The influence of the memory effect on preparative separations using the amylose tris(3,5-dimethylphenylcarbamate) stationary phase

Acid/base mobile phase modifiers affect enantioseparations in ways that are not yet understood for the lack of systematic studies, which makes the scale-up of preparative separations difficult to predict. Shifts of the selectivity of certain pairs of enantiomers upon exposure of the column to these modifiers is amply documented. Furthermore, once the modifier has been removed from the mobile phase, the improved selectivity remains, this phenomenon has been named the memory effect. We selected four enantiomeric pairs for a systematic study of this memory effect. The selectivity of 4-chlorophenylalanine ethyl ester (4CPEE) improves after a solution of ethanesulfonic acid (ESA) is percolated through the column. The selectivity of propranolol HCl and Tröger's base increases after a solution of diiospropylethylamine is percolated through the column. The selectivity of these three pairs of enantiomers is inversely affected by percolation of the opposite acid/base solution. Each of these four compounds reached an equilibrium concentration that maintained the separation of the enantiomeric pairs. In contrast, the selectivity of trans-stilbene oxide (TSO) is not affected by either acid/base modifier. Preparative separations can be used to detect changes in the active surface of the chiral polymer stationary phase by measuring the change in selectivity and resolution when modifiers are used. Preparative method development was carried out on analytical columns and scale-up to 1 cm ID columns were performed in this study.