Development and Validation of RP-HPLC Method for the Simultaneous Estimation of Tafluprost and Timolol in Combined Tablet Dosage Form

Abstract

A simple, rapid, accurate, and precise Reverse Phase High-Performance Liquid Chromatography (RP-HPLC) method was developed and validated for the simultaneous estimation of Tafluprost and Timolol in combined tablet dosage form. Tafluprost is a prostaglandin F2α analogue used to reduce intraocular pressure, while Timolol is a non-selective beta-adrenergic receptor blocker widely used in the treatment of glaucoma. Accurate and reliable analytical methods are essential to ensure the quality, safety, and efficacy of such combination pharmaceutical products.

Chromatographic separation was achieved using a C18 column with a suitable mobile phase composition under optimized conditions. The analytes were detected at an appropriate wavelength where both drugs exhibited adequate absorbance. The developed method provided well-resolved peaks with satisfactory retention times, good peak symmetry, and acceptable system suitability parameters.

The method was validated according to ICH guidelines for specificity, linearity, accuracy, precision, limit of detection (LOD), limit of quantification (LOQ), and robustness. The calibration curves were linear within the selected concentration range with correlation coefficients close to 1. The percentage recovery values indicated high accuracy, and low %RSD values confirmed good precision. The method proved to be robust against small deliberate variations in chromatographic conditions.

The validated RP-HPLC method was successfully applied for routine quantitative analysis of Tafluprost and Timolol in combined tablet dosage form and can be effectively used for quality control and stability studies.