Unique green chromatographic method for the qualitative and quantitative analysis of ketoconazole, its impurities and preservatives from ketoconazole cream formulation: Identification of degradants by Q-ToF LCMS and Robustness by Design of Experiments

Abstract

Current research portrays a simultaneous determination method for the qualitative and quantitative analysis of Ketoconazole, its impurities and preservatives from Ketoconazole cream formulation. The unique method was developed using the RP-UPLC technique with a gradient elution mode. The separation of isomeric preservative components was achieved on the Waters Acquity UPLC BEH C18 column using phosphate buffer as mobile phase A and a mixture of phosphate buffer and acetonitrile, 30:70 (v/v) as mobile phase B. The set flow rate, injection volume, column oven temperature, and detector wavelengths were 0.5 mL/min, 2.0 μL, 30 °C, and 225 nm, respectively. The method's stability indicating nature was achieved through forced degradation studies. The active drug was prone to acidic and oxidative conditions. The attained acidic and peroxide degradant peaks were identified using Q-ToF LCMS. The developed method was validated per regulatory guidelines, and the data showed superior precision with a %RSD of 0.7 for Ketoconazole, 0.8 for preservatives, and 1.6 for impurities. All the components showed excellent linearity (r > 0.999) within the 50–150% range for Ketoconazole and preservatives and from LOQ to 150% for impurities. The mean accuracy data resulted from 99.4% to 99.9% for Ketoconazole, 99.9%–106.0% for impurities, and 100.3%–101.4% for butylated hydroxyanisole. The robust method conditions were evaluated by the Design of Experiments. The greenness and eco-friendliness of the method were demonstrated by modern tools such as the analytical eco scale, GAPI and AGREE. The final UPLC method is specific and suitable for determining the preservatives, drug content and impurities of Ketoconazole simultaneously from its cream formulation, which saves analysis time and solvent consumption supporting towards the green chemistry principles.