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J Chromatogr A. 2013 Aug 23;1304:127-32. doi: 10.1016/j.chroma.2013.06.078. Epub 2013 Jul 4.

Reversed phase ion-pairing chromatography of an oligolysine mixture in different mobile phases: effort of searching critical chromatography conditions.

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  • 1Center for Biocatalysis and Bioprocessing of Macromolecules, The Polytechnic Institute of New York University, Six Metrotech Center, Brooklyn, NY 11201, USA.

Abstract

Our earlier study [J. Chromatogr. A 1218 (2011) 7765] on separation of an oligolysine mixture consisting of chains with 2-8 lysine residues (number of lysine residues, dp=2-8) by ion-pairing reversed-phase chromatography using heptafluorobutyric acid (HFBA) as an ion pairing reagent at fixed mobile phase acetonitrile (ACN) content was extended to isocratic elution conditions with different ACN percentages. The present work explored how manipulating the mobile phase HFBA concentration ([HFBA]) and %-ACN content influences separations of the oligolysine mixture. The closed pairing model was used to analyze variation of the retention factor as a function of [HFBA]. The partition coefficient of the paired peptide decreased with increasing %-ACN. Pairing of HFBA to oligolysine was cooperative, and the effect increased when %-ACN in the mobile phase was lowered. A plot of the partition coefficient as a function of %-ACN for oligolysines varying in dp converged at one ACN content, indicating a critical condition in which components of different dp co-elute.

Copyright © 2013 Elsevier B.V. All rights reserved.

KEYWORDS:

Critical chromatography condition; Heptafluorobutyric acid; Ion-pairing chromatography; Mobile phase composition; Oligolysine

PMID:
23876768
[PubMed - indexed for MEDLINE]
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