白桦树中桦木醇的提取与桦木酸合成研究
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湖南省教育厅资助项目(08C433),湖南省研究生创新项目(14)


Study on extraction of betulin from the white birch and synthesis of betulinic acid
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    摘要:

    为大量获得高纯度的桦木酸及准确测定桦木酸的含量,从白桦树皮中提取桦木醇,经琼斯试剂氧化制备中间产物桦木酮酸后,用硼氢化钠还原合成桦木酸,采用IR、HPLC-MS、1H-NMR以及13C-NMR对桦木醇和桦木酸的结构进行鉴定,确定最佳提取与合成条件,并建立了HPLC测定桦木酸含量的方法:色谱柱Zorbax Eclipse XDB-C8(4.6 mm×150 mm,5 μm);流动相∶甲醇-水(4 g/L甲酸铵)=40∶60,流速1.0 mL/min, 波长210 nm,柱温为室温.结果表明,桦木醇最佳提取条件:甲醇为溶剂,固液比为15/200(g/mL),70 ℃,提取3 h;桦木酸最佳合成条件为:桦木醇与琼斯试剂的摩尔比为1∶6,20 ℃反应3 h.桦木酸的线性范围为0.031 25~0.5 mg/mL,回归方程为Y=379 381X-2 397(R2=0.998 8),平均回收率可达97.97%,桦木酸含量为 96.53%.

    Abstract:

    In order to get plenty of betulinic acid with high purity and test the content of betulinic acid,betulinic acid was synthesized by oxidizing betulin from the bark of the white birth with Jone’s reagent,followed by selective In order to get plenty of betulinic acid with high purity and test the content of betulinic acid, betulinic acid was synthesized by oxidizing betulin from the bark of the white birth with Jone’s reagent, followed by selective oxidation of the resulting betulonic acid with NaBH4. IR, HPLC-MS,1H-NMR and 13C-NMR were used to identify the structure of betulin and betulinic acid. To compare extraction efficiency, several solvents were evaluated, also extraction time and extraction time were optimized. A quantitative analysis method of betulinic acid by HPLC was formed with ethanol-water (4 g/L ammonium formate) as the mobile phase on Zorbax Eclipse XDB-C8 column (4.6 mm×150 mm, 5 μm).The flow-rate was 1.0 mL/min and the UV detection was set at 210 nm, with a column of room temperature.The results showed that the optimal extraction conditions were that the white birch bark was extracted 3 h with methanol under reflux at 70 ℃ and the ratio of the bark to the extraction solvent was 15/200(g/mL). The optimal synthesis conditions were that the ratio of betulin to Jone’s reagent was 1∶6 and the mixture was reacted 3 hours at 20 ℃. A good linearity of betulinic acid was in the range of 0.031 25 mg/mL to 0.5 mg/mL (Y=379 381X-2 397,R2=0.998 8). The recovery rate was 97.97%, the RSD (n=9) was 1.64%, and the content of betulinic acid was 96.53%.

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易金娥,文利新,袁莉芸,B. Obminska-Mrukowicz,袁慧.白桦树中桦木醇的提取与桦木酸合成研究[J].湖南农业大学学报:自然科学版,2010,36(5):574-580.

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  • 收稿日期:2009-12-11
  • 最后修改日期:2010-01-25
  • 录用日期:2010-09-29
  • 在线发布日期: 2010-10-16
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