Objective: A method was established by HPLC for the determination of four flavonoids in Nelumbinis Folium to supply further basis to the quality control of Nelumbinis Folium. Method: Chromatography conditions were Kromasil C18 (4.6 mm×200 mm

5 μm) column

methanol (A)-0.2% phosphric acid system as mobile phase

(0-8 min

10%-40%A;8-23 min

40%-70%A;23-30 min

70%-85%A);the flow rate was 1 mL·min-1 and column temperature was set at 30 ℃. Diode array detector was used and detected at 360 nm. Result: The assay displayed good linearity;the linear ranges of quercetin-3-O-sambubioside

hyperin

isoquercitrin and quercetin were 6.10-97.60

18.76-300.16

8.48-135.68

9.12-145.92 mg·L-1 respectively. The average recoveries were 100.98% (RSD 1.85%)

99.05% (RSD 1.48%)

99.29% (RSD 1.84%)

97.71% (RSD 1.78%) respectively. The content of the four flavonoids in Nelumbinis Folium from five different origins was 0.092%-0.161%

0.949%-1.862%

0.067%-0.133% and 0.040%-0.062%. Conclusion: The method is simple

accurate and can be used to determine quercetin-3-O-sambubioside

hyperin

isoquercitrin and quercetin in Nelumbinis Folium. The content of four flavonoids in Nelumbinis Folium from different origins varied notably and should be taken into the quality control of Nelumbinis Folium.