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目的:建立HPLC-DAD波长切换技术同时测定青钱柳提取物中异槲皮苷、槲皮素、山奈酚、山奈素、熊果酸5种有效成分的含量测定方法。方法:采用Thermo Syncronis C18色谱柱(250 mm×4.6 mm,5μm);流动相为乙腈-0.2%磷酸水溶液,流速1.0 m L·min-1,柱温35℃,进样量20μL,异槲皮苷、槲皮素、山奈酚、山奈素检测波长为360 nm,熊果酸检测波长为210 nm。结果:醇提物中各成分含量为0.6485%、0.0467%、0.0148%、0.0017%、0.0359%,水提物中异槲皮苷含量为0.0952%。异槲皮苷、槲皮素、山奈酚、山奈素、熊果酸的质量浓度分别在15.625~625μg·m L-1(r=0.9996)、4.05~162μg·m L-1(r=0.9998)、0.66~66μg·m L-1(r=0.9999)、0.09~28.8μg·m L-1(r=0.9998)、2.87~114.8μg·m L-1(r=0.9998)内与色谱峰峰面积呈良好的线性关系,平均回收率(n=6)均在100.24%、99.07%、101.52%、102.11%、97.24%(n=6)。结论:75%乙醇提取青钱柳中成分,基本可提取完全,残渣再用水提后只有微量的异槲皮苷;文章所建立的同时测定青钱柳中5种成分的含量,该方法简便、准确,具有较好的重复性和稳定性,为青钱柳质量的全面评价提供了科学依据。
OBJECTIVE: To establish a method for simultaneous determination of 5 active constituents of isoquercitrin, quercetin, kaempferol, kaempferol and ursolic acid by HPLC-DAD wavelength-switching technique. Methods: Thermo Syncronis C18 column (250 mm × 4.6 mm, 5 μm) was used. The mobile phase was acetonitrile-0.2% phosphoric acid solution and the flow rate was 1.0 m L · min-1. The column temperature was 35 ℃ and the injection volume was 20 μL. Glycosides, quercetin, kaempferol, kaempferol detection wavelength of 360 nm, ursolic acid detection wavelength of 210 nm. Results: The content of each component in the ethanol extract was 0.6485%, 0.0467%, 0.0148%, 0.0017% and 0.0359%, and the isoquercitrin content in the water extract was 0.0952%. The concentrations of isoquercitrin, quercetin, kaempferol, kaempferol and ursolic acid were in the ranges of 15.625 ~ 625μg · m L -1 (r = 0.9996) and 4.05 ~ 162μg · m L -1 (r = 0.9998) , 0.66 ~ 66μg · m L -1 (r = 0.9999), 0.09 ~ 28.8μg · m L -1 (r = 0.9998), 2.87 ~ 114.8μg · m L -1 (r = 0.9998) The average recovery was 100.24%, 99.07%, 101.52%, 102.11% and 97.24%, respectively (n = 6). Conclusion: 75% ethanol extract of Cymbidium officinale can be extracted completely, and only trace amounts of isoquercitrin can be extracted after the residue is extracted with water. The method of simultaneous determination of five components in Cyclocarya paliurus, Accurate, with good repeatability and stability, provide a scientific basis for a comprehensive evaluation of the quality of Cyclocarya paliurus.