Synthesis of the flavonoid 3',4',5'-trimethoxyflavonol and its determination in plasma and tissues of mice by HPLC with fluorescence detection.
3',4',5'-Trimethoxyflavonol (TMFol) was synthesized as a potential colorectal cancer chemopreventive agent. An HPLC method for determination for TMFol in murine plasma and tissues was developed and validated using human plasma. Analyte was separated (C(18) column; fluorescence detection 330nm excitation, 440nm emission) using 69% methanol and 0.1M ammonium acetate buffer (pH 5.1) as mobile phase. The method was linear for 50-2500ng/ml plasma and 0.05-10microg/g tissue (r>0.99). TMFol was recovered from plasma or tissues using solid phase columns or organic solvent protein precipitation, respectively. Recovery at low, medium and high concentrations was 97.6-107.3%, with inter- and intra-day coefficients of variation of <10%. The lower limit of quantitation for plasma was 50ng/ml. The method was applied to measure steady-state TMFol plasma and tissue levels in mice which received dietary TMFol (0.2%).
Britton
Robert G
RG
Department of Cancer Studies and Molecular Medicine, RKCSB, Leicester Royal Infirmary, University of Leicester, Leicester, LE2 7LX, UK.
Fong
Isabel
I
Saad
Shaban
S
Brown
Karen
K
Steward
William P
WP
Gescher
Andreas
A
Sale
Stewart
S
D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D005419:Flavonoids / Q000032:analysis* / Q000097:blood / Q000138:chemical synthesis*; D006801:Humans; D007668:Kidney / Q000737:chemistry*; D008099:Liver / Q000737:chemistry*; D051379:Mice; D008810:Mice, Inbred C57BL; D012680:Sensitivity and Specificity; D013050:Spectrometry, Fluorescence / Q000379:methods*
D005419:Flavonoids