Rp-HPLC in Modern Pharmaceutical Analysis: From Fundamental Principles to Method Validation and Emerging Trends

Narender Boggula

Narender Boggula Associate Professor Department of Pharmaceutical Chemistry Omega College of Pharmacy, Edulabad, Ghatkesar, Hyderabad - 501301, Telangana, India.

Abstract

Reversed-phase high-performance liquid chromatography (RP-HPLC) is one of the most widely used analytical techniques for the separation, identification, and quantification of pharmaceutical compounds. In RP-HPLC, a non-polar stationary phase, commonly C18-bonded silica, is used with a relatively polar mobile phase, generally consisting of water or buffer and an organic modifier such as methanol or acetonitrile. Separation primarily depends on the hydrophobic interactions between analytes and the stationary phase, while factors such as pH, ionic strength, organic-solvent composition, flow rate, and column temperature influence retention and selectivity. RP-HPLC offers high resolution, sensitivity, reproducibility, and versatility and is therefore extensively employed in pharmaceutical quality control, assay determination, impurity profiling, dissolution testing, stability studies, and biological sample analysis. Method validation is an essential component of pharmaceutical RP-HPLC and includes characteristics such as specificity, accuracy, precision, linearity, range, detection limit, and quantitation limit according to the intended analytical purpose. Despite its advantages, RP-HPLC has limitations including high solvent consumption, relatively expensive instrumentation, column deterioration, and the generation of chemical waste. Recent developments involving analytical quality by design (AQbD), shorter columns, greener solvents, and improved detection systems are contributing to more efficient and sustainable RP-HPLC methods. This review briefly discusses the principles, advantages, disadvantages, applications, and future significance of RP-HPLC in pharmaceutical analysis.

Keywords: RP-HPLC, reversed-phase chromatography, pharmaceutical analysis, C18 column, method validation, quality control

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