HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY (HPLC) METHOD DEVELOPMENT AND VALIDATION: A COMPREHENSIVE REVIEW

Pranjal Dattu Avhad* and Madhuri Bhausaheb Nagare

ABSTRACT

High-Performance Liquid Chromatography (HPLC) is a highly efficient analytical technique extensively employed in pharmaceutical, environmental, and food industries for the separation, identification, and quantification of chemical compounds. The accuracy and reliability of HPLC methods are crucial for ensuring consistent quality control in drug formulation and other applications. This review explores the fundamental principles of HPLC, the stepwise approach to method development, and the critical aspects of method validation in compliance with International Council for Harmonisation (ICH) guidelines. The method development process involves optimizing various chromatographic parameters, including mobile phase composition, column selection, flow rate, and detection techniques. Additionally, method validation ensures robustness and reproducibility through parameters such as accuracy, precision, specificity, linearity, limit of detection (LOD), limit of quantification (LOQ), robustness, and ruggedness. The significance of HPLC in pharmaceutical analysis, including drug stability testing, impurity profiling, and bioanalytical studies, is also discussed. Advancements in chromatographic techniques continue to improve analytical performance, making HPLC a critical tool for quality assurance and regulatory compliance in pharmaceutical sciences.

Keywords: HPLC, Method Development, Validation, ICH Guidelines, Analytical Techniques.