积雪草苷提取纯化工艺研究

论文编号:ZY074    论文字数:10700,页数:28

摘 要：本文首先建立了HPLC法测定积雪草苷提取物中积雪草苷含量的方法，采用Diamonsil C18柱，乙腈-水（50:50，v/v）为流动相，检测波长 200nm，结果积雪草苷加标回收率为 100.42℅，RSD为 1.44℅。此方法简单、准确，适用于积雪草苷提取物的质量控制。然后对积雪草有效成分积雪草苷的提取纯化工艺进行了研究。以积雪草苷含量为评价指标，采用 L9(33)正交试验优选积雪草苷提取工艺，考察了乙醇浓度、料液比、提取时间对积雪草苷含量的影响，最佳工艺为 60℅ 乙醇料液比 1：15回流提取 2h。减压回收乙醇，得积雪草苷醇提部分。加适量水溶解，以石油醚-乙酸乙酯-正丁醇溶剂系统萃取，正丁醇部分减压蒸干，溶于少量甲醇，然后滴加丙酮，析出积雪草苷粗品；并与另一条路线：氯仿-正丁醇溶剂系统萃取，正丁醇部分减压蒸干得积雪草苷粗品进行回收率和含量的比较，得到较优提纯工艺。最后采用薄层色谱法对积雪草苷粗品进行检测，摸索不同展开剂及其配比对积雪草苷粗品的分离情况，以氯仿-甲醇-水（60: 15: 10）及正丁醇-乙酸乙酯-水（4：1：5）的分离效果最好，可为后续进一步的纯化提供依据。
关键词：积雪草苷；提取；分离；纯化

Abstract: First of all, in this essay established a HPLC method asiaticoside extract content asiaticoside method.Using Diamonsil C18 reserved-phase column, mobile phase as acetonitrile-water （50:50，v/v）elution, detect wavelength at 200nm. The average recovery of asiaticoside is100.42 %, and RSD is 1.44%. This method is simple, accurate and applicable to extract asiaticoside quality control. And then the active ingredients of Centella asiatica asiaticoside the extraction and purification process were studied. Asiaticoside in content for evaluation,To use of L9 (33) orthogonal test asiaticoside preferred extraction process to inspect the ethanol concentration, liquid ratio, extraction time asiaticoside content, the best technology for liquid ethanol 60 ℅ return than 1:15 Extraction of  2h. Decompression ethanol recovery may asiaticoside ethanolic part. Add appropriate amount of water dissolved in petroleum ether - ethyl acetate - n-butanol solvent extraction systems, dry vacuum parts n-butanol, dissolved in a small amount of methanol, and then dropwise acetone, precipitated crude asiaticoside; and another route: chloroform - n-butanol solvent extraction system, vacuum dry n-butanol part of a crude asiaticoside to compare the recovery rate and the content, the better purification process. Finally, the use of thin-layer chromatography of crude asiaticoside test, carried out to explore the different agents and their ratio on asiaticoside separation of crude to chloroform - methanol - water (60: 15: 10) and is butanol - ethyl acetate - water (4:1:5) the best separation for the follow-up to provide a basis for further purification.
Keywords：Asiaticoside; extraction; separation; purification

目  录
中文摘要 II
英文摘要 III
目录 IV
1. 绪论 1
 1.1 积雪草苷的理化性质与结构 1
 1.2 积雪草苷的药理作用 2
 1.3 积雪草苷提取纯化工艺的研究 3
 1.4 积雪草苷分析方法的研究 4
2. 实验部分 5
 2.1 实验材料和仪器 5
 2.2 积雪草苷的 HPLC方法建立 6
 2.3 正交试验提取工艺设计 10
 2.4 积雪草苷的分离纯化 12
 2.5 薄层色谱法鉴别积雪草苷粗品 14
3. 结果与讨论 16
 3.1 积雪草苷提取工艺的结果与讨论 16
 3.2 积雪草苷纯化工艺路线的结果与讨论 19
 3.3 薄层色谱展开剂优化实验结果与讨论 20
4．总结与展望 23
致谢 24
参考文献 25