Objective To establish a method for the determination of rutin and quercetin in Melastoma dodecandrum by HPLC. Methods The chromatographic conditions were as follows：with the SERVO PT-C18 column（250mm×4.6mm， 5μm） as chromatographic column， methanol（A） -0. 2% phosphoric acid（B） as mobile phase， gradient elution： 0～28min， 31% A； 28～28.1min， 31%A～44% A； 28. 1～45min， 44% A， 255nm as the detection wavelength， 35℃ as the column temperature， 1.0mL·min-1 as the flow rate. Results The good linear range for rutin was 10.35～82.80μg·mL-1（r＝0.9998） and for quercetin was 4.05～32.40μg·mL-1（r＝0. 9997）. The average recovery was 91. 09%（RSD＝0. 87%） for rutin and 93. 97%（RSD＝1. 00%） for quercetin. The content range of rutin and quercetin was 0. 857～0. 933mg·g-1， 0. 306 ～0. 333mg·g-1 in 3 batches of herbs， respectively. Conclusion The method is simple， accurate and repeatable， and it is suitable for content determination of rutin and quercetin in Melastoma dodecandrum.