high performance liquid chromatography. And found the method of HPLC analytical of nuciferine.A method using preparative high performance liquid chromatography (HPLC) to isolate four individual alkaloids from lotus leaf was investigated. A preparative C18 column was employed in the procedure. First, by isocratic elution with methanol:water:lignocaine amine (75:25:0.0125,V/V) as mobile phase at a flow rate of 8 ml/min, two mixture (part1, part2) were obtained. Then, by isocratic elution with acetonitrile:water:lignocaine amin(e55:45:0.0125,60:60:0.0125)as mobile phase at a flow rate of 8 ml/min for part1 and part2, respectively, three substance were obtained. HPLC analytical results showed that the purity of each alkaloid was over 97%. Under the chromatographic condition, every peak was completely baseline separated.In a word, the technology for extraction of alkaloid from lotus leaf was founded. High speed countercurrent chromatography (HSCCC) and performance liquid chromatography (HPLC) are two efficient methods for separation and preparation of alkaloid.更多还原