High-performance liquid chromatography: a comprehensive review

High-Performance Liquid Chromatography (HPLC) is a type of column chromatography that is generally used in biochemistry and analysis to separate, identify, and quantify active components. It is the most versatile, safest, reliable, and fastest chromatographic technique for the quality control of medicine factors. HPLC technique development and validation serve critical roles in novel drug discovery, development, and manufacturing, as well as a variety of other human and animal investigations. This article was prepared with an aim to review different aspects of HPLC, such as principle, types, instrumentation and application. Despite its widespread use, challenges related to method development, optimization of separation conditions, detector selection, and matrix interference remain significant. This review consolidates recent advancements and methodological strategies to address these challenges, thereby contributing to a clearer understanding of HPLC applications across pharmaceutical, environmental, food, and agricultural sciences.