HPLC-DAD法测定铁筷子中香草酸和阿魏酸的含量

Determination of Vanillic Acid and Ferulic Acid in Iron Chopstick by HPLC-DAD Method

本研究旨在建立高效液相色谱法(HPLC)-二极管阵列检测器(DAD)测定铁筷子药材中香草酸和阿魏酸的含量。采用ZORBAX Eclipse Plus C18色谱柱(4.6mm×25.0 mm,5μm);流动相为乙腈(A)-0.2%磷酸水(B),梯度洗脱,流速1.0 mL/min;柱温25℃;检测波长(0~15 min,280 nm,香草酸;15~70 min,327 nm,阿魏酸);进样量10μL。香草酸、阿魏酸的分离度均>1.5,分离效果好,分别在22.72~727.00μg/mL、9.69~310.00μg/mL范围内呈良好线性关系,r> 0.999;平均回收率(RSD)分别为101.08%(3.09%)、98.98%(3.11%),铁筷子样品S1中阿魏酸和香草酸的含量最高,S1中香草酸含量为(0.77±1.42) mg/g,阿魏酸含量为(0.36±1.32)mg/g。本研究建立的HPLC-DAD波长切换法灵敏、准确,重复性好,可用于铁筷子药材中香草酸和阿魏酸含量的测定。

A high performance liquid chromatography(HPLC)-Diode Array Detector(DAD)method was established for the determination of vanillic acid and ferulic acid in iron chopsticks herbs.The chromatography was performed on ZORBAXEclipse Plus C18 column(4.6 mm×250 mm,5μm).The mobile phase was acetonitrile(A)-0.2%phosphoric acid water(B),gradient elution,flow rate 1.0 mL/min;column temperature 25℃;detection wavelength(0~15 min,280 nm,vanillic acid;15-70 min,327 nm,ferulic acid);the injection volume was 10 L.The separation degree of vanillic acid and ferulic acid was more than 1.5,and the separation effect was good,and the linear relationship was good in the range of 22.72~727.00 g/mL and 9.69~310.00 g/mL,respectively,with r>0.999.The average recoveries(RSD)were 101.08%(3.09%)and 98.98%(3.11%),respectively.The contents of ferulic acid and vanillic acid in iron chopsticks herbs S1 were the highest,the vanillic acid content in S1 was(0.77±1.42)mg/g and ferulic acid content was(0.36±1.32)mg/g.This study established HPLCDADswitching wavelength method is sensitive,accurate and reproducible,and can be used for the determination of vanillic acid and ferulic acid in the iron chopsticks herbs.