METHOD DEVELOPMENT AND VALIDATION OF SIMULTANEOUS ESTIMATION OF AZITHROMYCIN AND DEXAMETHASONE IN EYE DROPS BY RP-HPLC

A simple, reversed phase high performance liquid chromatography (RP-HPLC) method has been developed for the simultaneous estimation of Azithromycin and Dexamethasone in their combined formulation from the eye drop. The GRACE ODS C18, 250 x 4.6 mm, 5 μm, L1 packing, column was maintained at an ambient temperature and 230 nm λ max conditions. The mixture of Methanol and 0.0335M Phosphate Buffer (pH 7.5) in the ratio of (80:20 v/v) mobile phase was used in the flow rate of 1.2 ml/min. Azithromycin and Dexamethasone peaks were eluted at 3.832 min and 4.798 min respectively with good resolution. The above method was validated in terms of System suitability, linearity, accuracy, precision, Limit of Detection (LOD), Limit of Quantification (LOQ) in accordance with ICH guidelines. The results obtained from the method validation were within the acceptable limit. In conclusion a method has been developed for the simultaneous estimation of Azithromycin and Dexamethasone in combined dosage form. The linearity range of Azithromycin and Dexamethasone were found to be from 0.1-12μg/ml for Azithromycin, 0.1-8μg/ml for Dexamethasone. Linear regression coefficients were found to be 0.9931 and 0.9949 for Azithromycin and Dexamethasone respectively.