Simultaneous determination of chlorogenic acid,puerarin and paeoniflorin in Kangganning Mixture by multiple wavelength RP-HPLC

AIM: To establish a method for simultaneously determining chlorogenic acid,puerarin and paeoniflorin in Kangganning Mixture(Flos Lonicerae japonicae,Radix Puerariae Lobatae,Radix Paeoniale rubra,Fructus Forsythiae,etc.). METHODS: A multiple wavelength HPLC method was devoloped.The analysis was performed on an Agilent Zorbax SB C_(18) column(250 mm×4.6 mm,5 μm) with acetonitrile-0.1%H_3PO_4(11∶89) as the mobile phase.The detection wavelengthes were monitored at 324 nm,254 nm and 230 nm for chlorogenic acid,puerarin and paeoniflorin,respectively.The flow rate was 1.0 mL/min and the column temperature was at 30 ℃.(RESULTS:) The calibration curves of chlorogenic acid,puerarin and paeoniflorin showed good linearity at the ranges of 0.179-2.864 μg,0.071 55-1.144 8 μg and 0.372 5-5.96 μg,respectively.The average recoveries were(100.7%,) 103.3%,102.6% with RSD of 2.2%,2.4%,1.9%,respectively. CONCLUSION: The method is simple,accurate and reproducible for determining chlorogenic acid,puerarin and paeoniflorin in Kangganning Mixture.