HPLC同时测定三叶青中5种黄酮类成分的含量

Simultaneous Determination of Five Major Flavonoids in Tetrastigma Hemsleyanum by HPLC

关键词:  三叶青  芦丁  异槲皮苷  山奈酚-3-O-芸香糖苷  槲皮素  山奈酚  含量测定

Key words:  Tetrastigma hemsleyanum  rutin  isoquercitrin  kaempferol-3-O-rutinoside  quercetin  kaempferol  determination

摘要:

目的 建立HPLC同时测定三叶青中5个黄酮类化合物芦丁、异槲皮苷、山奈酚-3-O-芸香糖苷、槲皮素和山奈酚的含量。方法 Agilent ZORBAX SB-C₁₈色谱柱（250 mm×4.6 mm，5 μm）；流动相为乙腈-0.2%磷酸溶液，梯度洗脱，流速为1.0 mL·min^-1，检测波长为360 nm，柱温为35℃。结果 芦丁、异槲皮苷、山奈酚-3-O-芸香糖苷、槲皮素和山奈酚分别在0.087 8~0.878 μg、0.069 6~0.696 μg、0.162 9~1.629 μg、0.002 7~0.027 μg、0.007 23~0.072 3 μg内与峰面积呈良好的线性关系，r ≥ 0.999 7；平均加样回收率分别为97.44%，96.88%，97.00%，96.42%，96.24%。结论 该方法简便、高效，可作为三叶青的质量控制提供参考。

Abstract:

OBJECTIVE To develop a determination method of rutin, isoquercitrin, kaempferol-3-O-rutinoside, quercetin, and kaempferol in Tetrastigma hemsleyanum by HPLC. METHODS HPLC assay was carried out using an Agilent ZORBAX SB-C₁₈ column (250 mm×4.6 mm, 5 μm)with column temperature at 35℃. The mobile phase was acetonitrile-0.2% phosphoric acid in gradient elution at a flow rate of 1.0 mL·min^-1. And the detection wavelength was 360 nm. RESULTS The linear ranges of rutin, isoquercitrin, kaempferol-3-O-rutinoside, quercetin, kaempferol, and baicalin were 0.087 8-0.878 μg, 0.069 6-0.696 μg, 0.162 9-1.629 μg, 0.002 7-0.027 μg, 0.007 23-0.072 3 μg, respectively (r ≥ 0.999 7), and the average recoveries of three components were 97.44%, 96.88%, 97.00%, 96.42%, 96.24%, respectively. CONCLUSION The method is simple, quick and accurate, can be used for the quality control of the Tetrastigma hemsleyanum.