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

Md. Saniya, Sreelatha Gangu*, Mallani Sirisha, Sarvepalli Gnana Samanvitha, Shaik Naziya Fathima, Arunabha Mallik

ABSTRACT

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 an alytical 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 stu dies specific to antibiotics. It examines a wide range of chromatographic c onditions 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 stab ility indicating analytical methods to comply with ICH regulations and ensure the safety, efficacy, and shelf life of antibiotic formulations.

Keywords: RP HPLC, Antibiotics, Method Development, Validation, Stability Indicating Methods, Forced Degradation , ICH Q1A(R2), ICH Q2(R2), impurity profiling,