Development of an HPLC Method Using Chemometric Approaches for the Determination of Latanoprostene Bunod in Pharmaceutical Preparations

Latanoprostene bunod has recently emerged as a promising ocular antihypertensive agent due to its potent ability to lower intraocular pressure and its favorable safety profile observed in clinical trials. In this study, a new, simple, and selective HPLC method was developed for quantifying latanoprostene bunod in pharmaceutical formulations. A Box-Behnken experimental design was employed to optimize the chromatographic parameters, including flow rate, organic solvent, and acid content in the mobile phase. The analysis was performed on an C18 column (4.6 × 50 mm, 3.5 µm), with a mobile phase consisting of water and acetonitrile, both containing 0.01% formic acid (45:55, v/v). Chromatographic conditions were set at a flow rate of 0.6 mL/min, a column temperature of 40 °C, and detection at 260 nm. Method validation, following the ICH Q2(R1) guideline, demonstrated excellent linearity over the concentration range of 0.50–20.00 µg/mL, with LOD and LOQ values of 0.052 µg/mL and 0.157 µg/mL, respectively. Precision and accuracy were confirmed with recovery rates between 100-102% and RSD values below 3%, respectively. The validated method was then successfully applied to the analysis of latanoprostene bunod in a commercially available ophthalmic formulation, confirming its suitability for routine quality control use.