The objective was to develop and validate suitable HPTLC method for simultaneous determination of Timolol maleate and Bimatoprost in ophthalmic dosage form. Materials and Methods: Chromatographic separation was carried out on TLC plate pre-coated with silica gel 60F254 using Ethyl acetate: Toluene: Methanol: Formic acid (3.5: 3.0: 1.5: 0.1 v/v/v/v) as mobile phase and scanning the plate at 260 nm. The Rf value for Timolol maleate and Bimatoprost were 0.30 and 0.54 respectively. The method was linear in the concentration of 3417.70-17088.50 ng/band for Timolol maleate and 150-750 ng/band for Bimatoprost with a correlation coefficient of 0.998 and 0.999 for Timolol maleate and Bimatoprost respectively. The percentage recoveries obtained for Timolol and Bimatoprost in the range of 99.31-101.39% and 99.80-101.23% trespectively. The result of analysis have been validated as per the International conference on Harmonisation (ICH) guidelines. The developed methods adequately separated both the drugs. Validation results indicated that the method shows satisfactory linearity, accuracy, precision and found to be specific. So, the proposed methods can be used successfully for routine analysis of Timolol maleate and Bimatoprost in ophthalmic dosage form by HPTLC.
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