Modeling retention and selectivity as a function of pH and column temperature in liquid chromatography

In reversed-phase liquid chromatography (RPLC), the retention of weak acids and bases is a sigmoidal function of the mobile-phase pH. Therefore, pH is a key chromatographic variable to optimize retention and selectivity. Furthermore, at an eluent pH close to the pKa of the solute, the dependence of ionization of the buffer and solute on temperature can be used to improve chromatographic separations involving ionizable solutes by an adequate handling of column temperature. In this paper, we derive a general equation for the prediction of the retentive behavior of ionizable compounds upon simultaneous changes in mobile-phase pH and column temperature. Four experiments, two limiting pH values and two temperatures, provide the input data that allow predictions in the whole range of these two variables, based on the thermodynamic fundamentals of the involved equilibria. Also, the study demonstrates the significant role that the choice of the buffer compound would have on selectivity factors in RPLC at temperatures higher than 25 °C. © 2006 American Chemical Society. Fil: Gagliardi, Leonardo Gabriel. Universidad de Barcelona; España. Provincia de Buenos Aires. Gobernación. Comisión de Investigaciones Científicas. Centro de Investigaciones en Tecnología de Pinturas. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Investigaciones en Tecnología de Pinturas; Argentina Fil: Castells, Cecilia Beatriz Marta. Universidad Nacional de La Plata; Argentina. Provincia de Buenos Aires. Gobernación. Comisión de Investigaciones Científicas. Centro de Investigaciones en Tecnología de Pinturas. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Investigaciones en Tecnología de Pinturas; Argentina