Influence of temperature in reverse-phase high-performance liquid chromatography with gradient elution

The influence of temperature on the retention of several species separated by reverse-phase liquid chromatography by gradient elution is shown to be of enough importance to warrant careful control of temperature if reproducible results are to be obtained. The smaller the particle size in the column, the greater the effect of temperature, and therefore the control should be greater. Likewise, it has been verified that for a given solvent gradient, independent of its complexity, there is a linear relation between ln k′ and 1/T, which also occurs in separations by isocratic elution. Dufek's equation can be adjusted perfectly to the experimental data obtained from gradient elutions, and may be used in the simulation and optimization of gradient chromatographic processes.