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Methods Enzymol. 2015;560:19-28. doi: 10.1016/bs.mie.2015.03.015. Epub 2015 Jun 2.

Nucleoside Analysis by Hydrophilic Interaction Liquid Chromatography Coupled with Mass Spectrometry.

Author information

1
Department of Chemistry and Biotechnology, Graduate School of Engineering, University of Tokyo, Bunkyo-ku, Tokyo, Japan.
2
Department of Chemistry and Biotechnology, Graduate School of Engineering, University of Tokyo, Bunkyo-ku, Tokyo, Japan. Electronic address: ts@chembio.t.u-tokyo.ac.jp.

Abstract

RNA molecules contain a wide variety of chemical modifications that cannot be deduced from the genomic sequence. RNA modifications confer a chemical diversity to simple RNA molecules, enabling a greater variety of biological functions. To detect RNA modifications, highly sensitive analytical tools are required. Liquid chromatography/mass spectrometry (LC/MS) has been playing a vital role in analyzing minor modified nucleosides in RNA specimens from various sources. Reverse-phase chromatography (RPC) has been used for LC/MS for a long time because RPC is compatible with electrospray ionization (ESI) MS. However, RPC is not always suitable for detecting hydrophilic or polar nucleosides. We here describe a different mode of LC/MS for detecting RNA modifications using hydrophilic interaction liquid chromatography (HILIC). HILIC/ESI-MS is a valuable alternative for profiling modified nucleosides.

KEYWORDS:

HILIC/ESI-MS; Modified nucleoside; Uridine derivative

PMID:
26253964
DOI:
10.1016/bs.mie.2015.03.015
[Indexed for MEDLINE]

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