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| 粗毛纤孔菌总三萜的纯化及对胆酸盐结合能力分析 |
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刘爽爽1,2,刘东超1,2,孙凡越1,2,李德海1,2*,王占斌1,2
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| (1.东北林业大学 林学院, 哈尔滨 150040;2.黑龙江省森林食品资源利用重点实验室, 哈尔滨 150040) |
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| 摘要: |
| 为提高粗毛纤孔菌菌丝体和子实体总三萜的纯度及体外胆酸盐结合能力，研究有机溶剂、高径比(硅胶高∶硅胶柱直径)、上样量和硅胶量质量比、洗脱体系和洗脱剂比例对总三萜纯度的影响。并且在最佳的纯化条件下与其它纯化方法作对比。通过红外光谱对总三萜的结构进行初步分析。并通过体外模拟体内环境试验对比了粗提物和纯化物总三萜的体外胆酸盐结合能力。结果表明:在高径比55∶22、上样量和硅胶量质量比4∶80、洗脱体系为丙酮∶石油醚、洗脱剂体积比3∶1时，粗毛纤孔菌菌丝体和子实体乙酸乙酯萃取后的总三萜纯度最高分别为65.35%、42.32%，是粗提物总三萜的4.26和4.60 倍，该技术与大孔树脂和单纯硅胶纯化技术相比菌丝体与子实体总三萜纯度均最大。红外光谱分析显示，菌丝体和子实体总三萜存在—CH2、—CH3、—S-O等官能团的差异，但都符合三萜类物质的特征结构。粗毛纤孔菌菌丝体和子实体总三萜纯化后体外胆酸盐结合能力明显提高，其中菌丝体总三萜的胆酸钠、甘氨胆酸钠和牛磺胆酸钠结合能力最强，分别为1.14、1.78和2.06 μmol/100 mg，是菌丝体粗提物总三萜胆酸盐结合能力的1.56、1.73和1.72 倍。 |
| 关键词: 粗毛纤孔菌 总三萜 纯化 萃取 胆酸盐 |
| DOI：10.11841/j.issn.1007-4333.2020.05.11 |
| 投稿时间：2019-10-15 |
| 基金项目:中央高校基本科研业务费专项资金资助(2572019BA04)；哈尔滨市科技局创新研究基金项目(2017RAQXJ091) |
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| Purification of total triterpenoids from Inonotus hispidus and analysis on bile salt binding ability |
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LIU Shuangshuang1,2,LIU Dongchao1,2,SUN Fanyue1,2,LI Dehai1,2*,WANG Zhanbin1,2
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| (1.College of Forestry, Northeast Forest University, Harbin 150040, China;2.Key Laboratory of Forest Food Resources Utilization of Heilongjiang Province, Harbin 150040, China) |
| Abstract: |
| This study aims to improve the purity of total triterpenoids of mycelium and fruiting body from Inonotus hispidus. The effect of organic solvent, height-to-diameter ratio(height of silica gel∶diameter of silica gel), sample mass-to-silica gel mass, and eluent ratio on purity of total triterpenoids were studied. Then, it is compared with other purification methods. The structure of total triterpenoids was analyzed by infrared spectroscopy. The bile salt binding abilities of the crude extract and the purified material were determined by in vitro simulation. The results showed that the optimal parameters were height-to-diameter ratio of 55∶22, sample mass-to-silica gel mass of 4∶80, acetone and petroleum ether as elution system with ratio of 3∶1. Under this condition, the highest purity of total triterpenoids extracted with ethyl acetate from mycelium and fruiting body were 65. 35% and 42. 32%, which were 4. 26 and 4. 60 times of total triterpenoids from the crude extract, respectively. This method attained the highest purity comparing with other results purified using macroporous resin and silica gel. Infrared spectroscopy analysis showed that there were differences between total triterpenoids from mycelium and fruiting body in —CH2, —CH3, —S-O, and etc, but they both conformed to the characteristic structure of triterpenoids. The bile salt binding ability of total triterpenoids was improved significantly after purification. The binding ability of total triterpenoids from mycelium with sodium cholate, sodium glycocholate and sodium taurocholate were the strongest. The highest values were 1. 14, 1. 78 and 2. 06 μmol/100 mg, which were respectively 1. 56, 1. 73 and 1. 72 times of crude extract from mycelium. |
| Key words: Inonotus hispidus triterpenoids purification extraction bile salt |
