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 degrees C.
Gagliardi
Leonardo G
LG
Departament de Química Analítica, Facultat de Química, Universitat de Barcelona, Barcelona, Spain.
Castells
Cecilia B
CB
Ràfols
Clara
C
Rosés
Martí
M
Bosch
Elisabeth
E