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.

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(4.6mm×250mm,5μm)为色谱柱;流动相为乙腈-0.05%磷酸溶液,梯度洗脱(0min,乙腈-0.05%磷酸比例为20∶80,5min比例为30∶70;10min,比例为50∶50;30min,比例为50∶50);流速1.0mL·min-1,检测波长370nm。结果杨梅素在0.0716～0.716μg、槲皮素在0.1228～1.228μg、表儿茶素在0.1024～1.024μg、5,6,7,3',4',5'-六甲氧基黄酮在0.0902～0.902μg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。

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 Ci8 column （4.6 mm×250 mm, 5 pro）. 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 pg 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.