Rapid determination of 13 phenolic acids in barley grain by reversed phase high performance liquid chromatography

A reversed phase high performance liquid chromatographic method for the rapid determination of 13 phenolic acids(including gallic acid,protocatechuic acid,chlorogenic acid,p-hydroxybenzoic acid,caffeic acid,vanillic acid,syringic acid,m-hydroxybenzoic acid,p-coumaric acid,ferulic acid,veratric acid,o-coumaric acid,and salicylic acid) in barley grain was developed.The separation was performed on a SUPELCO Ascentis C18 column(150 mm×4.6 mm,5 μm) at 40℃ by gradient elution.The mobile phase consisted of methanol and 0.1% formic acid solution at a flow rate of 0.7 mL/min.The injection volume was 10 μL,and the detection wavelength was 275nm.All of the phenolic acids were well separated within 20 min.There were good linear correlations between the concentrations and peak areas of the 13 components with the correlation coefficients all above 0.999.The detection limits of the components were 0.30~4.99 mg/L,and the average recoveries were 87.09%~105.5% with relative standard deviations of 0.04%~3.1%.Good repetition and precision of the method were demonstrated through the results,and it can be applied to the rapid determination and evaluation of phenolic acids in barley grain.