RP-HPLC测定龙须藤中4种黄酮类成分的含量

RP-HPLC Determination of Four Flavonols in Bauhinia championii Benth.

关键词:  反相高效液相色谱法  龙须藤  杨梅素  槲皮素  表儿茶素  5,6,7,3',4',5'-六甲氧基黄酮

Key words:  RP-HPLC  Bauhinia championii Benth.  myricetin  quercetin  (-)-epicatechin  5,6,7,3',4',5'-hexamethoxy flavone

摘要:

目的用HPLC梯度洗脱法测定龙须藤中4种黄酮类成分的含量，考察了杨梅素、槲皮素、表儿茶素和5，6，7，3'，4'，5'-六甲氧基黄酮4种成分在不同产地药材中的含量。方法采用RP-HPLC测定，以Shimadzu C18 (4.6 mm′ 250 mm，5 μm)为色谱柱；流动相为乙腈-0.05%磷酸溶液，梯度洗脱(0 min，乙腈-0.05%磷酸比例为20∶80，5 min比例为30∶70；10 min，比例为50∶50；30 min，比例为50∶50)；流速1.0 mL·min-1，检测波长370 nm。结果杨梅素在0.071 6～0.716 μg、槲皮素在0.122 8～1.228 μg、表儿茶素在0.102 4～1.024μg、5,6,7,3',4',5'-六甲氧基黄酮在0.090 2～0.902 μg内呈良好的线性关系；重复性的RSD分别为1.32％，0.93％，1.02％，1.17％。不同产地中黄酮成分含量大小依次为福建青云山地区、广西隆安、江西安福、湖北宜昌、浙江缙云地区样品。结论本方法精密度、重复性和分离效果好，分析快速，适合于该药材的含量测定，不同产地中成分存在一定的差异。

Abstract:

OBJECTIVE To determine myricetin, quercetin, (-)-epicatechin, 5,6,7,3',4',5'-hexamethoxy flavone in Bauhinia championii Benth., several samples from different areas were examined, by HPLC using gradint elution. METHODS The determination was performed on a Shimadzu C18 column (4.6 mm′250 mm, 5 μm). The mobile phase consisted of acetonitrile-water, gradint elution (0 min, CH3CN-water 20∶80, 5 min, CH3CN-water 30∶70, 10 min, CH3CN-water 50∶50). The flow rate was 1.0 mL·min-1. The detection wavelength was set at 370 nm. RESULTS The line response range from 0.0716-0.716 μg of myricetin; 0.122 8-1.228 μg of quercetin; 0.102 4-1.024 μg of (-)-epicatechin; 0.090 2 -0.902 μg of 5,6,7,3',4',5'-hexamethoxy flavone. Their contents were highest in Qungyunshan, Fujian. CONCLUSION The RP-HPLC method shows a good separation，reproducity and accuracy. It will be used for determining flavones in herbs and extracts．The contents of four flavones are different in Bauhinia championii Benth. From different places.