Stability-Indicating HPLC Method Development And Validation For Quantitative Analysis Of Oteseconazole - A Novel Antifungal Agent

Abstract

This study presents the development and validation of a stability-indicating Reverse Phase High Performance Liquid Chromatography (RP-HPLC) method for the quantification of Oteseconazole. The separation was performed using a Waters Alliance e2695 system with an Agilent Eclipse column (150 x 4.6 mm, 3.5 µm) and a mobile phase consisting of Ammonium Acetate (pH 3.0) and Acetonitrile (ACN) in a 30:70% v/v ratio, at a flow rate of 1.0 mL/min. Detection of the analyte was achieved using a photodiode array detector at 223 nm and ambient temperature. The drug exhibited a retention time of 3.365 minutes. Comprehensive method validation was conducted, assessing specificity, linearity, accuracy, precision, limit of detection (LOD), limit of quantification (LOQ), robustness, and degradation studies. Linear responses were observed for Oteseconazole within the range of 35.50 - 225.00 µg/mL (r² = 0.99984). The relative standard deviations for both interday and intraday precision were less than 2.0 %. The findings indicate that all method parameters comply with ICH guidelines for method validation. Additionally, the method was employed for stability studies through forced degradation experiments, revealing that the purity threshold remained below the purity angle across all conditions. The percentage degradation observed for acid, alkali, peroxide, reduction, thermal, photolytic, and hydrolysis conditions were 12.2 %, 11.6 %, 14.3 %, 4.0 %, 4.2 %, 2.1 %, and 1.7 %, respectively. Therefore, the developed RP-HPLC method is suitable for the quantification of Oteseconazole in routine pharmaceutical operations and analytical laboratories.