Development of a sensitive LC-MS/MS method for quantification of coniferyl ferulate and its metabolite coniferyl...

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 (2.1mm×50mm, 1.7μm; Torrance, USA) at 35°C, using acetonitrile: water (65:35, v/v) in an isocratic mode at a flow rate of 0.3mL/min. Method validation was performed as per the FDA guidelines and calibration curves showed good linearity over the concentration range of 2.5-1000ng/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.