Simultaneous quantitative determination of quercetin,kaempferol and isorhamnetin in Hippohae rhamnoids granules under tri-wave length by RP-HPLC

OBJECTIVE To assay the content of quercetin,kaempferol and isorhamnetin in Hippohae rhamnoids Linn.granules under tri-wave length by HPLC.METHODS Flavone glycosides were extracted in two steps from hippohae rhamnoids Linn.granules,in which the quercetin,kaempferol and isorhamnetin content were assayed.The RP-HPLC workstation equipped with the Symmetry C18 column(250 mm×4.6 mm,5.0 μm) at room temperature was used,the mobile phase was composed of acetotrile and 0.4% of phosphoric acid(4:6) with a flow rate of 1.0 mL·min-1.Quercetin was detected at 266 nm,while kaempferol at 360 nm and isorhamnetin at 370 nm.RESULTS The good linearity was obtained at the concentration of 0.2-1.6 μg,the average recoveries were 99.7%,98.3% and 101.0%,with the relative coefficients of 0.9991,0.9994 and 0.9990,respectively.CONCLUSION The method is proved to be sensitive and specific,applicable for the determination of quercetin,kaempferol and isorhamnetin in Hippohae rhamnoids Linn.granules.