Reverse Phase High-performance Liquid Chromatography: A Comprehensive Review of Principles, Instrumentation, Analytical Procedures, and Pharmaceutical Applications
Suman Basak, Piyali Das
Abstract
Abstract A popular analytical method in the domains of material sciences, pharmacology, biology, the environment, food, cosmetics, and forensics is reverse phase high-performance liquid chromatography (RP-HPLC). Both polar and nonpolar molecules may be separated, identified, and quantified with its excellent resolution, reproducibility, and sensitivity. For improved retention and selectivity, polar drug analysis usually uses C18 columns, pH-controlled buffers, and aqueous-rich mobile phases. In contrast, nonpolar pharmaceuticals need a greater organic solvent content to guarantee solubility and efficient interaction with the hydrophobic stationary phase. Food additive analysis, the standardization of herbal products, biomarker measurement, drug stability research, therapeutic drug monitoring, pollutant detection, forensic toxicology, and biopharmaceutical characterization are among the uses of RP-HPLC. Even while RP-HPLC is quite versatile, it has drawbacks such as excessive solvent consumption, matrix effects, and trouble studying particularly big or very hydrophilic macromolecules. Optimizing chromatographic settings and carefully preparing samples are necessary to overcome these constraints. With ongoing advancements in technology, RP-HPLC continues to be a fundamental component of contemporary analytical science, providing reliable results for both simple tests and intricate research requirements.