title: [Determination of quercetin metabolism in UGT1A3 cDNA-expressing cells by RP-HPLC].
abstract: To develop a RP-HPLC method for the determination of quercetin in UGT1A3 cDNA-transfected cells.
  
  
  
  The lysate of cells transfected with human recombinant uridine 5-diphosphate glucuronosyltransferases UGT1A3 cDNA was co-incubated with quercetin, the reaction was terminated with acetonitrile, and luteolin was used as internal standard. The determination was performed on a C(1) reversed phase column with a mobile phase of methanol-0.1% formic acid (V/V) at a flow rate of 1.0 ml/min. The gradient elution was as follows: 0 - 25 min (30:70-80:20, methanol:0.1% formic acid), > 25-25.5 min (80:20), >25.5-27 min (80:20-30:70), > 27-30 min (30:70). A UV-VIS detector was operated at 368 nm.
  
  
  
  The standard curve was linear over the concentration range of 5-200 μmol/L (r = 0.9999). The limit of detection was 1.25 μmol/L(S/N ≥ 3), and the limit of quantification was 5 μmol/L (S/N >10, RSD = 6.99%). The method afforded recoveries of 99.1%-103.5%, and precisions for inter- and intra-assay were < 2.5% and < 8%, respectively. In addition, kinetic analysis indicated that the K(m), V(max) and CL(int) (V(max)/K(m)) values for quercetin glucuronide were (62.95 ± 13.16) μ mol/L, (284.50 ± 24.35)nmol*min⁻¹*g⁻¹ and 4.52 ml*min⁻¹*g⁻¹, respectively.
  
  
  
  The method established is accurate and simple and suitable for the determination of quercetin in UGT1A3 cDNA-expressed cells.
authors.affiliation: Department of Pharmaceutical Analysis and Drug Metabolism, College of Pharmaceutical Sciences, Zhejiang University, Hangzhou 310058, China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Yao
authors.forename: Xia
authors.initials: X
authors.lastname: Zhang
authors.forename: Yao
authors.initials: Y
authors.lastname: Liu
authors.forename: Lu-shan
authors.initials: LS
authors.lastname: Yu
authors.forename: Hui-di
authors.initials: HD
authors.lastname: Jiang
authors.forename: Su
authors.initials: S
authors.lastname: Zeng
mesh_terms: D002478:Cells, Cultured; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D014453:Glucuronosyltransferase / Q000235:genetics*; D006801:Humans; D011794:Quercetin / Q000032:analysis* / Q000493:pharmacokinetics; D014162:Transfection
chemical_list: D011794:Quercetin; C122075:UDP-glucuronosyltransferase, UGT1A3; D014453:Glucuronosyltransferase