共 25 条
Phosphorothioate oligonucleotides separation in ion-pairing reversed-phase liquid chromatography: effect of temperature
被引:22
作者:

Kadlecova, Zuzana
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Charles Univ Prague, Fac Sci, Dept Phys & Macromol Chem, Hlavova 8, Prague 12800, Czech Republic Charles Univ Prague, Fac Sci, Dept Phys & Macromol Chem, Hlavova 8, Prague 12800, Czech Republic

Kalikova, Kveta
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Charles Univ Prague, Fac Sci, Dept Phys & Macromol Chem, Hlavova 8, Prague 12800, Czech Republic Charles Univ Prague, Fac Sci, Dept Phys & Macromol Chem, Hlavova 8, Prague 12800, Czech Republic

Tesarova, Eva
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Charles Univ Prague, Fac Sci, Dept Phys & Macromol Chem, Hlavova 8, Prague 12800, Czech Republic Charles Univ Prague, Fac Sci, Dept Phys & Macromol Chem, Hlavova 8, Prague 12800, Czech Republic

Gilar, Martin
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Waters Corp, 34 Maple St, Milford, MA 01757 USA Charles Univ Prague, Fac Sci, Dept Phys & Macromol Chem, Hlavova 8, Prague 12800, Czech Republic
机构:
[1] Charles Univ Prague, Fac Sci, Dept Phys & Macromol Chem, Hlavova 8, Prague 12800, Czech Republic
[2] Waters Corp, 34 Maple St, Milford, MA 01757 USA
关键词:
Phosphorothioate oligonucleotides;
Ion-pairing reversed-phase chromatography;
Diastereomeric separation;
Temperature;
Resolution;
SENSITIVITY;
REAGENTS;
COLUMNS;
D O I:
10.1016/j.chroma.2022.463473
中图分类号:
Q5 [生物化学];
学科分类号:
071010 ;
081704 ;
摘要:
Analysis of diastereomers of phosphorothioate oligonucleotides in ion-pairing reversed-phase liquid chro- matography 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 degrees C to 90 degrees C) on retention, diastereomeric separation, resolution of mers of different length and reso- lution of oligonucleotides with different number of phosphorothioate linkages. It was observed that ele- vated 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.(c) 2022 Elsevier B.V. All rights reserved.
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