Simultaneous estimation of cefixime and cloxacillin in human plasma by reversed phase-HPLC with UV detection

Abstract

An accurate, rapid and simple reversed-phase high performance liquid chromatography (RP-HPLC) method was developed and validated for simultaneous estimation of cefixime, cloxacillin and for chloramphenicol (internal standard) in human plasma. Chromatographic separation was accomplished using CAPCELL PAK C8 (4.6 mm × 250 mm, 5 μm) analytical column using phosphate buffer, methanol and acetonitrile (40:45:15) as eluent. Detection and quantification were performed by UV/Vis detection at 225 nm. The lower limit of detection and quantification were 0.01 μg mL-1 and 0.25 μg mL-1 for cefixime, 0.03 μg mL-1 and 0.39 μg†mL-1 for cloxacillin. The calibration curves were linear over the concentration range 0.5 to 40 μg mL-1 for both cefixime and cloxacillin in human plasma. The method was quantitatively evaluated in terms of linearity, precision, accuracy, recovery, selectivity, and stability. The method is simple, convenient and suitable for the analysis of cefixime and cloxacillin in human plasma.