Entropic-based separation of diastereomers. Size-exclusion chromatography with on-line viscometry and refractometry detection for analysis of blends of mannose and galactose methyl-alpha-pyranosides at "ideal" size-exclusion conditions

Published

January 11, 2021

Author(s)

Andre M. Striegel, Steven Trainoff

Abstract

TThe separation of carbohydrate diastereomers by an ideal size-exclusion mechanism, i.e., in the absence of enthalpic contributions to the separation, can be considered one of the grand challenges in chromatography: Can a difference in the location of a single axial hydroxy group on a pyranose ring (e.g., the axial OH being located on carbon 2 versus on carbon 4 of the ring) sufficiently affect the solution conformational entropy of a monosaccharide in a manner which allows for members of a diastereomeric pair to be separated from each other by size-exclusion chromatography (SEC)? Previous attempts at answering this question, for aqueous solutions, have been thwarted by the mutarotation of sugars in water. Here, the matter is addressed by employing the non-mutarotating methyl-alpha-pyranosides of D-mannose and D-galactose. We show for the first time, using SEC columns, the entropically-driven separation of members of this diastereomeric pair, at a resolution of 1.2 to 1.3 and with only a 0.4 % to 1 % change in solute distribution coefficient over a 25 oC range, thereby demonstrating the ideality of the separation. It is also shown how the newest generation of on-line viscometer allows for improved sensitivity, thereby extending the range of this so-called "molar-mass-sensitive" detector into the monomeric regime. Detector multidimensionality is showcased via the synergism of on-line viscometry and refractometry, which combine to measure the intrinsic viscosity and viscometric radius of the sugars continually across the elution profiles of each diastereomer, methyl-alpha-D-mannopyranoside and methyl-alpha-D- galactopyranoside.

Citation

Chromatographia

Volume

84

Pub Type

Journals

Download Paper

https://doi.org/10.1007/s10337-020-03983-z

Local Download

Keywords

Size-exclusion chromatography, viscometry, diastereomers, carbohydrates, refractometry

Citation

Striegel, A. and Trainoff, S. (2021), Entropic-based separation of diastereomers. Size-exclusion chromatography with on-line viscometry and refractometry detection for analysis of blends of mannose and galactose methyl-alpha-pyranosides at "ideal" size-exclusion conditions, Chromatographia, [online], https://doi.org/10.1007/s10337-020-03983-z (Accessed October 11, 2025)