A brief Introduction to HPLC and its principle

Introduction of HPLC, instrumentation and principle

1. Chromatography

2. Chromatography is a laboratory technique used for separating mixtures into their individual components. The basic principle of chromatography involves passing a mixture dissolved in a "mobile phase" through a "stationary phase." Different substances in the mixture interact differently with the stationary phase, leading to their separation. The separation is based on the differential affinities of components for the stationary and mobile phases. Components that have a higher affinity for the stationary phase will move more slowly, while those with higher affinity for the mobile phase will move faster.

3. Types of Chromatography Gas Chromatography (GC) Liquid Chromatography (LC) (includes HPLC) Adsorption Chromatography Partition Chromatography Ion-Exchange Chromatography Size-Exclusion Chromatography Affinity Chromatography Paper Chromatography Thin-Layer Chromatography (TLC) Column Chromatography

4. HPLC HPLC stands for High-Performance Liquid Chromatography , a powerful analytical technique used to separate, identify, and quantify components in a mixture. It operates by passing a liquid sample through a column packed with a solid adsorbent material under high pressure. The different components of the sample interact with the adsorbent material differently, causing them to travel through the column at different speeds, leading to their separation.

5. Components of HPLC Pump : Delivers the solvent (mobile phase) through the column at high pressure. Injector : Introduces the liquid sample into the mobile phase. Column : Filled with a stationary phase (usually a solid or gel), where the separation takes place. Detector : Monitors the separated components as they exit the column and generates a signal for data analysis. Data system : Collects and analyzes the data generated by the detector

6. How HPLC Works Animation Link: https://www.youtube.com/watch?v=Vr5t-cgHHG4