Abstract: To develop a sensitive and specific reversed-phase high performance liquid chromatography (RP-HPLC) method for simultaneous determination of three flavoniods in Polygonum viviparum, the HPLC separation was performed on a Kromasil C18 column (250 mm×4.6 mm, 5 μm) and detected with DAD at 360nm. The gradient elution was carried out with methano-H2O (containing 0.25% phosphoric acid) as the mobile phase at a flow rate of 1 mL/min. The results indicated that the calibration curves for viterxin, quercetin and quercitrin were linear over the ranges of 8-40 μg/mL, 5-50 μg/mL and 5.33-52 μg/mL, their assay recoveries were 100.3%, 98.9% and 100.4%, their intraday repeatability were 0.78%, 0.51% and 0.38%, and interday repeatability were 2.6%, 1.9% and 2.1%, respectively. This method was simple, accurate and sensitive, so it could be used for the quality control of P. viviparum.

Key words: Polygonum viviparum, viterxin, quercetin, quercitrin, RP-HPLC