METHOD DEVELOPMENT AND VALIDATION OF ANTIBIOTICS BY RP-HPLC WITH EMPHASIS ON STABILITY STUDIES: A COMPREHENSIVE REVIEW

Antibiotics remain a cornerstone of global healthcare, yet increasing antimicrobial resistance and strict quality-control requirements necessitate highly accurate and robust analytical methods for their quantification. Reversed-phase high-performance liquid chromatography (RP-HPLC) is the most widely used analytical technique for the routine and stability-indicating analysis of antibiotic drug substances and formulations. This review provides a comprehensive overview of RP-HPLC method development strategies, validation requirements, and forced degradation studies specific to antibiotics. It examines a wide range of chromatographic conditions applied to β-lactams, macrolides, fluoroquinolones, Cephalosporins, tetracyclines, and aminoglycosides. A systematic literature review summarizes major analytical methods reported in the last two decades, highlighting chromatographic challenges, degradation pathways, impurity profiling, and method robustness considerations. Emerging analytical trends such as green chromatography, Quality by Design (QbD), and hyphenated techniques (PDA/MS) are also discussed. The review emphasizes the need for stability-indicating analytical methods to comply with ICH regulations and ensure the safety, efficacy, and shelf life of antibiotic formulations.