Rapid determination of six flavonoids from seabuckthorn leaves by RP-HPLC-DAD

To establish a rapid determination method of six flavonoids: catechin, rutin, myricetin, quercetin, kaempferol and isorhamnetin, from seabuckthorn leaves by RP-HPLC-DAD. The seabuckthorn leaves were first degreased by petroleum ether, extracted by ethanol, and determined by RP-HPLC-DAD. The six flavonoids were separated and eluted by a Shimadzu C₁₈(150 mm×2. 1 mm, 5 μm)column with methanol-water(0. 1% phosphoric acid)(60∶40)at a flow rate of 1. 0 mL/min. The detection wavelength were as follow: catechin 208 nm, rutin 257 nm, myricetin 373 nm, quercetin 371 nm, kaempferol 367 nm, and isorhamnetin 371 nm, respectively. The injection volume was 20 μL. The contents of the six flavonoids were in the range of 0. 47 to 30. 00 μg/mL with good linearity. The validation of the method, including precision, stability and recovery rate, was acceptable. The established method can be used for fast determination of the content of six flavonoids in seabuckthorn leaves.