Combined effects of mobile phase composition and temperature on the retention of phenolic antioxidants on an octylsilica polydentate column

• Phenolic antioxidants were separated at 60–100 °C on a polydentate C8 column.
• The temperature effects on retention are well described by the linear van’t Hoff model.
• A simple retention model describes combined effects of temperature and mobile phase.
• Molecular size and solute hydrogen-bond acceptor capacity control the retention.
• High-temperature solvent gradients are superior to temperature programming.

Combined effects of temperature and mobile-phase composition on retention and separation selectivity of phenolic acids and flavonoid compounds were studied in liquid chromatography on a polydentate Blaze C8 silica based column. The temperature effects on the retention can be described by van’t Hoff equation. Good linearity of ln k versus 1/T graphs indicates that the retention is controlled by a single mechanism in the mobile phase and temperature range studied. Enthalpic and entropic contributions to the retention were calculated from the regression lines. Generally, enthalpic contributions control the retention at lower temperatures and in mobile phases with lower concentrations of methanol in water. Semi-empirical retention models describe the simultaneous effects of temperature and the volume fraction of the organic solvent in the mobile phase. Using the linear free energy-retention model, selective dipolarity/polarizability, hydrogen-bond donor, hydrogen-bond acceptor and molecular size contributions to retention were estimated at various mobile phase compositions and temperatures. In addition to mobile phase gradients, temperature programming can be used to reduce separation times.