Quantification of fenoprofen in human plasma using UHPLC–tandem mass spectrometry for pharmacokinetic study in healthy subjects

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Abstract
A rapid, simple and sensitive ultra‐performance liquid chromatography–tandem mass spectrometry (UHPLC–MS/MS) method has been developed to quantify fenoprofen, a nonsteroidal anti‐inflammatory drug in human plasma for a pharmacokinetic study in healthy subjects. Owing to high levels of protein binding, protein precipitation followed by solid‐phase extraction was employed for the extraction of fenoprofen and fenoprofen‐d3 (used as internal standard) from 200 μL human plasma. Separation was performed on a BEH C18 (50 × 2.1 mm, 1.7 μm) column using methanol−0.2% acetic acid in water (75:25, v/v) under isocratic elution. Electrospray ionization was operated in the negative mode for sample ionization. Ion transitions used for quantification in the selected reaction monitoring mode were m/z 241/197 and m/z 244/200 for fenoprofen and fenoprofen‐d3, respectively. Under the optimized conditions, fenoprofen showed excellent linearity in the concentration range 0.02–20 μg/mL (r2 ≥ 0.9996), adequate sensitivity, favorable accuracy (96.4–103.7%) and precision (percentage coefficient of variation ≤4.3) with negligible matrix effect. The validated method was successfully applied to a pharmacokinetic study of fenoprofen in healthy subjects. The significant features of the method include higher sensitivity, small plasma volume for processing and a short analysis time.

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