A simple HPLC method for the determination of apigenin in mouse tissues.
The flavone apigenin occurs in many leafy vegetables and fruits. It has been reported to have cancer chemopreventive efficacy in rodents. An HPLC method described previously for the determination of tricin, the dimethoxy cogener of apigenin, was modified and validated for measurement of apigenin in mouse tissues. Separation was carried out on a Hypersil-BDS C(18) column (4.6 x 250 mm) with an isocratic mobile phase of 55% methanol in 0.1 m ammonium acetate, pH 5.10, containing 0.27 mm disodium ethylenediamine tetraacetic acid. UV detection was at 336 nm, without interference from endogenous tissue compounds. The assay was linear in the range 25-400 ng/mL, 0.25-4 microg/mL and 2.5-40 microg/mL, with r(2) > 0.99 in all cases, for mouse plasma, liver and intestinal mucosa, respectively. Apigenin in mouse plasma, liver and intestinal mucosa was efficiently extracted with 0.1 m acetic acid in acetone. The assay recovery at low, medium and high concentrations was between 94.6 and 131.7% for all biomatrices, with a relative standard deviation of <10%. The lower limit of quantification for plasma was 25 ng/mL with a relative standard deviation of <15%. The method was used to measure the steady-phase apigenin levels in tissues of mice receiving apigenin in their diet.
Cai
Hong
H
Cancer Biomarkers and Prevention Research Group, Department of Cancer Studies and Molecular Medicine, University of Leicester, Leicester LE2 7LX, UK. hc47le.ac.uk
Raynaud
Delphine
D
Steward
William P
WP
Gescher
Andreas J
AJ
D000818:Animals; D047310:Apigenin / Q000032:analysis* / Q000097:blood / Q000737:chemistry; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D007413:Intestinal Mucosa / Q000378:metabolism*; D008099:Liver / Q000378:metabolism*; D051379:Mice; D015394:Molecular Structure; D015203:Reproducibility of Results
D047310:Apigenin