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HPLC法测定经酸水解的菊花中三种活性黄酮苷元的含量

李福高, 周慧, 孙冬黎, 王如伟, 曾苏, 蒋惠娣*
  

  1. 1. 浙江大学 药学院 药物分析与代谢研究室, 浙江 杭州 310058
    2. 浙江康恩贝药业有限公司, 浙江 杭州 310051
    3. 杭州保灵养泰禾生物科技有限公司, 浙江 杭州 310022
  • 收稿日期:2008-10-17 修回日期:2009-02-10 出版日期:2009-03-15 发布日期:2009-03-15
  • 通讯作者: 蒋惠娣*

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*

摘要:

本文建立了一种测定菊花主要生物活性黄酮的方法。将粉碎后的菊花经盐酸甲醇混合溶液水解、提取, HPLC法测定其木犀草素、芹菜素、香叶木素活性成分的含量。采用正交设计法优化提取水解条件, 其最佳条件为: 粉末状菊花在盐酸浓度为2.4 M, 甲醇浓度为80% (v/v) 混合溶液中, 80 ºC水浴回流 2 h。提取液采用高效液相色谱法分析, SB-C18为分离柱, 检测波长为344 nm木犀草素、芹菜素、香叶木素的标准曲线范围分别为2.233–71.46 μg/mL, 1.668–53.38 μg/mL, 1.410–45.11 μg/mL; 回收率分别为100.3%, 103.9%, 100.4%; 日内精密度分别为0.68%, 0.21%, 0.38%; 日间精密度分别为2.6%, 1.7%, 2.0%。采用上述方法测定了杭白菊等8种主流菊花中上述3种活性苷元的含量。

关键词: 菊花, 木犀草素, 芹菜素, 香叶木素, 水解, 测定

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

中图分类号: 

Supporting: Foundation items: The Ministry of Sciences and Technology of China (Grant No. 2006BAI06A18-11), Administration and Traditional Chinese Medicine, Zhejiang, China (Grant No. 2007ZA012).
*Corresponding author. Tel./fax: 86-571-88208408