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  • Title: Determination of oseltamivir carboxylic acid in human serum by solid phase extraction and high performance liquid chromatography with UV detection.
    Author: Bahrami G, Mohammadi B, Kiani A.
    Journal: J Chromatogr B Analyt Technol Biomed Life Sci; 2008 Mar 15; 864(1-2):38-42. PubMed ID: 18313995.
    Abstract:
    This study was aimed at developing a fast and sensitive method for determination of oseltamivir carboxylic acid (OCA), the active moiety of anti-influenza agent, oseltamivir phosphate, in human serum by high performance liquid chromatography and UV detection. The analyte and an internal standard (vanillin) were extracted from human serum by a solid phase extraction (SPE) procedure. Chromatographic separation was achieved using a reverse phase C18 column with a mobile phase consisting of 0.05M phosphate buffer containing triethylamine (1mL/L; pH 3.0) and acetonitrile (70:30, v/v). The detection wavelength was set at 215nm. The average recoveries of the drug and internal standard were 98 and 85%, respectively. The calibration curve was linear over a concentration range of 15-6400ng/mL of OCA in human serum. The lower limits of detection and quantification were 5 and 15ng/mL, respectively. The coefficient variation values of both inter- and intra-day analysis were less than 12% whereas the percentage error was less than 4.5. The stability of the drug at the serum samples maintained at -40 degrees C for 60 days was found to be 100% from the initial value and no interferences were found from either endogenous components in serum or commonly co-administrated antiviral drugs. The validated method was applied to a randomized cross-over bioequivalence study of two different oseltamivir phosphate preparations in 24 healthy volunteers.
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