Development and Validation of Stability Indicating RP-LC Method for Estimation of Related Substances of Enrofloxacin in Bulk and Its Pharmaceutical Formulations

An isocratic reverse phase liquid chromatography (RP-LC) method has been developed and subsequently validated for the determination of Enrofloxacine and its related substances in Bulk and its pharmaceutical tablet formulation. Separation was achieved with a Inertsil ODS 3V (150 mmx4.6 mm I. D; particle size 3 mu m)) Column and 1% orthophosphoric acid buffer (pH adjusted to 2.5 with triethyl amine): acetonitrile (800: 200) v/v as eluent at a flow rate of 1.0 mL/min. UV detection was performed at 270nm. The method is simple, rapid, and selective. The described method was validated as per ICH guidelines for specificity, forced degradation, linearity and ruggedness. Known and unknown impurities are well resolved from each other and also degradation products. all the known impurities shown a linear responses with very good LOD and LOQ values below 0.05%. The % Recoveries for all the impurities lies within the ICH limits. The results showed that the proposed method is suitable for the precise, accurate and rapid determination of related substances of enrofloxacin in bulk, its dosage forms.