High Performance Liquid Chromatography: Basics

This lecture introduces the basics of High Performance Liquid Chromatography (HPLC), a method used for separating molecules based on their molar mass. HPLC is preferred over column chromatography for its improved selectivity and resolution, especially for thermally unstable or non-volatile molecules. The lecture covers the principles of HPLC chromatography, including the role of eluents, pumps, injectors, and chromatographic columns. It explains the factors influencing solute retention, such as column temperature, particle diameter, and eluent polarity. Different modes of elution, isocratic and gradient, are discussed, along with their impact on separation efficiency and resolution.