Analytical Method Development and Validation for the Determination of Brinzolamide and Brimonidine Using Reverse Phase HPLC Method in Bulk and Pharmaceutical Dosage Form

Dr R. Vani¹*, Amena Samreen², Humera Tabassum³, Dr M.sunitha⁴*
¹Head of Department, Pharmaceutical Analysis and QA, Shadan Women’s College of Pharmacy, Hyderabad.
²Post-Graduate Student, Shadan Women’s College of Pharmacy, Hyderabad.
³Post-Graduate Student, Shadan Women’s College of Pharmacy, Hyderabad.
⁴Principal, Shadan Women’s College of Pharmacy, Hyderabad.

ABSTRACT
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 2.137 min and 2.844 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 to100 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.