A Critical Review of RP-HPLC Methods for Antibiotic Agents: Emphasis on Metronidazole and Tinidazole

Reverse Phase High-Performance Liquid Chromatography (RP-HPLC) has become a gold-standard technique in pharmaceutical analysis owing to its accuracy, reproducibility, and versatility. Among antibiotic agents, Metronidazole and Tinidazole are important nitroimidazole derivatives widely prescribed for bacterial and protozoal infections. This review carefully compiles and evaluates reported RP-HPLC methods for estimating Metronidazole and Tinidazole in bulk drugs, pharmaceutical dosage forms, and biological samples. The analysis covers method development strategies, chromatographic conditions (mobile phases, stationary phases, flow rates, and detection wavelengths), and validation parameters in line with ICH guidelines. Literature shows various RP-HPLC approaches for the single and simultaneous determination of Metronidazole and Tinidazole. Differences in mobile phase, detector choice, and column specifications directly influence sensitivity, linearity, and resolution. Stability-indicating methods and bioanalytical applications further enhance the usefulness of RP-HPLC in addressing pharmaceutical and clinical needs. The review highlights advantages, limitations, and emerging trends, such as eco-friendly solvent systems and hybrid analytical techniques. RP-HPLC remains the preferred method for analyzing Metronidazole and Tinidazole due to its robustness and regulatory acceptance. Compiling existing methods provides valuable guidance for researchers and industry professionals seeking to optimize analysis procedures. Future developments are likely to focus on green chromatography, high-throughput analysis, and the combination of techniques for improved sensitivity and efficiency.