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Unformatted text preview: by pouring
acid into the column
acid a. A continuous flow of solvent carries a solution of solutes A and B
down a column. (a)
b. As the solvent carries the two solutes down the column, we begin to
see some separation of the solution occurs. (b)
c. At some later point in time, it can be seen that solute B is moving at a
much faster rate than A. (c)
In (d), solute B emerges first, while solute A finally emerges in (e).
Thus, solute A has a greater affinity for the stationary phase than solute
B. By varying the pH of the solvent or temperature of the column, the
output of the column can be significantly altered, such as the timing of
when individual species emerge. The Chromatogram
Different peaks on the chromatogram
correspond to different components in the
The chromatograms shows the results of
separations of protein mixtures by ion exchange
chromatography. The lettered peaks correspond
to different proteins (A = ovalbumin, B =
conalbumin, C = cytochrome c, D = lysozyme).
The separation corresponding to the
chromatogram on the left was performed at pH
5.85, while the one on the right was performed
at pH 6.5.
It is evident that operation conditions such as pH
and temperature have a significant effect on the
The level of complexity of the sample is
indicated by the number of peaks which appear.
Qualitative information about the sample
composition is obtained by comparing...
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- Spring '14