款冬花欧盟质量标准研究

Research on Quality Standard of Farfarae Flos for EP Monograph

目的:根据《欧洲草药植物专论研究指导原则》建立款冬花药材的质量标准,主要包括薄层鉴别方法和含量测定方法。方法:薄层鉴别选用款冬酮作为指标成分,以环己烷-乙酸乙酯(4∶1)为展开剂,展开后喷以硫酸乙醇试液加热显色,置于紫外灯365 nm下检视;含量测定采用YMC-Pack ODS-A色谱柱,甲醇-水为流动相,梯度洗脱,柱温:35℃,流速:0.8 mL·min^(-1),检测波长:220 nm。结果:薄层鉴别方法专属性强,主成分条带清晰,辨识度高;含量测定方法学结果表明款冬酮在3.074~153.7μg·mL^(-1)内线性关系良好(r=0.999 9),平均回收率为99.9%(RSD=2.0%),重复性、中间精密度、稳定性及耐用性考察RSD均小于2.0%。结论:本研究中建立的薄层色谱鉴别方法和含量测定方法可准确测定款冬花中款冬酮的含量,为《欧洲药典》款冬花专论提供研究数据。

Objective： To establish the quality standard of Farfarae Flos mainly including TLC method and HPLC determination method, according to the technical guide for the elaboration of EP monographs. Methods： Tussilagone was selected as the marker constituent in the TLC identification method. After developing with cyclohexane-ethyl acetate（4：1）, the plate was sprayed with alcoholic solution of sulfuric acid Reagent and heated, and then examined under 365 nm UV light. In the HPLC determination method, a YMC-Pack ODS-A chromatographic column was used for the separation, with the mobile phase of methanol-water in a gradient elution procedure. The column temperature was set at 35 ℃, the flow rate was set as 0. 8 mL-min- and the wavelength was set at 220 nm. Results： The TLC identification method was established with good specificity, and the fluorescent zone of tussilagone was clearly detected with high distinguishing degree. Results： of HPLC method validation showedthat the determination method kept good linearity（ r = 0. 999 9 ）in the concentration range of 3. 074-153.7 μg·mL^-1 for tussilagone. The average recovery was 99.9% （ RSD = 2. 0% ） , RSDs of repeatability, intermediate precision, stability and robustness test were no more than 2. 0%. Conclusion： In this paper, a sensitive TLC identification method and an accurate HPLC determination method were established for the quality control of Faffarae Flos, which provided useful research data for EP monograph.