丽江产玛咖指纹图谱分析及其活性组分提取分离研究
文献类型：学位论文
| 作者 | 王丽卫
|
| 学位类别 | 博士 |
| 答辩日期 | 2012-05-15 |
| 授予单位 | 中国科学院研究生院 |
| 导师 | 赵兵 |
| 关键词 | 玛咖 皂苷 生物碱 多糖 抗氧化 化学指纹图谱 |
| 其他题名 | Fingerprint of Maca from Lijiang and Extraction and Separation of Its Active Components |
| 学位专业 | 生物化工 |
| 中文摘要 | 玛咖（Lepidium meyenii）为十字花科独行菜属两年生植物，原产于秘鲁海拔3500m以上的安第斯山区，是一种药食两用植物。我国从2002年开始在中国丽江等地引种驯化，2011年被国家卫生部批准为新资源食品，目前云南丽江已成为全球最大的玛咖规模化种植基地，本论文针对丽江玛咖种植和加工过程中质量控制的难题和需求，围绕丽江产玛咖的功效物质成分开展了系统的研究，主要研究结果如下：（1）研究了丽江产玛咖不同形态类型不同生长时期的活性组分的累积规律。建立了同时检测玛咖中三种B族维生素的RP-HPLC方法，VB1、VB2和VB6的含量分别为748-2679mg/kg、10-145mg/kg和41-253mg/kg，是文献报道的几倍到几十倍；皂苷含量的分光光度计法表明：随着生长时期的增加，皂苷含量逐渐降低，9月份采集的玛咖中皂苷含量最高；总生物碱的酸性染料比色法表明：不同采收时期的玛咖中生物碱含量有显著差异，11月份采收的玛咖生物碱含量最高，有利于实现对种植过程的监控，提高玛咖种植产量和质量。（2）利用TLC方法对玛咖中的皂苷元和生物碱的种类进行了研究，发现不同玛咖形态类型的皂苷元和生物碱种类不同，但均含有一种相同的皂苷元和生物碱，生物碱的种类随着生长周期的增加而增多，12月份的玛咖形态类型中生物碱的种类相对较多。上述研究结果可用于监控玛咖种植过程、指导玛咖采收和提高玛咖质量。（3）以鲜玛咖根为原料进行了丽江产玛咖多糖的提取工艺优化，在提取温度80°C，料液比1:20，提取时间30min时，玛咖多糖提取率和多糖纯度的实际测定值为5.84%和88.78%，与理论值基本吻合，为玛咖功能成分的提取分离及玛咖功能性新产品研发生产提供了依据。（4）对不同玛咖形态类型的多糖分子量、单糖组成、红外谱图和抗氧化活性的研究结果表明：玛咖各形态类型多糖的分子量均小于100KDa；玛咖多糖主要由葡萄糖组成，其余单糖还有半乳糖、鼠李糖、阿拉伯糖、岩藻糖；不同形态类型的玛咖多糖均对超氧自由基具有明显的清除效果；且不同玛咖形态类型的多糖具有相同的红外谱图。（5）进行了丽江产玛咖不同极性提取物的抗氧化活性和抑菌活性的研究，结果表明：玛咖不同极性的提取物均有较高的还原活性及对超氧自由基和DPPH自由基的清除作用；玛咖抗氧化活性与总酚含量成正相关，且正丁醇提取物的抗氧化活性最强；除水提物外，其他5种不同极性提取物均有一定的抑菌效果，为阐明其构效关系和不同新产品开发提供理论依据。（6）对玛咖水提物和醇提物中的生物碱分别进行了分离纯化研究。确定了水提物中生物碱分离的填料、洗脱条件及最大上样量；首次利用薄层色谱纯化的方法从醇提物中分离得到了一种分子量为 268的咪唑类生物碱，为玛咖质量检测及控制奠定了基础。（7）利用内标法，建立了丽江产玛咖的化学指纹图谱，结果表明：玛咖各形态类型间的相似度均在0.88以上，所建立方法的精密性、稳定性和重复性良好，可用于玛咖种植、开发利用过程中质量实时监测和评价。 |
| 英文摘要 | Lepidium meyenii, usually named as maca, is a biennial plant belonging to the family of Brassicaceae. It is a medicinal and edible plant, which is mainly domesticated at altitudes above 3500m of the Peruvian Andes. It has been introduced to Lijiang since 2002, and was approved as a new resource of food by ministry of health of the people’s republic of Chinain 2011. Lijiang, has become the largest scale planting base for maca in the world. Focusing on the problem and need in the cultivation and processing quality control, a systematic study was carried out on the active components of Lijiang maca, and the major findings are as follows: （1）The accumulative rules of active components in different maca accessions under different growth periods were studied firstly. An HPLC method for simultaneous analysis of three kinds of vitamin B in maca was established, and the content of VB1, VB2 and VB6 were 748-2679mg/kg, 10-145mg/kg and 41-253mg/kg, respectively, which was significantly higher than those reported. A spectrophotometer method for the determination of saponin was established, and the result showed that saponin content decreased gradually with lengthening the growth period, and the highest saponin content was detected in September. Acid dyes colorimetric method was used to determinate maca alkaloid, and there was a significant difference in the alkaloid content of different growth periods. In November, alkaloid content reached the highest. These results could be beneficial to monitoring the cultivation process and improving the yield and quality of planting maca. （2）TLC method was used to study the types of sapogenin and alkaloid in maca. It was found that different maca accessions had different kinds of sapogenin and alkaloid. However, there was a common kind of sapogenin and alkaloid in different maca accessions. The type of alkaloids increased with lengthening the growth period, and was relatively higher in December. These results can be used to monitor the process of cultivation, guide harvest and improve the quality of maca. （3）Fresh maca sample was used to extract polysaccharides, and the optimal extraction conditions were as follows: extraction temperature was 80°C, solid-liquid ratio was 1:20 and extraction time was 30 min. The extraction rate and purity of polysaccharide were 5.84% and 88.78%, respectively, which was in agreement with the theoretical value. These results could provide basis for the large-scale production of maca functional component and functional product. （4）The polysaccharides from different maca accessions were examined for their molecular weights, monosaccharides compositions, infrared spectra and antioxidant activities. The results showed that polysaccharide molecular weights of different maca accessions were less than 100KDa, and were mainly composed of glucose and other monosaccharides, such as galactose, rhamnose, arabinose and fucose sugar. Different maca polysaccharides showed good scavenging effects on superoxide radicals, and they also had the same IR spectra. （5）Antioxidant activity and antibacterial activity of different polarity extracts obtained from Lijiang maca were studied. Results showed that all the extracts had a high reducing power and good radical-scavenging activity on superoxide anion radicals and DPPH radicals, and the antioxidant activity had a positive correlation with total phenol content. The n-butanol extract showed the most potent antioxidant activity. Other five polarity extracts had some antibacterial activity except for the aqueous extract. Clarification of relationship between the active components and activities could provide a theoretical basis for developing different types of new products. （6）Alkaloid separation from aqueous extract and alcohol extract of Lijiang maca was studied respectively. In the former separation, macroporous resin, elution condition and the maximum sample volume were studied. In the latter separation, a purification method by TLC was established, and finally an imidazole alkaloid with a molecular weight of 268 was isolated. These results could lay foundation for the quality control of maca. （7）An internal standard method was established in the chemical fingerprint analysis of the Lijiang maca. The method was validated to achieve the satisfactory precision, reproducibility and stability, and the similarities of different maca accessions were more than 0.88. These results could be used for the real-time monitoring and evaluation in the process of maca cultivation and development. |
| 语种 | 中文 |
| 公开日期 | 2013-09-25 |
| 源URL | [http://ir.ipe.ac.cn/handle/122111/1866]  |
| 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 | 王丽卫. 丽江产玛咖指纹图谱分析及其活性组分提取分离研究[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
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