Simultaneous determination of quercetin,kaempferol and isorhamnetin in Hippophae rhamnoides L.fruit by HPLC

OBJECTIVE To develop an HPLC method for simultaneous determination of three flavonoids such as quercetin,kaempferol and isorhamnetin in Hippophae rhamnoides L.fruit.METHODS The column was Kromasil-C18(250 mm×4.6 mm,5 μm),the mobile phase was acetonitrile(A)-0.6% phosphoric acid solution(B) with linear gradient elution(0-15 min,33%→32% A;15-30 min,32%→31% A),the flow rate was 1.0 mL·min-1,the column temperature was 30 ℃ and the detection wavelength was 370 nm.RESULTS The method had good linear relationship within the range of 1.68-26.88 μg·mL-1 for quercetin,1.52-24.32 μg·mL-1 for kaempferol,and 1.76-28.16 μg·mL-1 for isorhamnetin.The average recoveries of quercetin,kaempferol and isorhamnetin were 98.55%,97.63% and 97.98%,respectively and the RSD were 1.48%,1.66% and 1.54%,respectively.CONCLUSION This method is simple,accurate and reliable.It is can be used to determine the contents of quercetin,kaempferol and isorhamnetin in Hippophae rhamnoides L.fruit.