Phosphorothioate oligonucleotides separation in ion-pairing reversed-phase liquid chromatography: effect of temperature.

Autor: Kadlecová Z; Department of Physical and Macromolecular Chemistry, Faculty of Science, Charles University, Hlavova 8, 12800, Prague, Czech Republic., Kalíková K; Department of Physical and Macromolecular Chemistry, Faculty of Science, Charles University, Hlavova 8, 12800, Prague, Czech Republic. Electronic address: kalikova@natur.cuni.cz., Tesařová E; Department of Physical and Macromolecular Chemistry, Faculty of Science, Charles University, Hlavova 8, 12800, Prague, Czech Republic., Gilar M; Waters Corporation, 34 Maple Street, Milford, MA 01757, United States of America. Electronic address: Martin_Gilar@waters.com.
Jazyk: angličtina
Zdroj: Journal of chromatography. A [J Chromatogr A] 2022 Oct 11; Vol. 1681, pp. 463473. Date of Electronic Publication: 2022 Sep 07.
DOI: 10.1016/j.chroma.2022.463473
Abstrakt: Analysis of diastereomers of phosphorothioate oligonucleotides in ion-pairing reversed-phase liquid chromatography is affected not only by the character and concentration of ion-pairing system, but also by the separation temperature. In this work, eight ion-pairing systems at two concentrations buffered with acetic acid were used with octadecyl column to investigate the effects of temperature (in the range from 20 °C to 90 °C) on retention, diastereomeric separation, resolution of mers of different length and resolution of oligonucleotides with different number of phosphorothioate linkages. It was observed that elevated temperature suppresses the diastereomeric separation and oligonucleotide peaks become narrower. This improves the resolution of n and n-1 mers at elevated temperature. Plots of ln k (k = retention factor) versus reciprocal absolute temperature show that for 100 mM ion-pairing systems the increase in temperature does not lead to simple decrease in oligonucleotides retention as generally observed in reversed-phase liquid chromatography. The aim of this work is to improve chromatographic method for analysis of phosphorothioate oligonucleotides.
Competing Interests: Declaration of Competing Interest The authors declared no conflict of interest.
(Copyright © 2022 Elsevier B.V. All rights reserved.)
Databáze: MEDLINE
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