Simultaneous determination of protocatechuic acid,chlorogenic acid and caffeic acid in Herba Patriniae by dual-wavelength HPLC

Objective: To develop a dual-wavelength HPLC method for the determination of protocatechuic acid,chlorogenic acid and caffeic acid contents in Herba Patriniae.Methods:The Odyssil C18 column(250 mm×4.6 mm,5 μm)was used with a mobile phase of acetonitrile-0.1% phosphate acid at 1.0 mL·min-1,and the detection wavelengths were set at 260 nm and 327 nm.The column temperature was controlled at 25 ℃ and the injection volume was 5 μL.Results:The method displayed good linearity over the concentration ranges of 3.42×10-2-3.42×10-1 μg(r =0.9997)for protocatechuic acid,4.66×10-2-4.66×10-1 μg(r =0.9999)for chlorogenic acid and 1.77×10-2-2.21×10-1 μg(r =0.9996)for caffeic acid,respectively.The average recoveries of protocatechuic acid,chlorogenic acid and caffeic acid were 101.6%,99.2% and 98.6%,with the RSD of 1.9%,1.7%,2.3%,respectively.Conclusion:The results indicated that the present RP-HPLC method is simple,accurate and reproducible,which is suitable for the quality control of protocatechuic acid,chlorogenic acid and caffeic acid in Herba Patriniae.