Impact of column selection for the analysis of 3mer phosphorothioated oligonucleotides in hydrophilic interaction liquid chromatography
The result's identifiers
Result code in IS VaVaI
<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00216208%3A11310%2F25%3A10504279" target="_blank" >RIV/00216208:11310/25:10504279 - isvavai.cz</a>
Result on the web
<a href="https://verso.is.cuni.cz/pub/verso.fpl?fname=obd_publikace_handle&handle=ksy1hMMXKM" target="_blank" >https://verso.is.cuni.cz/pub/verso.fpl?fname=obd_publikace_handle&handle=ksy1hMMXKM</a>
DOI - Digital Object Identifier
<a href="http://dx.doi.org/10.1016/j.chroma.2025.466404" target="_blank" >10.1016/j.chroma.2025.466404</a>
Alternative languages
Result language
angličtina
Original language name
Impact of column selection for the analysis of 3mer phosphorothioated oligonucleotides in hydrophilic interaction liquid chromatography
Original language description
With advancing development of therapeutic oligonucleotides, the demand for chromatographic methods for analysis and identification of oligonucleotide impurities and metabolites increases. 3mer oligonucleotides selected for this study represent common metabolites of nusinersen (drug Spinraza), in terms of length, sequence and phosphorothioate modifications. The aim of this work is to investigate the hydrophilic interaction liquid chromatography applicability to analysis of short 3mer oligonucleotides. Their ability to suppress diastereomeric separation and to resolve oligonucleotides differing in one nucleobase at different position in the sequence (3'end, 5'end, middle) was evaluated. The effects of the 3mer sequence and chromatographic conditions (pH of the aqueous part of the mobile phase, buffer concentration, column temperature) on the oligonucleotides' separation according to their sequence ("sequence selectivity") and peak broadening due to diastereomeric separation were investigated for selected zwitterionic columns, diol column, amide column, and two chiral columns (cyclofructan-and quinine-based). The zwitterionic stationary phases showed a great potential for analysis of short oligonucleotides. Diol column showed partial separation of diastereomers, interfering with the sequence-based oligonucleotides resolution due the peak broadening under all tested conditions.
Czech name
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Czech description
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Classification
Type
J<sub>imp</sub> - Article in a specialist periodical, which is included in the Web of Science database
CEP classification
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OECD FORD branch
10406 - Analytical chemistry
Result continuities
Project
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Continuities
I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace
Others
Publication year
2025
Confidentiality
S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů
Data specific for result type
Name of the periodical
Journal of Chromatography A
ISSN
0021-9673
e-ISSN
1873-3778
Volume of the periodical
1761
Issue of the periodical within the volume
October
Country of publishing house
NL - THE KINGDOM OF THE NETHERLANDS
Number of pages
7
Pages from-to
466404
UT code for WoS article
001586015700003
EID of the result in the Scopus database
2-s2.0-105016847850