Question

How can molecules with same charge at varying amounts be separated by chromatography?

Isn't the answer is increasing the concentartion of the buffer slution? Please provide a good answer.

Answer 1

Chromatographic separation is the physical separation of components in a compound based on their affinity towards the stationary phase and mobile phase. All chromatographies follow the same principle, i.e., the compound having more affinity towards the mobile phase moves along with the mobile phase (gas or liquid) and the those that have more affinity towards the stationary phase (liquid or solid), move slower. So, to characterize a specific component in a compound or to isolate a component, chromatographic separation is necessary.

Molecules with the same charge at varying amounts can be separated by changing the pH or ionic strength of the eluting buffer, which enables the separation of molecules by minimizing the electrostatic forces.