Reversed-Phase High-Performance Liquid Chromatography (RP-HPLC) Based Analytical Method Development and Validation for Pharmaceutical Dosage Forms: A Comprehensive Review

Liquid chromatography forms the backbone of quality evaluation, regulatory compliance, and safety profiling in the pharmaceutical field. Out of the various available techniques, Reversed-Phase High-Performance Liquid Chromatography (RP-HPLC) is still the most widely used tool to identify and quantify active pharmaceutical ingredients (APIs) and their final dosage forms. This review takes a close look at the basic principles of RP-HPLC and provides a clear, step-by-step guide to developing reliable analytical methods. We practically evaluate key variables that affect separation, such as picking the right stationary phase, choosing suitable mobile phases and buffers, adjusting pH, and selecting the best detection systems. Beyond basic method development, this paper also dives into stability-indicating assays (SIAs) and forced degradation studies. These stress tests are crucial for understanding how drugs break down when exposed to heat, light, acid, base, and oxidation over time. Furthermore, we outline the current method validation requirements dictated by the updated International Council for Harmonization (ICH) Q2(R2) guidelines, breaking down parameters like specificity, linearity, precision, accuracy, detection limits, and robustness. To bridge the gap between traditional practices and modern trends, this review also touches upon the growing shift toward Analytical Quality by Design (AQbD) and Green Analytical Chemistry (GAC). Ultimately, this article aims to serve as a practical and comprehensive guide for laboratory analysts and researchers working in drug formulation and quality control.