High-performance Liquid Chromatography

High-performance liquid chromatography (HPLC) is a powerful analytical technique used for separating, identifying, and quantifying components in complex mixtures. This chapter provides an in-depth exploration of HPLC's fundamental principles, instrumentation, key characteristics, and applications, especially in forensic analysis. HPLC separates compounds based on interactions between the stationary phase and mobile phase. Two key principles that govern this separation are partitioning and adsorption. The instrumentation used in HPLC consists of several components: a column filled with the stationary phase, a precision pump to deliver the mobile phase at high pressure, an injector to introduce samples into the mobile phase, and a detector (e.g., UV-Vis, fluorescence) to monitor the eluting compounds. HPLC is characterized by high pressure, which enables efficient separations, and theoretical plates, which measure the column's efficiency. Its key advantages include high resolution with efficient separations, sensitivity with low detection limits, and versatility with various column and detector options. The HPLC is widely used for the analysis of non-volatile organic compounds, including drugs, pesticides, food, and plant products.