Determination of 3',4',5',5,7-pentamethoxyflavone in the plasma and intestinal mucosa of mice by HPLC with UV detection.
In a preliminary experiment 3',4',5',5,7-pentamethoxyflavone (PMF) inhibited adenoma development in Apc(Min) mice, a model of the human heritable condition familial adenomatous polyposis. An HPLC method for tricin was modified and validated to permit measurement of PMF in mouse plasma and intestinal mucosa. HPLC analysis was carried out on a Hypersil-BDS C(18) column with detection at 324 nm and tricin as internal standard. The assay was linear in the range of 100-2000 ng/mL plasma and 1.0-40 microg/mL mucosa. PMF in plasma was efficiently extracted using solid-phase columns. In the case of mucosa organic solvent protein precipitation displayed satisfactory accuracy and precision. The assay recovery at low, medium and high concentrations was between 85 and 103% for both biomatrices, with a relative standard deviation of <15%. The lower limits of quantitation for plasma and mucosa were 100 ng/mL and 1.0 microg/mL, respectively. This method allowed measurement of PMF steady-state median concentrations in plasma (1.08 nmol/mL, n = 11; 10th and 90th percentiles: 0.633 and 2.385 nmol/mL) and mucosa (108.5 nmol/g, n = 9; 10th and 90th percentiles: 38.9 and 164.4 nmol/g) in mice which had received PMF (0.2%, w/w) with their diet.
Cai
Hong
H
Cancer Biomarkers and Prevention Research Group, Department of Cancer Studies and Molecular Medicine, University of Leicester, Leicester LE27LX, UK. hc47@le.ac.uk
Brown
Karen
K
Steward
William P
WP
Gescher
Andreas J
AJ
D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D005247:Feeding Behavior; D005419:Flavonoids / Q000032:analysis* / Q000097:blood / Q000737:chemistry; D006801:Humans; D007413:Intestinal Mucosa / Q000737:chemistry*; D051379:Mice; D008810:Mice, Inbred C57BL; D012680:Sensitivity and Specificity
C488726:5,7,3',4',5'-pentamethoxyflavone; D005419:Flavonoids; C017769:tricin