OBJECTIVE：To establish a method for simultaneous determination of rutin， quercetin and nuciferine in Heye granule. METHODS：HPLC was performed on the column of Hypersil ODS2 with mobile phase of acetonitrile-water（containing 0.3％ phosphoric acid and 0.4％ triethylamine）（gradient elution） at a flow rate of 1.0 ml/min， the detection wavelength was 256 nm， the column temperature was 25 ℃， and the injection volume was 10 μl. RESULTS： The linear range was 0.012-0.240 μg for rutin（r＝0.999 9）， 0.010 4-0.208 μg for quercetin （r＝0.999 9） and 0.015-0.300 μg for nuciferine（r＝0.999 9）；RSDs of precision，stability and reproducibility tests were lower than 2.0％；recoveries were 98.5％-101.3％（RSD＝1.1％，n＝6），99.1％-101.6％（RSD＝1.0％，n＝6） and 98.9％-101.7％（RSD＝1.2％，n＝6）. CONCLUSIONS：The method is simple， accurate and reliable， and can provide reference for quality control of Heye granule.