A method for the determination of xanthohumol in beer using solid-phase extraction-high performance liquid chromatography (SPE-HPLC) has been developed. A Waters Sep-Pak C18 column was used to extract and clean-up the sample. The separation was achieved on a reversed-phase Agilent Zorbax Eclipse XDB-C18 (250 mm x 4.6 mm, 5 microm) in a linear gradient, and the mobile phases were consisted of water (containing 0.1% formic acid) (A) and methanol (B) with a flow rate of 0.4 mL/min. In addition, the column temperature was maintained at 25 degrees C. The detection wavelength was set at 370 nm. There was a good linear relationship (r2 = 1) in the range of 0.5 - 500 microg/L. The average recoveries were between 91.21% and 95.58% with the relative standard deviations less than 2%. The limit of detection was 0.24 microg/L and the limit of quantification was 0.80 microg/L. It was proved to be a convenient and accurate method for the analysis of xanthohumol content in beer.