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A RP-HPLC method was developed for estimation of 5-Hydroxy methyl furfural content in Levofloxacin Hemihydrate intravenous infusion 5mg/ml. It involved a 150mm x 4.6mm, 5µm LichroSphere C-18 column with 40°C temperature. The separation was achieved on simple isocratic method. A mixture of pH 3.0 buffer (0.04M, ortho phosphoric acid buffer, adjusted pH 3.0 with triethylamine) - acetonitrile (87:13, v/v) was used as the mobile phase. The flow rate was 1.0ml/min, injection volume was 5µl and the detection wavelength was 284nm. The retention time of 5HMF was 2.7min. The total runtime was 12min within which active compound and degradation products were separated. The developed method successfully applied to the determination of 5HMF content in pharmaceutical preparations. The developed RP-HPLC method was validated with respect to linearity, accuracy, specificity, limit of detection, limit of quantification, precision, robustness and ruggedness.
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