Characterization of three macrocyclic glycopeptide stationary phases in supercritical fluid chromatography

Abstract

Macrocyclic glycopeptides have been used as chromatographic stationary phases for over twenty years, particularly for their ability to separate enantiomers. While they are mostly used with buffered aqueous liquid mobile phases, they can also be used in supercritical fluid chromatography (SFC) with mobile phases comprising pressurized carbon dioxide and a co-solvent (like methanol), possibly comprising acidic or basic additives.In the present study, we compared three macrocyclic glycopeptide stationary phases (Chirobiotic V2, Chirobiotic T and Chirobiotic TAG) in SFC with carbon dioxide – methanol (90:10) containing no additives. First, the interactions contributing to retention are evaluated with a modified version of the solvation parameter model, comprising five Abraham descriptors (E, S, A, B, V) and two additional descriptors to take account of interactions with ionizable species (D− and D+). Linear solvation energy relationships (LSER) are established based on the retention of 145 achiral analytes.Secondly, the contributions of interactions to enantioseparations are discussed, based on the analysis of 67 racemates. The individual success rate on each phase was observed to be moderate, especially as these phases are known to be more efficient when acidic or basic additives are employed. Chirobiotic TAG proved more successful than the other two phases. Discriminant analyses were computed to gain some insight on retention mechanisms, but only Chirobiotic TAG provided interpretable results.Finally, the effects of a small proportion of acidic or basic additive on enantioseparation with Chirobiotic T stationary phase are briefly discussed.