高效液相色谱法测定玉米幼苗叶片中4种酚酸类化合物

Determination of Four Phenolic Acids in Leaves of Maize Seedlings by High Performance Liquid Chromatography

为研究干旱胁迫前后玉米幼苗叶片中4种酚类物质的变化,采用高效液相色谱法(HPLC),以Thermo Scientific Syncronis C18 (4.6 mm×250 mm,5μm)为分析柱,乙腈-0.5%乙酸溶液梯度洗脱,流速1.0 mL/min,检测波长280 nm,柱温30℃,对不同品种玉米幼苗叶片中的没食子酸、香豆酸、绿原酸、咖啡酸含量进行测定。结果表明,4种酚酸化合物的浓度在1～100 mg/L范围内与峰面积线性关系良好,相关系数R2均大于0.999,加标回收率和精密度RSD结果均满足分析要求;玉米幼苗叶片中香豆酸含量较高、没食子酸含量较少,干旱胁迫后除咖啡酸外其他3种酚酸含量均有下降;不同品种玉米幼苗叶片中4种酚类物质的含量不同,且干旱胁迫后酚酸的含量变化不同,与其对干旱的敏感程度有关。

The aim is to study the changes of four phenolic acids in the leaves of maize seedlings before and after drought stress.The content of gallic acid,coumalic acid,chlorogenic acid and caffeic acid were quantitatively determined by High Performance Liquid Chromatography(HPLC),Thermo Scientific Syncronis C18 column(4.6 mm×250 mm,5μm)as analytical column,acetonitrile-0.5%acetic acid solution with gradient elution,1.0 mL/min flow rate,280 nm detection wavelength and 30℃column temperature.The results showed that the linear relationship between peak area and 1-100 mg/L gallic acid,coumalic acid,chlorogenic acid and caffeic acid was good,the correlation coefficients(R2)were greater than 0.999.Adding standard recovery and precision RSD fulfilled the analysis requirements.The coumalic acid content was relatively high and gallic acid content was relatively low.After drought stress,the contents of the other three phenolic acids decreased except caffeic acid.The contents of four phenolic compounds in different varieties of maize seedlings was different and changed differently under drought stress,which was related to its sensitivity to drought.