Kaczmarkiewicz Anna, Zielak Judyta, Nuckowski Łukasz, Studzińska Sylwia
Chair of Environmental Chemistry and Bioanalytics, Faculty of Chemistry, Nicolaus Copernicus University 7 Gagarin Str. PL-87-100 Toruń Poland
RSC Adv. 2019 Nov 28;9(67):39100-39110. doi: 10.1039/c9ra06483d. eCollection 2019 Nov 27.
The main goal of this study was the investigation of the impact of several ionic liquids, commonly used as free silanol suppressors, on the retention and separation of phosphorothioate oligonucleotides. Three various stationary phases (octadecyl, octadecyl with embedded polar groups and pentafluorophenyl) as well as ionic liquids with the concentration range of 0.1-7 mM were used for this purpose. The results obtained during this study showed that the increase in concentration of ionic liquids results in increasing retention of the oligonucleotides. Such an effect was observed regardless of the stationary phase used. Moreover, elongation of the alkyl chain in the structure of ionic liquids caused an increase of antisense oligonucleotide retention factors. The results obtained during retention studies confirmed that addition of ionic liquids to the mobile phase influences antisense oligonucleotide retention in a way similar to the case of commonly used ion pair reagents such as amines. A method of oligonucleotide separation was also developed. The best selectivity was obtained for the octadecyl stationary phase since separation of mixtures of antisense oligonucleotides and their metabolites differing in sequence length was successful. It has to be pointed out that ionic liquids were used for the first time as mobile phase additives for oligonucleotide analysis.
本研究的主要目标是考察几种常用作游离硅醇抑制剂的离子液体对硫代磷酸酯寡核苷酸保留和分离的影响。为此使用了三种不同的固定相（十八烷基、带有嵌入极性基团的十八烷基和五氟苯基）以及浓度范围为0.1 - 7 mM的离子液体。本研究期间获得的结果表明，离子液体浓度的增加会导致寡核苷酸保留增加。无论使用何种固定相，均观察到这种效应。此外，离子液体结构中烷基链的延长导致反义寡核苷酸保留因子增加。保留研究期间获得的结果证实，向流动相中添加离子液体对反义寡核苷酸保留的影响方式类似于常用离子对试剂（如胺）的情况。还开发了一种寡核苷酸分离方法。对于十八烷基固定相获得了最佳选择性，因为成功分离了序列长度不同的反义寡核苷酸及其代谢物混合物。必须指出的是，离子液体首次用作寡核苷酸分析的流动相添加剂。
