Determination of Kaempferol and Quercetin in Smilax china by RP-HPLC

OBJECTIVE To establish a RP-HPLC method for the determination of kaempferol and quercetin in Smilax china L.,and to investigate their contents in different places.METHODS The determination was performed on a Lichrospher C_(18) column(4.6 mm×250 mm,5 μm).The detection wavelength was at 365 nm,with the mobile phase of Water-Acetonitrile-Phosphoric-Triethylamine(65∶35∶0.27∶0.45) at the flow rate of 0.7 mL·min~(-1).RESULTS Kaempferol,quercetin and impurities were separated.The theoretical plate numbers of kaempferol and quercetin were 5 121 and 6 673,the resolution between kaempferol,quercetin and impurities were 1.96 and 5.32,respectively.All the parameters were suitable for determination.Both of kaempferol and quercetin showed a good linearity over the range of 5～100 mg·L~(-1)(r=0.999 4,0.999 6),respectively,the average recoveries of kaempferol and quercetin were 105.67% and 87.58%,respectively.Their contents were highest in Zhang jia jie,Hunan and lowest in Chibi,Hubei.CONCLUSION The RP-HPLC method shows a good separation,reproducity and accuracy.It will be used for determining kaempferol and quercetin in herbs and extracts.The contents of kaempferol and quercetin are different in Smilax china L.from different places.