Chromatography

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3 CHROMATOGRAPHY Factors That Impact Band Broadening Factor Impact Term impacted Especially important at Reasons Smaller Dm Smaller H B-term Decreased broadening due to longitudinal diffusion Larger H Cm -term Low mobile phase velocities High mobile phase velocities Smaller Ds Larger H Cs -term High mobile phase velocities Smaller d or df Smaller H Cs -term High mobile phase velocities Smaller dp Smaller H A-term High mobile phase velocities Smaller H Cm -term High mobile phase velocities Increased broadening due to less averaging of velocities because of fewer radial transfers between different mobile phase velocity paths.

Thus, for the entire collection of molecules within the zone, there is an overall average axial velocity at which the zone travels through the column. As with any process dictated by random movements, like diffusion, most of the molecules travel at or close to the average velocity, while a few travel faster and a few slower. The molecules that move slightly faster elute from the column at slightly shorter retention times, those that travel slightly slower elute at longer times, and most of them elute around the center of the peak.

C) Solutes in the mobile phase continue down the column and diffuse to reduce the radial concentration gradient. Similarly, solutes in the stationary phase diffuse out in response to the low concentration in the center of the column that arises because the mobile phase has pushed the rest of the molecules down the column. (d) The molecules that have reentered the mobile phase will now be pushed down the column, while molecules out in front are retained as they approach and interact with the stationary phase.

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