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Pharmacokinetic studies of phloretin in beagle dogs plasma using LC–MS/MS

Libin W., Le M., Tian F., Xueying L., Shengyong Z.

Acta Chromatographica

2017

Vol. 29, no. 4

443--447

A simple, sensitive, and rapid liquid chromatography–tandem mass spectrometry (LC–MS/MS) method has been developed and validated for determination of phloretin in dog plasma using darunavir as internal standard. The phloretin was separated by the Inertsil® ODS3 C18 column (150 mm × 4.6 mm, 5 μm) and determined by LC–MS/MS. The electrospray ionization (ESI) source was operated in negative ionization mode for phloretin and positive ionization mode for darunavir (internal standard, IS). The multiple reaction monitoring (MRM) transitions were chosen to be m/z 273.0 → m/z 148.9 for phloretin, m/z 443.2 → m/z 401.0 for 2′,4′,6′,4-tetra-acetylphloretin and m/z 548.1 → m/z 69.1 for IS. The method was validated for accuracy, precision, linearity, range, selectivity, lower limit of quantification (LLOQ), recovery, and matrix effect. All validation parameters met the acceptance criteria according to regulatory guidelines. 2′,4′,6′,4-Tetra-acetylphloretin, as a prodrug of phloretin, is more stable than phloretin (PH) in vitro, protecting phenolic hydroxy from being oxygenated. The method had been successfully applied to a pharmacokinetic study of administration of phloretin and 2′,4′,6′,4-tetra-acetylphloretin in beagle dogs. Significant differences of tmax, Cmax, and area under the plasma concentration curve (AUC) were observed between phloretin and 2′,4′,6′,4-tetra-acetylphloretin.