HPLC法测定芦笋茎秆中芦丁、槲皮素及白藜芦醇含量

HPLC Determination of Rutin, Quercetin and Resveratrol Content in Asparagus Stems

为了建立高效液相色谱法对芦笋茎秆中芦丁、槲皮素和白藜芦醇含量的测定方法。采用不同方法提取芦笋茎秆中的芦丁、槲皮素和白藜芦醇,得出甲醇法提取芦丁、甲醇-HCl法提取槲皮素以及乙醇法提取白藜芦醇得率最高;确定了高效液相色谱法对3种物质的最佳色谱条件为流动相:芦丁为0.2 mol/L乙酸钠-甲醇(Me-OH∶H2O=35∶65)溶液(用磷酸调p H 2.80)、槲皮素为无水甲醇-0.4%磷酸溶液(55∶45,V/V),等梯度洗脱,白藜芦醇为水和甲醇,二元线性梯度洗脱;检测波长分别为:254 nm、360 nm和306 nm;流速:0.8 m L/min、1.0 m L/min、0.8 m L/min。通过该方法测得芦丁、槲皮素及白藜芦醇的平均回收率分别为:98%、97%、98%。

In order to establish a HPLC method to determinate its content of rutin,quercetin and resveratrol. The study used different methods to extract rutin,quercetin and resveratrol from asparagus stems,the results showed that methanol,methanolHCl and ethanol had the best extraction rate for rutin, quercetin and resveratrol extraction. And it chose the optimum chromatographic conditions of HPLC method： The mobile phases of rutin and quercetin were 0.2 mol / L sodium acetate-methanol（ Me-OH ∶H2O = 35∶65） solution（ p H 2.80 adjusted with phosphoric acid）,anhydrous methanol-0.4% phosphoric acid solution（ 55∶ 45,V/V）,isocratic elution,and resveratrol mobile phase was water and methanol,binary linear gradient elution,respectively. Their detection wavelength was 254 nm,360 nm and 306 nm,and velocity of flow was 0.8 m L / min,1.0 m L / min and 0.8 m L / min. The average recoveries of rutin,quercetin and resveratrol were 98%,97% and 98%.