DEVELOPMENT AND VALIDATION OF ANALYTICAL METHOD FOR SIMULTANEOUS ESTIMATION OF BRINZOLAMIDE AND TIMOLOL BY HPLC FROM OPHTHALMIC PREPARATION

摘要

A fast, sensitive and accurate reverse phase liquid chromatographic method has been developed and validated for the simultaneous determination of Brinzolamide and Timolol maleate from ophthalmic preparation. Chromatographic separation of these two drugs were achieved on Zorbax Eclipse Plus, Agilent Technology (150mm x 4.6mm, 5μm) as stationary phase with a mobile phase comprising of triethylamine phosphate buffer : Acetonitrile : Methanol (70:20:10(v/v) at a flow rate of 1.0ml / min and PDA detection was set at 274 nm. Injection volume was 10 μl. Column oven temperature was maintained at 40°C. The retention time of timolol, and brinzolamide were 4.0 (±0.5) min, and 10.5 (±0.5) min respectively. Theoretical plate for Brinzolamide and Timolol were 5832 and 4876 respectively. The tailing factor was 1.09 and 1.21 for Brinzolamide and Timolol respectively. Resolution between the two compounds was 16.12. The developed method was validated according to the ICH guidelines and the proposed method can be applied for the routine quality control analysis of Brinzolamide and Timolol maleate from combined dosage form.