Abstract

High-performance liquid chromatography (HPLC) is a powerful and widely used analytical technique for the separation, identification, and quantification of compounds in complex mixtures. This abstract provides an overview of HPLC, its principles, instrumentation, and applications. HPLC is based on the principle of chromatography, where a sample mixture is separated into its individual components using a stationary phase and a mobile phase. The stationary phase is typically a packed column with specific properties, while the mobile phase is a liquid solvent or a mixture of solvents that carries the sample through the column. The components in the sample interact differently with the stationary phase, leading to their separation. Instrumentation for HPLC typically includes a pump to generate high-pressure flow of the mobile phase, an injector for sample introduction, a column for separation, a detector to monitor the eluting compounds, and a data analysis system. Various detectors such as UV-Vis, fluorescence, and mass spectrometry can be employed for detection, enabling selective and sensitive analysis. HPLC finds wide-ranging applications in pharmaceutical analysis, environmental analysis, food and beverage analysis, biochemical analysis, and clinical diagnostics. It plays a crucial role in drug discovery, quality control, environmental monitoring, food safety assessment, and biomolecule analysis. HPLC techniques have evolved over the years, leading to advancements such as high-resolution HPLC and hyphenated techniques like liquid chromatography-mass spectrometry (LC-MS). In conclusion, HPLC is a versatile and indispensable analytical technique that offers high separation efficiency, selectivity, and sensitivity. Its ability to separate and quantify diverse compounds makes it an invaluable tool in various scientific disciplines and industries. Ongoing advancements continue to enhance its capabilities and broaden its applications, driving further advancements in the field of analytical chemistry.