OBJECTIVE: To establish a method for simultaneous determination of paeoniflorin, naringin, neohesperidin, notoginsenoside R1, ginsenoside Rg1 and ginsenoside Rb1 in Quyu zhitong mixture. METHODS: HPLC method was adopted. The determination was performed on Waters Xbridge C18 column with mobile phase consisted of 0.1% phosphoric acid-acetonitrile (gradient elution) at the flow rate of 1.0 mL/min. The detection wavelength was set at 203 nm, and column temperature was 20 ℃. The sample size was 5 μL. RESULTS: The linear ranges of paeoniflorin, naringin, neohesperidin, notoginsenoside R1, ginsenoside Rg1 and ginsenoside Rb1 were 0.273 4-2.734 μg(r=0.999 9),0.119 1-1.191 μg(r=0.999 9),0.081 5-0.815 μg(r=0.999 9),0.622 8-6.228 μg(r=0.999 9),0.807 2-8.072 μg(r=0.999 9),1.036 4-10.364 μg(r=0.999 9). The limits of quantitation were 0.082 0, 0.029 8, 0.028 5, 0.436 0, 0.403 6, 0.310 9 μg. The limits of detection were 0.027 9, 0.009 5, 0.010 2, 0.124 6, 0.121 1,0.093 3 μg. RSDs of precision, stability and reproducibility tests were lower than 2.0%. Recoveries were 97.85%-100.34%(RSD=0.81%,n=6),98.14%-101.22%(RSD=1.09%,n=6),98.42%-102.15%(RSD=1.29%,n=6), 97.77%-100.25%(RSD=0.96%,n=6),97.32%-99.53%(RSD=0.81%,n=6) and 98.28%-101.51% (RSD=1.11%,n=6). CONCLUSIONS: The method is simple, rapid, stable, feasible and reproducible, and can be used for the simultaneous determination 6 components in Quyu zhitong mixture.