Simultaneous determination of quercetin, kaempferol and isorhamnetin accumulated human beast cancer cells, by high-performance liquid chromatography

摘要

Quercetin, kaempferol and isorhamnetin aie the most important constituents in ginkgo fiavonoids. A simple, rapid and sensitive high-performance liquid chromatography method was developed to simultaneously determine quercetin, kaempferol and isorhamnetin absorped by human breast cancel cells. Cells were treated with ginkgo flavonols and then lysed with Triton-X 100. The flavonols in the samples were measured by RP-HPLC with a Cl 8 column after a simple extraction with a mixture of ether and acetone. The mobile phase contained phosphate buffer (pH 2,0; 10 mM) tetrahydrofuran, methanol and isopropanol (65:15:10:20, v/v/v/v). The ultraviolet detector was operated at 380nm. The calibration curve was linear from 0.1 to 1.0muM (r>0.999)for each flavonol. The mean extraction efficiency was about 70%, The recovery of the assay was between 98.9 and 100,6% The limit of detection was 0 01 (xM for quercetin and kaempferol and 0.05 muM for isorhamnetin. The limit of quantitation was 0.1muM (R.S.D. < 10%) for each flavonol. The intra- and inter-day coefficients of variation were less than 10% (R.S.D.). The validated method was applied to quantify quercetin, kaempferol and isorhamnetin in human breast cancer Bcap37 and Bcap37/MDR1 cells.