To establish a method for the content determination of 5-methylmellein, 5-hydroxymellein, 5-carboxylmellein and genistein in Wuling capsules simultaneously by HPLC. Four components were determined by HPLC on a Kromasil KR100-5C18 column (4.6 mm x 250 mm, 5 microm) with acetonitrile and 0. 2% phosphoric acid as mobile phase in a gradient elution. The flow rate was 1.0 mL x m min(-1), and the column temperature was set at 30 degrees C. 5-hydroxymellein showed a good linear relationship at the range of 7.86-157.2 ng, the average recovery was 101.0% with RSD 1.3%. 5-carboxylmellein showed a good linear relationship at the range of 9.57-191.4 ng, the average recovery was 98.6% with RSD 1.5%. Genistein showed a good linear relationship at the range of 28.80-576.0 ng, the average recovery was 98.6% with RSD 1.8%. 5-methylmellein showed a good linear relationship at the range of 21.46-429.2 ng, the average recovery was 99.2% with RSD 1.8%. The established method is feasible and the repeatability is good. The method can be used for quality control of Wuling capsules.