Tunable normal phase enantioselectivity of amino acid esters via mobile phase composition

Abstract

The ability to tune chiral selectivity through mobile phase modifiers is a powerful tool in chiral separations. Beyond improving efficiency and/or resolution, some mobile phase systems can even invert elution order, a highly desirable result for trace analyses or preparative scale isolations. Previous work has demonstrated that acidic modifiers, such as ethanesulfonic acid (ESA), can greatly impact separations of enantiomers. However, prior studies were primarily performed on coated chiral stationary phases (CSPs), which limited the selection of the bulk mobile phase component.In this work, the effect of ESA modifier was studied for the enantioseparation of six pairs of amino acid esters on a CHIRALPAK® IA column, an immobilized amylose-based CSP, with different combinations of standard solvents (hexane and ethanol) as well as “non-standard” solvents, such as methyl t-butyl ether, ethyl acetate, tetrahydrofuran, acetone, or 1,4-dioxane. ESA generally improved selectivity, and multiple instances of elution order reversal were observed. A Van Deemter plot study reveals that ESA exerts its effect by pulling the enantiomer deeper into the chiral cavity of the chiral polymer to increase the interactions between the analytes and the stationary phase, which is the main reason for the increased enantioselectivity.