中国中药杂志

2009, v.34(04) 423-427

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HPLC测定萹蓄中槲皮素和山柰素的含量
Determination of quercetin and kaempferol in Polygonum aviculare by HPLC

陈娟;师彦平;
CHEN Juan,SHI Yanping(Key Laboratory for Natural Medicine of Gansu Province,Lanzhou Institute of Chemical Physics,Chinese Academy of Sciences,Lanzhou 730000,China)

摘要(Abstract):

目的:建立同时测定中药萹蓄中槲皮素和山柰素含量的高效液相色谱方法。方法:采用KromasilC18色谱柱(4.6mm×250mm,5μm),以甲醇-0.1%磷酸溶液(65:35)为流动相,流速1.0mL·min-1,检测波长为366nm。对样品提取和水解过程中的各个影响因素进行了考察。结果:甲醇是萹蓄中槲皮素和山柰素的最佳提取溶剂,65%甲醇是槲皮素和山柰素及其苷的最佳提取溶剂,优化的水解条件为采用4.0mol·L-1盐酸溶液,80℃水浴上水解1.0h。槲皮素和山柰素分别在0.58~362,0.51~320mg·L-1峰面积与浓度呈良好的线性关系(r=0.9999),最低检出限分别为0.03,0.02mg·L-1;平均加样回收率分别为99.7%和99.0%,RSD分别为1.7%和3.9%。结论:该方法简便、准确、重现性好,可用于中药萹蓄的质量控制。
Objective:To develop a high performance liquid chromatographic method for the simultaneous determination of quercetin and kaempferol in Polygonum aviculare.Method:The optimized method was achieved for the separation and detection of quercetin and kaempferol using Kromasil C18 column(4.6 mm×250 mm,5 μm) as the stationary phase,methanol-0.1% aqueous phosphoric acid solution(65:35) as the mobile phase at a flow rate of 1.0 mL·min-1,366 nm as the detection wavelength.The main factors,extraction solvent extraction time,hydrolysis tine,temperature and hydrochloric acid concentration,which influenced the sample extraction and hydrolysis procedure,were intensively explored.Result:Methanol and 65% methanol were chosen as the extracting solvent for the free compounds and for the total compounds,respectively.The optimized hydrolysis procedure was that the sample was hydrolyzed 1.0 h with 4.0 mol·L-1 hydrochloric acid at 80 ℃.Quercetin and kaempferol showed good relationship at the range of 0.58-362 mg·L-1 and 0.51-320 mg·L-1,respectively.Both of the correlation coefficients of the calibration curves were 0.999 9.The detection limits of quercetin and kaempferol were 0.03 mg·L-1 and 0.02 mg·L-1 at a signal-to-noise ratio of 3,respectively.The average recoveries of quercetin and kaempferol were 99.7% and 99.0% with relative standard derivation(RSD) of 1.7% and 3.9%.Conclusion:The results showed that the method is simple,accurate and repeatable and it is suitable for quality control of Polygonum aviculare.

关键词(KeyWords): 高效液相色谱;萹蓄;槲皮素;山柰素
high performance liquid chromatography;Polygonum aviculare;quercetin;kaempferol

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基金项目(Foundation): 国家科技支撑计划项目(2007AA09Z403);; 中国科学院科技支撑项目

作者(Author): 陈娟;师彦平;
CHEN Juan,SHI Yanping(Key Laboratory for Natural Medicine of Gansu Province,Lanzhou Institute of Chemical Physics,Chinese Academy of Sciences,Lanzhou 730000,China)

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