The bandwidth in gradient elution chromatography with a retained organic modifier

The variance of a chromatographic band is derived in the case of RPLC gradient elution when the organic modifier is significantly retained onto the stationary phase. This derivation is based on the extension of a model due to Poppe et al. [H. Poppe, J. Paanakker, M. Bronckhorst, J. Chromatogr., 204 (1981) 77] which assumes that the gradient front remains unchanged and propagates along the column at the same speed as the mobile phase, following piston flow. Theoretical and experimental results are compared in the case of caffeine on a C1-silica stationary phase eluted with an acetonitrile gradient. The actual retention behaviors of caffeine and acetonitrile were implemented in the theoretical calculations. The model predicts compression factors between 0.71 and 0.34 for relatively smooth gradient steepness, βt0βt0, between 0.009 and 0.054 while the corresponding experimental band compression factors vary between 1.01 and 0.43 for the very same gradient steepness. The model underestimation of these factors arises likely from the strong deviation of the actual retention behavior from the prediction of the Linear Solvent Strength Model (LSSM).