Introduction: Timolol maleate and Brinzolamide are used to treat eye diseases such as angular hypertension and glaucoma (open angle type). These two drugs work together to lower fluid levels within the eye which in turn helps to reduce the pressure inside the eye and consequently prevents eye diseases such as blindness and glaucoma. Objectives: This work has developed simple, reproducible, accurate, and precise analytical method employing the RP-HPLC technique for the simultaneous identification and quantification purpose of ophthalmic suspension of Brinzolamide and Timolol maleate and Brinzolamide for ocular use. Methodology: The two medications were separated in one run with the use of a UV detector set at 274 nm using an AD HOC Prudent C18 (125 mm x 4.6 mm; 5µ) P/N:P18-12540-5 column with isocratic & acetonitrile and 70:30% (v/v) 0.1 M acetate buffer solution pH 4.25 as a mobile phase. The flow rate was fixed at 1.0 mL/min. Results: Timolol and Brinzolamide are two drugs that have retention times of 1.7 and 2.5 minutes respectively and their resolution is more than 2. The recovery percentage for both Timolol and Brinzolamide was within the limit range of 98% to 102%. As indicated by the relevant USP chapter <1225> and by ICH guideline (Q2-R2) that was suggested, the approach that has been designed and described was effective, reliable and accurate. Conclusion: The given method will be useful in QC laboratory for process and finished product study of ophthalmic solution useful in glaucoma treatment.