The separation of components in a mixture based on their different distributive behaviors between a liquid mobile phase and a solid or liquid stationary phase.
The system consists of: Solvent reservoir, Pump, Injector, Column (stationary phase), Detector, and Data acquisition system (Integrator).
Because high pressure (typically 50 to 400 bar) is required to force the mobile phase through a column packed with very small particles (3–5 µm) to achieve high resolution.
The time interval between the point of injection and the appearance of the peak maximum at the detector.
A graphical representation of the detector response (y-axis) versus time (x-axis).
The Column, because that is where the actual separation takes place.
The total volume of mobile phase in the column; the volume required to elute a non-retained compound.
USP General Chapter <621>.
A porous filter at the column inlet/outlet that prevents stationary phase particles from escaping and blocks large contaminants from entering the column.
Isocratic: Uses a constant mobile phase composition throughout the run.
Gradient: Changes the composition over time to elute compounds with widely different polarities.
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