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Retention modeling under organic modifier gradient conditions in ion-pair reversed-phase chromatography. Application to the separation of a set of underivatized amino acids

Analytical and Bioanalytical Chemistry, ISSN: 1618-2642, Vol: 397, Issue: 6, Page: 2151-2159
2010
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The combined effect of the ion-pairing reagent concentration, C , and organic modifier content, φ, on the retention under φ-gradient conditions at different constant C was treated in this study by using two approaches. In the first approach, the prediction of the retention time of a sample solute is based on a direct fitting procedure of a proper retention model to 3-D φ-gradient retention data obtained under the same φ-linear variation but with different slope and time duration of the initial isocratic part and in the presence of various constant C values in the eluent. The second approach is based on a retention model describing the combined effect of C and φ on the retention of solutes in isocratic mode and consequently analyzes isocratic data obtained in mobile phases containing different C values. The effectiveness of the above approaches was tested in the retention prediction of a mixture of 16 underivatized amino acids using mobile phases containing acetonitrile as organic modifier and sodium dodecyl sulfate as ion-pairing reagent. From these approaches, only the first one gives satisfactory predictions and can be successfully used in optimization of ion-pair chromatographic separations under gradient conditions. The failure of the second approach to predict the retention of solutes in the gradient elution mode in the presence of different C values was attributed to slow changes in the distribution equilibrium of ion-pairing reagents caused by φ-variation. © 2010 Springer-Verlag.

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