A simple and robust high-performance liquid chromatography coupled to a diode-array detector method for the analysis of genistein in mouse tissues.
A simple liquid-liquid extraction procedure and quantification by high-performance liquid chromatography (HPLC) method coupled to a diode-array detector (DAD) of genistein (GEN) was developed in various mouse biological matrices. 7-ethoxycoumarin was used as internal standard (IS) and peaks were optimally separated using a Kinetex C18 column (2.6μm, 150mm×2.10mm I.D.) at 40°C with an isocratic elution of mobile phase with sodium dihydrogen phosphate 0.01M in water at pH 2.5 and methanol (55:45, v/v), at a flow rate of 0.25mL/min. The injection volume was 10μL. In all cases, the range of GEN recovery was higher than 61%. The low limit of quantification (LLOQ) was 25ng/mL. The linearity of the calibration curves was satisfactory in all cases as shown by correlation coefficients >0.996. The within-day and between-day precisions were <15% and the accuracy ranged in all cases between 90.14% and 106.05%. This method was successfully applied to quantify GEN in liver, spleen, kidney and plasma after intravenous administration of a single dose (30mg/kg) in female BALB/C mice.
Department of Pharmacy and Pharmaceutical Technology, School of Pharmacy, University of Navarra, C/Irunlarrea 1, E-31008 Pamplona, Spain.
C
C
Tamames-Tabar
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E
Imbuluzqueta
M A
MA
Campanero
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P
Horcajada
M J
MJ
Blanco-Prieto
D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003374:Coumarins / Q000737:chemistry; D005260:Female; D019833:Genistein / Q000032:analysis* / Q000737:chemistry / Q000302:isolation & purification / Q000493:pharmacokinetics; D016018:Least-Squares Analysis; D059625:Liquid-Liquid Extraction; D051379:Mice; D008807:Mice, Inbred BALB C; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D014018:Tissue Distribution
D003374:Coumarins; C017299:7-ethoxycoumarin; D019833:Genistein
Biological matrices; Extraction method; Genistein; HPLC; Method validation