芦笋皂苷的纯化及活性研究
文献类型：学位论文
| 作者 | 赵芳春 |
| 学位类别 | 硕士 |
| 答辩日期 | 2012-05-01 |
| 授予单位 | 中国科学院研究生院 |
| 导师 | 赵兵 |
| 关键词 | 芦笋 甾体皂苷 大孔树脂 抑菌 抗氧化 口服液 |
| 其他题名 | Study on the Purification and Activity of Asparagus officinalis L. Saponins |
| 学位专业 | 生物化工 |
| 中文摘要 | 芦笋(Asparagus officinalis Linn)是一种药食两用的植物，药理学研究表明：长期食用芦笋可增强免疫力、延缓衰老、降血脂、降血糖、抗疲劳、抗肿瘤，其中甾体皂苷是其生理活性的重要物质基础。芦笋皂苷是一类具有环戊烷多氢菲母核结构的物质，有多种生物活性，高效的提取纯化工艺可为开发芦笋皂苷类药物和保健品奠定良好的基础。随着芦笋的大规模种植与产品上市，加工过程中产生了大量的边角料，大部分边角料被直接丢弃，即污染了环境，又浪费了资源。据测定，这些边角料中除含有丰富的营养物质外，还含有丰富的皂苷类成分，因此，亟需开发利用这些资源。课题组前期优化了芦笋废弃物中皂苷的提取工艺，但粗提物中皂苷纯度不高，在此基础上，本论文又开展了以下工作： (1) 建立了芦笋总皂苷含量测定方法，采用香草醛-高氯酸-冰醋酸体系做显色剂，在535nm下测定，在0~400μg范围内呈良好的线性关系。该方法简便，准确，适于芦笋总皂苷的含量检测。 (2) 对芦笋皂苷粗提物进行了主要成分分析，为纯化方案的确定奠定了基础。 (3) 通过泡沫实验、Libermannn-Burchard反应、Resen-Heimer 反应、Ehrish试剂反应确定了芦笋皂苷的主要类型：泡沫实验为阳性，证明有芦笋皂苷存在；Libermannn-Burchard反应最终颜色为绿色，表明存在甾体皂苷；Resen-Heimer反应最终呈现紫色，表明有甾体皂苷元存在；Ehrish试剂反应溶液呈红色，表明其甾体皂苷中存在呋喃型。 (4) 采用静态和动态吸附解吸试验优选适宜芦笋皂苷纯化的树脂并优化其工艺。通过静态吸附-解吸试验，对8种大孔吸附树脂纯化芦笋皂苷的效果进行比较，结果表明HPD-100树脂纯化芦笋皂苷效果最佳，并对HPD-100大孔树脂纯化芦笋皂苷工艺进行了优化。在上样质量浓度15mg/mL、溶液处理量150mL，1mL/min流速过柱；吸附后，先用50mL蒸馏水洗去杂质，再用200mL 80%乙醇洗脱，总皂苷的含量由14.26%提高到35.63%。将初次纯化产物按照上述工艺再纯化一次，总皂苷含量可高于50%，达到中药五类新药的含量要求。HPD-100可较好地吸附分离芦笋皂苷，操作简单、安全、成本低廉，有较高的应用价值。 (5) 考察了芦笋皂苷的体外抗氧化、抑菌活性。芦笋皂苷抗氧化能力随着浓度增大而增强，当增加到一定浓度后趋于稳定；相对而言，芦笋皂苷清除烷基自由基的能力最强，抑制DPPH自由基的能力次之，抑制超氧自由基的能力最弱。芦笋皂苷有较明显的抗氧化活性，可以考虑将其开发成为新型的抗氧化药剂。体外抑菌实验表明，芦笋皂苷对微生物有较好的抑制作用，抑菌活性：金黄色葡萄杆菌>大肠杆菌>枯草芽孢杆菌；且抑菌效果随皂苷浓度的升高而增强。 (6) 考察了芦笋皂苷口服液的稳定性，发现其对金属离子、温度、光照很敏感，因此该口服液应用棕色玻璃器皿盛装，在室温、避光下贮存。 |
| 英文摘要 | Asparagus officinalis Linn is an edible plant with curative effect. Studies on its pharmacology show that it has kinds of activities, such as improving immune, anti-aging, reducing blood lipid, lowering blood glucose, anti-fatigue and anti-tumor. The steroidal saponins in A. officinalis L. are important material basis. Asparagus saponins are a class of substances with cyclopentanoperhy drophenanthrene nucleus, and have various biological activities. Along with the enlargement of the planting areas year by year, a lot of leftover bits and pieces with no utilization are discarded, which pollute the environment and waste natural resources. It has been determined that these scraps are rich in nutrients and steroidal saponins. So, it is necessary to develop and utilize these resources. The technology for extracting total saponins from A. officinalis L. scraps were optimized by ultrasonic-assisted in our reseach group, basis on this work, this thesis mainly included the following aspects: (1) Determined the content of total saponins in A. officinalis L. by UV-spectral method. The result showed that the chromogenic reagent was vanillin-perchloric acid-glacial acetic acid and the detective wavelength was 535nm, the linear range was between 0~400μg.The method was simple, accurate and can be used as a quantitative analytical method for A. officinalis L. saponins. (2) Analyse the main components of the saponin in the crude extracts from A. officinalis L.. (3) The structure of A. officinalis L. saponins was investigated by poam reaction, Libermannn- Burchard reaction, Resen-Heimer reaction, Ehrish reaction. It was shown that the nucleus structure of A.officinalis L. saponins was steroidal saponins with furan-type. (4) Macro porous resin was chosen for the use of static and dynamic adsorption, desorption and purification experiments. Through static method, comparing the saponin purification result of 8 resins, HPD-100 resin was chosen due to its excellent adsorption and desorption performance. The optimal conditions were as follows: sample concentration 15mg/mL with loading amount of 150mL at a flow rate of 1mL/min, washing with 200ml 80% ethanol at a flow rate of 1mL/min. Sample was purified by HPD-100 resin according to these technology parameters and the content of saponins reached 35.63%. Then the primary purification product was purified in accordance with the above process, and the total content of saponins could be higher than 50% and complied with the standards of Class V of New Chinese Drug .These data demonstrated the excellence of HPD-100 resin in purifying saponins from A.officinalis L. and the advantages of the developed method for industrial application, like simple operation, safety, and low cost. (5) The pharmacological potentials of steroidal saponins from A.officinalis L.scraps were evaluated in vitro antioxidant and antimicrobial activities. Different scavenging activities on superoxide radical, DPPH radical, hydroxyl radical and alkyl radicals were measured. The inhibition of lipid peroxidation and reducing power were also observed. The results showed that the scavenging rate and reducing power increased with the increase of the concentration. In the case of antimicrobial activity, A. officinalis L. saponins possessed remarkable antimicrobial activities against 3 bacteria tested. The minimal inhibitory concentrations(MICs) of purified saponins against Staphylococci aureus, Escherichia coli, Bacillus subtilis were 30mg/mL、40mg/mL、50mg/mL, respectively. (6) The stability of A. officinalis L. saponin oral solution was investigated. It was suitable for storage in room temperature, dark and brown glassware. |
| 语种 | 中文 |
| 公开日期 | 2014-05-23 |
| 页码 | 93 |
| 源URL | [http://ir.ipe.ac.cn/handle/122111/8306]  |
| 专题 | 过程工程研究所_研究所（批量导入） |
| 推荐引用方式 GB/T 7714 | 赵芳春. 芦笋皂苷的纯化及活性研究[D]. 中国科学院研究生院. 2012. |
入库方式： OAI收割
来源：过程工程研究所
浏览0
下载0
收藏0
其他版本
除非特别说明，本系统中所有内容都受版权保护，并保留所有权利。