A NOVEL RP-HPLC-UV METHOD FOR SIMULTANEOUS DETERMINATION OF AMOXICILLIN/CLAVULANIC ACID IN HUMAN PLASMA
ANAB FATIMA *
Faculty of Pharmacy, Hamdard University, Karachi, Pakistan
SYED BAQIR NAQVI
Faculty of Pharmacy, Hamdard University, Karachi, Pakistan
JIBRAN– Bin- YOUSUF
Department of Pharmacy, Agha Khan University Hospital, Karachi, Pakistan
SHEIKH ABDUL KHALIQ
Faculty of Pharmacy, Hamdard University, Karachi, Pakistan.
SHAHEEN PERVEEN
Department of Pharmaceutics, Faculty of Pharmacy, Jinnah University for Women, Karachi, Pakistan
*Author to whom correspondence should be addressed.
Abstract
A simple, novel, responsive and cost-effective reversed phase high performance liquid chromatography (RP-HPLC-UV) method has been developed and validated for concurrent estimation of Co-amoxiclav (amoxicillin+clavulanic acid) in human plasma. The method was précised, reproducible and with greater accuracy. The assay method can apply for quantitative measurement of amoxicillin and clavulanic acid in therapeutic drug monitoring up to LOQ of 0.0037 µgml⁻¹ and 0.06 µgml⁻¹ respectively. The procedure was simple, one step extraction with complete analytical recovery (i.e. mean recovery % for amoxicillin for three conc. was found to be (MEAN ±SD) (%) 96.6±1.65, 99.8±0.22, 98.9±1.55 and for clavulanic acid it was 97.9±2.1, 99.5±0.61 and 97.9±1.06 respectively. In an isocratic mode separation was achieved using RP 18e column (250 x 4.6 cm). Mobile phase consist of methanol (10 volume) and 0.02 M phosphate buffer (90 volume) at flow rate of 1.3 ml/min. The eluent detection from column carried out at 235 nm. The percentage deviation for within day and between day assay was found to be less than 5% for both components.
Keywords: RP-HPLC-UV, plasma, amoxicillin, clavulanic acid, pharmacokinetic, validation