期刊
JOURNAL OF CHROMATOGRAPHIC SCIENCE
卷 58, 期 4, 页码 383-387出版社
OXFORD UNIV PRESS INC
DOI: 10.1093/chromsci/bmz114
关键词
antisense oligonucleotides; hydrophilic interaction; solid phase extraction; silica cartridges
资金
- National Science Centre (Cracow, Poland) under Sonata Bis project [2016/22/E/ST4/00478]
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.
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