Method Development and Validation of Brinzolamide and Brimonidine In Its Bulk And Ophthalmic Dosage Form By Using RP-HPLC

Dr. K. Nageswara Rao^1*, Raghava Doonaboyina², R.Hema^3
1Professor and Head, Department of Pharmaceutical Analysis, K.G.R.L College of Pharmacy, KGRL Road, Bhimavaram, West Godavari, Andhra Pradesh, India.
²Associate Professor and Head, Department of Pharmaceutical Chemistry, K.G.R.L College of Pharmacy, KGRL Road, Bhimavaram, West Godavari, Andhra Pradesh, India.
³K.G.R.L College of Pharmacy, KGRL Road, Bhimavaram, West Godavari, Andhra Pradesh, India.

A B S T R A C T
In RP-HPLC method, the conditions were optimized to obtain an adequate separation of eluted compounds. Initially, various mobile phase compositions were tried, to separate title ingredients. Mobile phase and flow rate selection was based on peak parameters (height, tailing, theoretical plates, capacity or symmetry factor), run time and resolution. The mobile phase containing mixture of Phophate buffer solution: Methanol (65:35v/v, pH 4) with a flow rate of 1.0 ml/min is quite robust.The optimum wavelength for detection was 260 nm at which better detector response for both the drugs was obtained. The retention times for Brinzolamide and Brimonidine tartrate was found to be 2.113 min and 3.560 min, respectively. To ascertain its effectiveness, system suitability tests were carried out on freshly prepared stock solutions. The calibration was linear in concentration range of 5 to 25 μg/ ml and 20 to 100 μg/ml, with regression 0.9979 and 0.9999, Brinzolamide and Brimonidine tartrate respectively. The low values of % R.S.D indicate the method is precise and accurate. The mean recoveries were found above 99.3 % for both the drugs.
Keywords: Brinzolamide, Brimonidine tartrate, RP-HPLC, Validation.