摘要
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 （HC1） 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 HC1 for 2 h in 80 ℃ 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.
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,用 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) 的混合溶液中, 于80 oC水浴回流 2 h。提取液采用高效液相色谱法分析,以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2.233-71.46μg/mL,1.668-53.38μg/mL,1.410-45.11μg/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。
基金
The Ministry of Sciences and Technology of China (Grant No. 2006BAI06A18-11)
Administration and Traditional Chinese Medicine,Zhejiang,China (Grant No. 2007ZA012)
