Abstract:Objective: To establish an HPLC method for the simultaneous determination of quercetin and kaempferol in Ailanthi Fructus. Method: Quercetin and kaempferol were separated on Inertsil ODS-3 (4.6 mm×250 mm, 5 μm) column and detected at 360 nm. The mobile phase was methanol-0.4% phosphate (50:50). The flow rate was 1.0 mL·min-1. Result: Quercetin and kaempferol were linear within the range of 11.1-333 μg (r=1), 2.3-69 μg (r=1) respectively. The average recovery was 97.82% with RSD of 1.1%, 96.81% with RSD of 1.7% respectively. Conclusion: The method is simple, reproducible, and it can be used for the quality control of Ailanthi Fructus.