HPLC method development for simultaneous estimation of combined Amoxicillin Trihydrate/Flucloxacillin Sodium Capsules

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MAY, 2016
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A combined formulation of amoxicillin, a broad-spectrum penicillin and flucloxacillin, a narrow-spectrum beta-lactam antibiotic provides a wider spectrum of bactericidal activity against a wide range of gram-positive and gram-negative bacteria. An HPLC method was developed for the quantitative analysis of this combined formulation using methanol and acetate buffer of pH 5.0, in a ratio of 60:40; flow rate of 1.1ml/min and UV detection at 254nm, a Phenomenex C-18 column; 300 × 3.9; 10micron. The linearity of the method was assessed with a calibration graph with a correlation coefficient, r2 values of 0.9986 and 0.9967 for Flucloxacillin and Amoxicillin respectively. The LOD computed for Amoxicillin and Flucloxacillin were 0.0938 and 0.140 respectively. The LOQ obtained were 0.2843 and 0.3156 for Amoxicillin and Flucloxacillin respectively. The %RSD was estimated for three concentrations 0.64mg/ml, 0.8mg/ml and 0.96mg/ml by determining the %recovery for both Amoxicillin and Flucloxacillin. The %recovery for Amoxicillin for the concentrations stated were 100.67%, 99.88%, and 99.95%. That of Flucloxacillin were 99.78%, 99.96%, and 100.44%. The %RSD was also used to analyze the intra-day and inter-day precision. The intra-day for three different concentrations, 2mg/ml, 0.8mg/ml and 0.1mg/ml for Amoxicillin were 0.55, 1.38, and 0.93. That of Flucloxacillin were 0.19, 0.27, and 0.79. The %RSD for inter-day precision showed 0.92, 1.04, and 0.78 for Amoxicillin while that of Flucloxacillin were 0.40, 1.02, and 0.92. The method can be used to assay any Amoxicillin/Flucloxacillin combined dosage forms.
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Dissertation submitted to The Department of Pharmaceutical Chemistry of The Faculty of Pharmacy and Pharmaceutical Sciences, KNUST in partial fulfilment of the requirements for award of Master of Philosophy Degree in Pharmaceutical Chemistry
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