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How does chromatography separate biological compounds?
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Biochemistry
In biochemistry, researchers study the chemical substances and processes that occur in living organisms and cells. In particular, various processes by which cells gain energy, such as epinephrine-stimulated cAMP synthesis, have been deduced through the study of biochemistry.
Chromatography separates biological compounds by distributing them between two phases: a moving fluid, known as the mobile phase, and a contiguous stationary phase.1
The separation relies on the principle of differential migration. As a mixture is transported through the system by the mobile phase, each component interacts differently with the stationary phase based on its specific chemical or physical properties. Components with a higher affinity for the stationary phase are retained longer, while those with a lower affinity migrate more rapidly with the mobile phase.12
Common mechanisms used to achieve this separation include:
- Adsorption: Solute molecules bond directly to the surface of the stationary phase.3
- Partition: Solutes distribute based on their relative solubility in a liquid stationary phase.3
- Size-Exclusion: Molecules are separated based on their size as they diffuse into and out of the porous structure of the stationary phase.34
- Affinity: Exploits specific biological interactions, such as enzyme-substrate or antigen-antibody binding, to selectively retain target molecules.34
- Ion-Exchange: Separates molecules based on their ionic charge.4