Ozenoxacin (OZ) is a novel synthetic quinolone antimicrobial agent that has been recently approved by US-FDA for the treatment of impetigo in combination with its preservative, benzoic acid (BA). Meanwhile, implementing greener chromatography has been a recent goal of a great ecological importance. In the proposed research, two novel green methods have been validated for the simultaneous determination of OZ and BA in skin creams for the first time. An organic-solvent free mixed-micellar HPLC technique was developed using RP-C8 core-shell column with a mobile phase consisting of 0.01 M Brij-35, 0.15 M sodium dodecyl sulfate, and 0.02 M ammonium acetate in water, adjusted to pH 4.5. The mobile phase flow rate was 1 mL min−1 and detecting analytes in short retention times (3.4 and 4.7 min for BA and OZ, respectively) at wavelength 235 nm. The working concentration ranges were 1–10 and 10–100 μg mL−1 for BA and OZ, respectively. Another eco-friendly high-performance thin-layer chromatography (HPTLC) method was designed using HPTLC silica-gel 60 F254 plates as a stationary phase and ethyl acetate–ethanol–ammonia (6:4:2, v/v/v) as mobile phase. The detection was also performed at λmax = 235 nm. The method was found linear across concentration range of 0.5–1.8 and 0.05–0.18 μg/band for OZ and BA, respectively which gave symmetric, compact band with Rf values of 0.21 and 0.69 for OZ and BA, respectively. The proposed methods were applied successfully in their combined marketed pharmaceutical cream. The proposed methods were typically based on safe biodegradable reagents, and their ecological superiority was proven using recent greenness metrics.
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