Riboflavin Binding Protein Chiral Stationary Phase: Investigation on the Retention Mechanism.

The chiral recognition mechanism of Riboflavin Binding Protein (RfBP) stationary phase has been investigated by studying the influence of temperature on the retention process. The effect of temperature on retention and resolution of two analytes was studied between 15°C and 55°C and van't Hoff plots of lnk and ln versus 1/T were acquired. The thermodynamic parameters were calculated and the data were used to derive some information regarding the retention mechanism in this chromatographic system. Quantitative structure-enantioselective retention relationships (QSERR) were derived by multiple regression analysis using various molecular descriptors and the retention data determined on RfBP-stationary phase. The results were interpreted in terms of structural requirements of the specific binding site on the biomacromolecule to provide an insight of the recognition mechanism.