基于薄层色谱与一测多评法的积雪草提取物质量评价研究

Research on quality evaluation of Centella asiatica extract based on thin layer chromatography and quantitative analysis of multi-components by single marker

中文关键词:  积雪草  提取物  羟基积雪草苷  薄层色谱法  一测多评

英文关键词:Centella asiatica  extract  madecassoside  thin layer chromatography  quantitative analysis of multi-components by single marker

中文摘要:

      目的 建立积雪草提取物的薄层色谱法(thin-layer chromatography, TLC)鉴别方法和高效液相色谱定量方法, 并结合一测多评法(quantitative analysis of multi-components by single marker, QAMS)实现积雪草苷、积雪草苷B和羟基积雪草苷的定量测定。方法 首先采用TLC对积雪草提取物进行定性鉴别, 确保样品的真实性; 其次采用高效液相色谱法进行含量测定, 流动相分别为2 mmol/L β-环糊精溶液和乙腈, 检测波长为205 nm。以羟基积雪草苷为参照物, 计算其他两种物质的相对校正因子, 并将测定含量与外标法测定结果进行比较。结果 3种成分在10~500 mg/L范围内线性关系良好, 不同浓度水平的加标回收率范围为87.0%~108.2%, 相对标准偏差为3.1%~5.2%, 方法具有良好的精密度、重复性和稳定性。QAMS与外标法测定积雪草提取物中3种特征性成分的含量一致性良好。结论 TLC鉴别方法结合QAMS方法实现定性定量检测, 能够对积雪草提取物的质量控制提供技术支持。

英文摘要:

      Objective To establish a thin-layer chromatography (TLC) method for identification of Centella asiatica extracts, as well as a liquid chromatography method to achieve the quantitative determination of asiaticoside, asiaticoside B and madecassoside by means of quantitative analysis of multi-components by single marker (QAMS). Methods Firstly, TLC was used for the qualitative identification of Centella asiatica extracts to ensure the authenticity. Then, for content determination, high performance liquid chromatography was ultilized and the mobile phases were 2 mmol/L β-cyclodextrin solution and acetonitrile, respectively. The detection wavelength was set at 205 nm. Madecassoside was used as the reference substance to calculate the relative correction factors of other 2 components. Finally, the content determined by QAMS was compared with the results determined by the external standard method. Results The 3 kinds of ingredients showed good linear relationships in the range of 10–500 mg/L. The spiked recoveries at varied concentration levels ranged from 87.0% to 108.2%, with the relative standard deviation of 3.1%–5.2%. The method demonstrated excellent precision, repeatability and stability. The content of the 3 kinds of characteristic components in Centella asiatica extracts determined by QAMS and the external standard method showed good consistency. Conclusion The TLC method for identification coupled with the established QAMS method for quantification can provide technical support for the quality control of Centella asiatica extracts.