Chemistry
Chemistry
10th Edition
ISBN: 9781305957404
Author: Steven S. Zumdahl, Susan A. Zumdahl, Donald J. DeCoste
Publisher: Cengage Learning
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In case of SEC method, the proteins’ adsorption could be a result of overall ionic interactions leading to lengthier retention time and peak asymmetry.True/False

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In the size exclusion chromatography (SEC), the factor the that differentiates one type of molecule from the other is the size of the molecules. As the sample is passed through the column that contain the packed material of stationary phase, due to the difference in size of the molecules, the smaller molecules get easily diffused  and adsorbed into the pores of the packing material in the column, while the larger particles are eluted out of the column with shorter retention time.

In certain cases, when the amino group of the protein has charge on them, that is positive or negative charge, and when the stationary phase of the column has packed material that are oppositely charged, it forms an attraction between the protein and stationary phase of the column. This affects the retention time of the protein, since it spends more time in the column due to the attraction between the oppositely charged ion. The interaction between the sample and the column material affects the peaks, its symmetry and its width. Therefore, ionic interaction causes uneven peak symmetry.      

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