Abstract
Background and the purpose of the study: To develop a simple, rapid and accurate HPLC method for simultaneous quantitative determination of Latanoprost, Timolol and Benzalkonium chloride (BAK) in ophthalmic solution. Method: Chromatographic separation was achieved with PDA detector using Inertsil C18, 300 x 3.9mm, 5µ reverse phase analytical column. The mobile phase consist of buffer: acetonitrile (40: 60 v/v), was passed through the column at flow rate of 1.0 ml/min. The method was performed at wavelength gradient .The experiment was carried out at 30oC. Results: The calibration curves were linear in the concentration range of 25% to 150% of the working concentration (r2 > 0.999). The lower limit of quantification was 0.8, 0.9 and 0.6 for Timolol, BAK and Latanoprost respectively. Conclusion: The developed procedure was used for simultaneous quantitative estimation of Latanoprost, Timolol and Benzalkonium chloride (BAK) in ophthalmic solution. Developed method was validated as per ICH Q2 (R1), and most useful for academic as well as industrial scale. Key words: Latanoprost, Timolol, Benzalkonium Chloride, RP HPLC, Validation.
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