Clinical Chemistry 5 Chromatography and Electrophoresis

A continuous flow of solvent carries a solution of

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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 sample mixture. 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 output. Information 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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