Quantification of fenoprofen in human plasma using UHPLC–tandem mass spectrometry for pharmacokinetic study in healthy subjects |
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Authors: | Kirtikumar D Bharwad Priyanka A Shah Pranav S Shrivastav Vinay S Sharma Puran Singhal |
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Institution: | 1. Department of Chemistry, School of Sciences, Gujarat University, Ahmedabad, Gujarat, India;2. Department of Chemistry, Faculty of Basic and Applied Science, Madhav University, Sirohi, Rajasthan, India;3. Bioanalytical Department, Alkem Laboratories Ltd., Lower Parel, Mumbai, Maharashtra, India |
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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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Keywords: | fenoprofen fenoprofen-d3 human plasma pharmacokinetic study solid-phase extraction UHPLC–MS/MS |
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