三叶青地上部分抗炎提取物质量控制研究

Studies on Quality Control of Anti-inflammatory Extracts of Tetrastigma Hemsleyanum Diels Et Gilg's Aerial Part

关键词:  三叶青地上部分提取物  质量控制  薄层色谱法  比色法  高效液相色谱法  一测多评

Key words:  extracts of Tetrastigma hemsleyanum Diels et Gilg’s aerial part  quality control  TLC  colorimetry  HPLC  quantitative analysis of multi-components by single-marker (QAMS)

摘要:

目的 建立三叶青地上部分抗炎提取物（anti-inflammatory extracts of Tetrastigma hemsleyanum Diels et Gilg’s aerial part，THAA）质量控制方法。方法 采用薄层色谱法鉴别THAA；采用比色法测定THAA总黄酮含量；建立HPLC同时测定THAA 7个效应组分含量，并在此基础上建立了以异荭草苷为内参物，同步测定7个效应组分的HPLC一测多评法。结果 6批THAA供试品色谱，在与三叶青地上部分对照药材及混合对照品色谱相应位置上，显相同颜色的荧光斑点，各组分分离良好；建立的比色法在9.63~57.79 μg·mL^-1内呈良好的线性关系，回归方程为A=1.120×10^-2C-2.400×10^-3，r=0.999 5，3批THAA总黄酮平均含量为60.02%~60.32%。HPLC测得THAA中绿原酸、荭草苷、异荭草苷、异荭草苷-2’-O-鼠李糖苷、牡荆苷、异牡荆苷、牡荆苷-2’’-O-鼠李糖苷的平均含量分别为21.64%，8.04%，6.61%，19.51%，2.03%，1.59%，4.03%；一测多评法同时测定THAA中7个抗炎效应组分含量与外标法分别测定的含量结果无显著性差异。结论 所建立的薄层色谱鉴别法，总黄酮的含量测定法以及7个效应组分HPLC含量测定法及一测多评法，可为THAA内在质量控制提供参考。

Abstract:

OBJECTIVE To develop a method for studies on the quality control of anti-inflammatory extracts of Tetrastigma hemsleyanum Diels et Gilg's aerial part(THAA). METHODS Identify the THAA by TLC; the content of total flavonoids of THAA wene determined by colorimetric method; HPLC was established for the simultaneous determination of 7 pharmacodynamic components of THAA. Moreover, isoorientin was used as the internal reference in the determination of 7 pharmacodynamic components by quantitative analysis of multi-components by single-marker for THAA. RESULTS Six batches of THAA samples were chromatographed, and the fluorescent spots of the same color were displayed on the corresponding positions of the medicinal materials and the mixed reference materials on THAA. The components were well separated. The established colorimetric method showed a good linear relationship in the range of 9.63-57.79 μg×mL^-1. The regression equation was A=1.120×10^-2 C-2.400×10^-3, r=0.999 5. The average content of total flavonoids in 3 batches of THAA was 60.02%-60.32%. The content of chlorogenic acid, orientin, isoorientin, isoorientin-2'-O-rhamnoside, vitexin, isovitexin, vitexin-2''-O-rhamnoside in the anti-inflammatory extracts of THAA was about 21.64%, 8.04%, 6.61%, 19.51%, 2.03%, 1.59%, 4.03%; There was no significant difference between the content of 7 anti-inflammatory medicinal components in the THAA quantitative analysis of multi-components by single-marker multi-evaluation method and HPLC. CONCLUSION The established TLC identification, the determination method of total flavonoids, and the HPLC and quantitative analysis of multi-components by single-marker of the 7 pharmacodynamic components can be used as a reference for the internal quality control of the THAA.