Objective: To establish an RP-HPLC method for simultaneous determination of contents of the four flavonoids

astilbin

neoisoastilbin

quercetin-3-O-α-L-rhamnoside

engeletin

contained in Smilax china. Method: Hypersil C18 (4.6 mm×250 mm

5 μm) column was used with the mobile phase of acetonitrile-water (0.04% phosphoric acid) at a flow rate of 0.7 mL·min-1 and the detection wavelength of 290 nm. External standard method (astilbin

neoisoastilbin

quercetin-3-O-α-L-rhamnoside

engeletin used as the reference substance) and quantitative analysis on multi-components by single marker method (astilbin used as the internal reference substance) were adopted to determinate the contents of the four flavones in S. china.Result: The four flavonoids showed good linear relationship (r>0.999 9) within the linear range of calibration curve by the external standard method. The average recoveries of astilbin

neoisoastilbin

quercetin-3-O-α-L-rhamnoside

engeletin were 100.8%

100.3%

101.4%

101.4%

respectively;and the RSD of the above four components were 1.8%

2%

1.4%

1.6%

respectively. The relative correlation factors (RCF) of neoisoastilbin

quercetin-3-O-α-L-rhamnoside

engeletin were 0.726

0.264

0.801

respectively and the RSD were 0.6%

0.7%

1.2%

1.1%

respectively by the quantitative analysis on multi-components by single marker (QAMS) method. No significant difference between the contents of QAMS method and external standard method was observed. Conclusion: The present research established an HPLC method proved to be easy and accurate to determine the contents of the four flavones in S. china. There is no significant difference between the contents of the four flavones in S. china. in the results of QAMS method and external standard method. The QAMS method could replace the external standard method to determine the contents of the four flavones in S. china and can be used for the quality control of S. china.