Journal of Chromatography Open (Nov 2024)
Comparison of the adsorption dynamics of leucyl-leucine enantiomers and glycyl-glycine on Chirobiotic R and Chirobiotic V columns
Abstract
The adsorption dynamics of the LL- and DD-enantiomers of Leu-Leu and Gly-Gly on a Chirobiotic V column packed with a CSP bearing glycopeptide antibiotic vancomycin was studied and compared with the adsorption dynamics of the same dipeptides on a Chirobiotic R column packed with a CSP with bonded antibiotic ristocetin A. The column efficiencies were essentially different. The heights equivalent to a theoretical plate (HETP) on Chirobiotic R were 2-4 times larger than on Chirobiotic V, and the shape of van Deemter plots, strongly convex-upwards on the former column, demonstrated minor (Leu-Leu) or lacking (Gly-Gly) convexity on the latter column. This difference is attributed to differences in the binding kinetics of dipeptides to the antibiotics, fast for vancomycin and slow for ristocetin A. Since the kinetic C term of the van Deemter equation cannot explain such a large discrepancy in HETP for the same analyte on two columns, and since the A and B terms for the Chirobiotic R column were essentially larger than for the Chirobiotic V column, it is suggested that binding kinetics influences eddy and longitudinal diffusion processes associated with those terms. It is also hypothesized that unusually high values of van Deemter B coefficients occasionally found in chiral chromatography are not experimental artifacts but the manifestation of coupling the effects of slow adsorption kinetics and longitudinal diffusion.