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An efficient method for the simultaneous determination of three flavone aglycones in Flos Chrysanthemi by acid hydrolysis and HPLC

Fu-Gao Li, Hui Zhou, Do-Lin Sun, Ru-Wei Wang, Su Zeng, Hui-Di Jiang*

  1. 1. Department of Pharmaceutical Analysis and Drug Metabolism, College of Pharmaceutical Sciences,
    Zhejiang University, Hangzhou 310058, China
    2. Zhejiang Conba Pharmaceutical Co. Ltd. Hangzhou 310051, China
    3. Hangzhou Youither Bioscience Co. Ltd. Hangzhou 310022, China
  • Received:2008-10-17 Revised:2009-02-10 Online:2009-03-15 Published:2009-03-15
  • Contact: Hui-Di Jiang*

Abstract: An efficient method for the simultaneous quantification of three flavone aglycones was developed and validated for the control of flavonoids in Flos Chrysanthemi (FC). The method employed mixed solvent of hydrochloric acid (HCl) and methanol to extract and hydrolyze flavone glycosides simultaneously from powdered FC followed by HPLC to determine the contents of three flavone aglycones, luteolin, apigenin and diosmetin, which are the major bioactive flavones in FC. The extraction and hydrolysis conditions optimized by the orthogonal tests were as follows: powdered FC was refluxed in 80% methanol (v/v) containing 2.4 M HCl for 2 h in 80 ºC water bath. The HPLC separation was performed on C18 column and detected with DAD at 344 nm. The calibration curves for luteolin, apigenin and diosmetin were linear over the ranges of 2.233-71.46 μg/mL, 1.668-53.38 μg/mL and 1.410-45.11 μg/mL, their assay recoveries were 100.3%, 103.9% and 100.4%, their intra-day repeatability (R.S.D.) were 0.68%, 0.21% and 0.38% and their inter-day repeatability were 2.6%, 1.7% and 2.0%, respectively. The developed method has been successfully utilized to assay eight species of popular FC samples, especially to analyze Hangbaiju, the well-known FC in China.

Key words: Chrysanthemum morifolium Ramat., Chrysanthemum morifolium Ramat., Luteolin, Luteolin, Apigenin, Apigenin, Diosmetin, Diosmetin, Hydrolysis, Hydrolysis, Determination, Determination

CLC Number: