Development and Validation of Ultra Violet Spectrophotometric and Reversed-Phase High Performance Liquid Chromatography Techniques for Simultaneous Estimation of Brinzolamide and Brimonidine Tartrate in Ophthalmic Suspension Formulation

摘要

A sensitive and fast UV spectrophotometric technique and feasible reverse phase liquid chromatographic method for the analysis of Brinzolamide and Brimonidine tartrate in ophthalmic formulation has been developed. Linearity ranges for both methods were 5 - 25 Ig/ml and 1-5 ig/ml for brinzolamide and brimonidine tartrate respectively. The absorption maxima were observed at 232 and 257 nm for Brinzolamide and Brimonidine. The UV, ophthalmic formulation assay shows percentage purity ranging from 98.90 to 101.01% for Brinzolamide and 98.20 to 100.80% for Brimonidine. The mean percentage purity is 99.82% and 99.80% for Brinzolamide and Brimonidine respectively. The chromatographic retention time for Brinzolamide and Brimonidine was found to be 5.9 and 8.4 minutes respectively. The tailing factor was 0.808 and 0.860 for Brinzolamide and Brimonidine tartrate respectively. The method was validated according to the ICH guidelines and the method was found to be applicable for determination of Brinzolamide and Brimonidine tartrate from combined ophthalmic form.