Advances in High-Performance Liquid Chromatography (HPLC) and Ultra-Performance Liquid Chromatography (UPLC)

Abstract

High-Performance Liquid Chromatography (HPLC) and Ultra-Performance Liquid Chromatography (UPLC) are fundamental analytical techniques that have revolutionized chemical analysis across various fields. These chromatographic methods offer exceptional separation capabilities, high resolution, and precise quantification of complex mixtures. HPLC has evolved significantly since its inception, with improvements in column technology, detection systems, and automation. UPLC, introduced as an advancement to conventional HPLC, operates at significantly higher pressures using sub-2-μm particle sizes, resulting in enhanced resolution, speed, and sensitivity. Both techniques have found extensive applications in pharmaceutical analysis, environmental monitoring, food safety, clinical diagnostics, and biomedical research. The implementation of various detection methods, including UV-visible, fluorescence, mass spectrometry, and electrochemical detection, has expanded their analytical capabilities. Recent developments in column technology, such as core-shell particles and monolithic columns, have further improved separation efficiency. The integration of artificial intelligence and machine learning has enhanced method development and data analysis. This review discusses the fundamental principles, technological advancements, and diverse applications of HPLC and UPLC. Additionally, it discusses current challenges, emerging trends, and future prospects in chromatographic science, including green chromatography initiatives and miniaturization efforts. The continuous evolution of these techniques contributes significantly to analytical chemistry's advancement, promising even more sophisticated applications in various scientific disciplines.