Development of a sensitive LC–MS/MS method for quantification of coniferyl ferulate and its metabolite coniferyl alcohol in rat plasma: Application to a pharmacokinetic study

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HIGHLIGHTSConiferyl ferulate and its metabolite coniferyl alcohol were simultaneously determined.Method validation procedures were in accordance with EMA regulations.It was successfully applied to the pharmacokinetic study in rats.A rapid and simple LC–MS/MS method was developed and validated for the simultaneous determination of coniferyl ferulate (CF) and its metabolite coniferyl alcohol (CA) using bavachromene as an internal standard (IS). A TSQ Quantum Access mass spectrometer was operated under selected-reaction monitoring mode using negative electrospray ionization. Extraction with ether was used in sample preparation. The plasma samples were prepared and then chromatographed on a Phenomenex Luna C18 column (2.1 mm × 50 mm, 1.7 μm; Torrance, USA) at 35 °C, using acetonitrile: water (65:35, v/v) in an isocratic mode at a flow rate of 0.3 mL/min. Method validation was performed as per the FDA guidelines and calibration curves showed good linearity over the concentration range of 2.5–1000 ng/mL for both CF and CA. The intra- and inter-day precision and accuracy were within the acceptable limits. The developed assay was successfully applied to a pharmacokinetic study of CA in rats.

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