Abstract

Abstract Background: A LCMS-MS method has been validated for the determination of naproxen using ketoprofen as an internal standard. Methods: Protein precipitate method was used to extract both analytes and internal standard from human plasma. Detection was made at m/z 229.1 / 169.0 for naproxen, m/z 253.08 / 209.0 for internal standard using esi positive ionization mode. Mass hunter 4.1 workstation software was used for the quantification. The stationary phase was Gemini C18, 50 x 4.6, 5μm. The optimum mobile phase was found to be acetonitrile and ammonium acetate buffer 20 mM (1:1 v/v). The separation was carried out by using temperature at 30°C (Degree centigrade) with a flow rate of 0.8 mL (milliliter). The injection volume was 5 L (microliter) and run time was 3 minutes. The retention time of analyte and internal standard was 2.054 and 1.922 minutes. Results: The method chosen is fast, robust, and sensitive. Each sample requires less than 3 min run time. The assay method is also highly specific due to the intrinsic selectivity of tandem mass spectrometry. The precision and accuracy are within the recommended limits in this concentration range. Conclusion: The developed method validated for accuracy, precision, linearity and recovery. The calibration curves were linear from 1.167 μg/mL to 163.729 μg/mL (microgram/ milliliter) for naproxen. The validated technique has been effectively applied to analyze human plasma samples for application in pharmacokinetic studies.

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