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Study on Separation and Purification of Polysaccharides from Gingko Biloba Sarcotesta and Their Biological Activities

Author: ZuoGuangHua
Tutor: WuXiangYang
School: Jiangsu University
Course: Of Food Science
Keywords: Ginkgo biloba exocarp Polysaccharides Phenol - sulfuric acid method Purification Acetylation Immunocompetent MTT assay In vitro antitumor activity
CLC: R284
Type: Master's thesis
Year: 2007
Downloads: 205
Quote: 1
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Abstract

Ginkgo is the world's precious herbs and medicinal plants, and its leaves, fruits and testa mused has medicinal value for development, known as the \According to statistics, China Ginkgo has been cultivated throughout the country 27 provinces (city), an annual output of more than 12,000 tons ginkgo ginkgo, outer seed coat portion representing approximately 70% of the entire ginkgo fruit, which estimated that China has at least 30,000 tons per year Fresh Ginkgo biloba exocarp above, if the removal of which 60% water, dried Ginkgo biloba exocarp more than 12,000 tons per year. Developed by the Ginkgo biloba extract from ginkgo (EGb) and preparations are now welcomed by the people of natural medicine by ginkgo developed with a variety of nutritious foods are also many, only Ginkgo biloba exocarp utilization rate low. Polysaccharide is an important bioactive components. Studies have reported water extract of Ginkgo biloba exocarp polysaccharides have anti-tumor, immunomodulatory and anti-aging and other active, therefore, research and development of agricultural waste Ginkgo biloba exocarp, you can change waste into treasure, ginkgo resource utilization. In this thesis, Ginkgo biloba exocarp as raw material, the Ginkgo biloba exocarp polysaccharides extraction and determination methods, Ginkgo biloba exocarp polysaccharides purification techniques, removal of protein polysaccharides and polysaccharide immune activity and pure polysaccharide and its acetylated The antitumor activity of the product, the main research contents and results are as follows: 1. Studied the water extraction extract of Ginkgo biloba exocarp polysaccharides conditions. With Ginkgo biloba exocarp polysaccharides was investigated indicators, on the basis of single factor experiment, using three factors and three levels Orthogonal the extraction process, get the best extraction conditions were: temperature 100 ℃, solid-liquid ratio 1:12, time 1h and extract 3 times, under the process conditions Extraction rate 95.09%; three factors on Ginkgo biloba exocarp polysaccharides extraction rate of the order of extraction temperature> Time> solid-liquid ratio. 2. Studied the phenol - sulfuric acid method of Ginkgo biloba exocarp polysaccharides approach. On the basis of single factor experiments, using three factors and three levels orthogonal measurement conditions were optimized to obtain the best measurement conditions as follows: 5% phenol solution 1.2 mL, amount of sulfuric acid 4mL, color temperature and color rendering time 30 ℃ 20min , amount of sulfuric acid is an important factor affecting the color. The method of recovery was 99.60%, RSD = 3.02% (n = 5). 3. Study of Ginkgo biloba exocarp polysaccharides (GBEP-1) and de-proteoglycan (GBEP-2) immune activity. The results show that GBEP-1 low-dose group, GBEP-2 low-dose group phagocytic percentage phagocytic index with the model group were no significant differences in the GBEP-1 high-dose group, GBEP-2 high-dose group phagocytic percentage phagocytic index with the model group were significantly different (p <0.05 or p <0.01). GBEP-1, GBEP-2 administration group phagocytic percentage of phagocytic index has further enhanced the trend, and has a certain dose-effect relationships; GBEP-1, GBEP-2 low-dose group and the model group level hemolysin no significant differences, and GBEP-1, GBEP-2 high-dose group hemolysin level with the model group had a greater significant difference (p <0.001). As the dose increases, GBEP-1, GBEP-2 administration group serum hemolysin antibody levels have a tendency to increase further, and has a certain dose-effect relationships; GBEP-1 low-dose group, GBEP-1 dose Groups and GBEP-2 low-dose group for thymus index and the model group had no significant difference, GBEP-1 high-dose group, GBEP-1 high-dose group, GBEP-2 low-dose group, GBEP-2 high-dose group thymus, spleen index with the model group were significantly different (p <0.05 or p <0.01). 4. Using DEAE-52 cellulose and Sephadex G-100 column, right Ginkgo biloba exocarp polysaccharides purified polysaccharide obtained four GBEPP11, GBEPP22, GBEPP33 and GBEPP44, purity of 90.87%, 95.36 %, 92.57% and 94.63%; while using high performance liquid chromatography (HPLC) and gas chromatography (GS) method for the determination of the molecular weight and monosaccharide composition, molecular weight fraction obtained four 950604,311288,47987 and 3373, respectively , monosaccharide composition of four components are: GBEPP11 composed of rhamnose, glucose and galactose, the molar ratio of 16:10:0.96; GBEPP22 composed of rhamnose, glucose and galactose, the molar ratio of 2.64:1:1.43; GBEPP33 composed of rhamnose, glucose and galactose, the molar ratio of 37.01:1:8.46; GBEPP44 composed of rhamnose and glucose in a molar ratio of 1.9:1. 5. MTT assay was used to study the Ginkgo biloba exocarp polysaccharides, removal of protein polysaccharides and polysaccharide components and GBEPP11 pure acetylated product of human gastric cancer cell line (AGS), human gastric cancer cell line (SGC), human leukemia cell lines (U937 ) inhibitory effect in vitro results show that these three tumor cells they have different degrees of inhibition. Which GBEPP11 acetylated product antitumor activity better than the other samples.

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