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Bioanalytical Method Development and Validation for Simultaneous Estimation of Cefixime and Dicloxacillin by RP-HPLC in Human Plasma

Somnath D. Bhinge, Sharangouda M. Malipatil, Lalit V Sonawane


An accurate, rapid and simple reversed-phase high performance liquid chromatography (RP-HPLC) bioanalytical method was developed and validated for simultaneous estimation of cefixime, dicloxacillin in human plasma using ezetimibe as an internal standard. The cefixime, dicloxacillin and internal standard were extracted by liquid–liquid extraction technique. Chromatographic separation is accomplished using CAPCELL PAK C18 (4.6 mm x 250 mm, 5 mm) analytical column. The mobile phase consisted of phosphate buffer, acetonitrile and methanol in 42:55:03 proportions. Detection and quantification were performed by UV/Vis detection at 225 nm. The lower limit of quantification was 0.5 μg mL–1 for both cefixime and dicloxacillin in human plasma. The calibration curves were linear over the concentration range 0.5 to 40 μg mL−1 for both drugs in human plasma. The method was quantitatively evaluated in terms of linearity, precision, accuracy, recovery, selectivity, and stability. The method was found to be simple, convenient and suitable for the analysis of cefixime and dicloxacillin from biological fluids.


Bioanalytical method; validation; human plasma; cefixime; dicloxacillin

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Copyright (c) 2014 Somnath D. Bhinge, Sharangouda M. Malipatil, Lalit V Sonawane

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