Development, validation, and greenness assessment of UV spectrophotometric and RP-HPLC methods for the quantitative determination of Nafithromycin

Nafithromycin is a novel ketolide antibiotic approved for the treatment of community-acquired bacterial pneumonia (CABP). The present study describes the development and validation of UV spectrophotometric and reverse-phase high-performance liquid chromatographic (RP-HPLC) methods for the quantitative determination of Nafithromycin. UV analysis was performed at 275 nm, while RP-HPLC separation was achieved on a Phenomenex Kinetex ® C18 column using Methanol: 10 mM ammonium acetate buffer (pH 9.0) (70:30, v/v) as the mobile phase. The developed methods were validated according to ICH Q2 (R2) guidelines for specificity, linearity, precision, trueness, detection capability, robustness, assay, and solution stability. Both methods showed linear responses over the concentration range of 2–12 µg/mL with correlation coefficients greater than 0.999. Recovery values were within acceptable limits and %RSD values were below 2.0%, confirming method reliability. Assay of marketed Nafithromycin tablets yielded results within pharmacopeial acceptance criteria. Statistical comparison of assay results using one-way ANOVA demonstrated no significant difference between the two methods (F = 2.58, F critical = 4.96), indicating comparable analytical performance. The environmental sustainability of the methods was evaluated using Green Analytical Procedure Index (GAPI) and Analytical GREEnness Metric Approach (AGREE), which confirmed their compliance with green analytical chemistry principles. The proposed UV spectrophotometric and RP-HPLC methods are suitable for routine quality control analysis of Nafithromycin pharmaceutical formulations.