An accurate, precise, sensitive and relatively simple RP-HPLC method was developed for the simultaneous estimation of and brimonidine tartrate (BRT) and timolol maleate (TM) in pure form as well as in pharmaceutical formulation. The chromatographic separation was achieved on Spheri-5 RP-18 (25 cm×4.6 mm, 5 µ), perkin elmer: brownlee columns with a mobile phase combination of Acetonitrile: Buffer (0.1% triethylamine pH adjusted to 2.5 using o-phosphoric acid) (40:60 %v/v) at a flow rate of 1ml/min. UV Detection was carried out at 309 nm. Retention time for BRT was 3.2 min and that of TM was 4.1 min. The method was found to be linear in the range of 2-12 µg/ml for BRT and 5-30 µg/ml for TM. The LOD values for BRT and TM were found to be 0.59 µg/ml and 1.63 µg/ml respectively. The LOQ values were found to be 1.96 µg/ml and 4.5 µg/ml respectively for BRT and TM. The method was validated for accuracy, precision (intra-day and inter-day precision studies) and robustness as per the ICH guidelines and the %RSD values were found within the acceptable criteria (<2%).
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