Simultaneous determination of quercetin, kaempferol and isorhamnetin accumulated human breast cancer cells, by high-performance liquid chromatography.
Quercetin, kaempferol and isorhamnetin are the most important constituents in ginkgo flavonoids. A simple, rapid and sensitive high-performance liquid chromatography method was developed to simultaneously determine quercetin, kaempferol and isorhamnetin absorped by human breast cancer 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 C18 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 380 nm. The calibration curve was linear from 0.1 to 1.0 microM (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 microM for quercetin and kaempferol and 0.05 microM for isorhamnetin. The limit of quantitation was 0.1 microM (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.
Wang
Yi
Y
Department of Pharmaceutical Analysis and Drug Metabolism, College of Pharmaceutical Sciences, Zhejiang University, Hangzhou, PR China.
Cao
Jiang
J
Weng
Jian-Hua
JH
Zeng
Su
S
D001943:Breast Neoplasms / Q000737:chemistry*; D002138:Calibration; D045744:Cell Line, Tumor; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D044948:Flavonols / Q000032:analysis*; D006801:Humans; D044949:Kaempferols / Q000032:analysis*; D000091402:Prohibitins; D011794:Quercetin / Q000032:analysis* / Q000031:analogs & derivatives; D012015:Reference Standards; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013056:Spectrophotometry, Ultraviolet
D044948:Flavonols; D044949:Kaempferols; C000717331:PHB2 protein, human; D000091402:Prohibitins; C047368:3-methylquercetin; C006552:kaempferol; D011794:Quercetin