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Abstract
Sensitive, simple and accurate thin-layer chromatography (TLC) - densitometry method for simultaneous determination of brinzolamide (BRZ) and timolol maleate (TML) in their pharmaceutical formulations was developed and validated, where separation was occurred using a mobile phase composed of chloroform: methanol: ammonia (9:0.5:0.1 by volume) on silica gel 60 F254 plates, densitometric detection was done at 254 nm for brinzolamide and 298 nm for timolol. The calibration curves are linear over the concentration range of 3-16 μg/spot for BRZ and 4-14 μg/spot for TML. The obtained results were statistically compared with those obtained by the reported methods, showing no significant difference. The developed method was validated according to the ICH guidelines and was successfully applied for simultaneous determination of brinzolamide and timolol maleate in bulk drug and in their ophthalmic preparation.