An accurate, precise and relatively simple High Performance Thin Layer Chromatographic Method was developed for the simultaneous estimation of Brimonidine Tartarte (BRT) and Timolol Maleate (TM) in pure form as well as in pharmaceutical formulation. Chromatographic Separation was carried out on TLC aluminium sheet pre-coated with silica gel G 60 F254 using Hexane: Iso-propyl alcohol : ammonia (5:2:0.5 v/v/v) as mobile phase. Detection was carried out densitometrically at 281 nm. The Rf value of BRT and TM was found to be 0.36 and 0.65 respectively. The method was validated for linearity, accuracy, precision, LOD and LOQ. The linearity range of Brimonidine Tartrate was to be between 200-600 ng/spot and that of Timolol Maleate was found to be 500-1500 ng/spot with regression coefficient values of 0.996 and 0.995 for Brimonidine Tartrate and Timolol Maleate respectively. The accuracy of the method was assessed by performing recovery studies at 80%, 100% and 120%. The % recovery for Brimonidine Tartrate was found to be 100.37-101.3% and that of Timolol Maleate was found to be 99.93-100.44%. The LOD and LOQ values for Brimonidine Tartrate were found to be 21.16 and 64.11 ng/spot respectively and Timolol Maleate were found to be 101.2 and 306.63 ng/spot respectively. Precision of the method was assessed by performing repeatability, intra-day and inter-day precision studies. Thus, the developed method was found to be accurate, precise and relatively simple for the simultaneous estimation of Brimonidine Tartrate and Timolol Maleate in pure form as well as in marketed formulation.
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