Chromatography: Comprehensive Study Document
1. Introduction to Chromatography
Chromatography is a laboratory technique used for the separation, identification, and
quantification of the components of a mixture. It involves passing a mixture dissolved in a
"mobile phase" through a stationary phase, where different components of the mixture move at
different rates, allowing for separation.
2. Principles of Chromatography
Chromatography operates based on two phases:
Stationary Phase: A fixed phase that interacts with the components of the mixture.
Mobile Phase: A fluid that carries the mixture through the stationary phase.
The separation occurs because each component of the mixture interacts differently with the
stationary and mobile phases, leading to different migration rates.
3. Types of Chromatography
Chromatography can be broadly categorized based on the states of the phases and the
mechanism of separation.
3.1 Liquid Chromatography (LC)
Principle: In LC, the mobile phase is a liquid, while the stationary phase is typically a solid or a
liquid on a solid support.
Applications: Used for separating polar and non-volatile compounds.
Types of Liquid Chromatography:
High-Performance Liquid Chromatography (HPLC): Involves high pressure to push the
mobile phase through a column packed with the stationary phase.
Size-Exclusion Chromatography (SEC): Separates molecules based on their size.
Ion-Exchange Chromatography (IEC): Separates ions and polar molecules based on charge.
