Nuckowski Łukasz, Kilanowska Anna, Studzińska Sylwia
Chair of Environmental Chemistry and Bioanalytics, Faculty of Chemistry, Nicolaus Copernicus University in Toruń, 7 Gagarin St., PL-87-100 Toruń, Poland.
J Chromatogr Sci. 2020 Apr 23;58(4):383-387. doi: 10.1093/chromsci/bmz114.
The presented studies aimed to develop a new and simple extraction method based on hydrophilic interaction for antisense oligonucleotides with different modifications. For this purpose, solid-phase extraction cartridges with unmodified silica were used. All extraction steps were performed by utilizing water, acetonitrile, acetone or their mixtures. The results obtained show that a high content (95%) of organic solvent, used during sample loading, is critical to achieve a successful extraction, while elution with pure water allows effective oligonucleotides desorption. The recovery values were greater than 90% in the case of unmodified DNA, phosphorothioate, 2'-O-(2-methoxyethyl) and 2'-O-methyl oligonucleotides. For the mixture of phosphorothioate oligonucleotide and its two synthetic metabolites, the recovery values for the standard solutions were in the range of 70-75%, while for spiked human plasma, 45-50%. The developed method is simple, may be performed in a short time and requires simple solvents like water or acetonitrile/acetone, thus showing promise as an alternative to chaotropic salt-based or ion pair-based SPE methods.
所展示的研究旨在开发一种基于亲水相互作用的新型简单提取方法，用于提取具有不同修饰的反义寡核苷酸。为此，使用了未修饰硅胶的固相萃取柱。所有提取步骤均通过使用水、乙腈、丙酮或它们的混合物来进行。所得结果表明，在样品加载过程中使用高含量（95%）的有机溶剂对于实现成功提取至关重要，而用纯水洗脱可实现有效的寡核苷酸解吸。对于未修饰的DNA、硫代磷酸酯、2'-O-(2-甲氧基乙基)和2'-O-甲基寡核苷酸，回收率大于90%。对于硫代磷酸酯寡核苷酸及其两种合成代谢物的混合物，标准溶液的回收率在70 - 75%范围内，而对于加标的人血浆，回收率为45 - 50%。所开发的方法简单，可在短时间内完成，并且需要水或乙腈/丙酮等简单溶剂，因此有望成为基于离液盐或离子对的固相萃取方法的替代方法。
