Application of ion mobility-mass spectrometry to microRNA analysis

Kosuke Takebayashi, Kenji Hirose, Yoshihiro Izumi, Takeshi Bamba, Eiichiro Fukusaki

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

Liquid chromatography/mass spectrometry is widely used for studying sequence determination and modification analysis of small RNAs. However, the efficiency of liquid chromatography-based separation of intact small RNA species is insufficient, since the physiochemical properties among small RNAs are very similar. In this study, we focused on ion mobility-mass spectrometry (IM-MS), which is a gas-phase separation technique coupled with mass spectrometry; we have evaluated the utility of IM-MS for microRNA (miRNA) analysis. A multiply charged deprotonated ion derived from an 18-24-nt-long miRNA was formed by electrospray ionization, and then the time, called the " drift time" , taken by each ion to migrate through a buffer gas was measured. Each multivalent ion was temporally separated on the basis of the charge state and structural formation; 3 types of unique mass-mobility correlation patterns (i.e., chainlike-form, hairpin-form, and dimer-form) were present on the two-dimensional mobility-mass spectrum. Moreover, we found that the ion size (sequence length) and the secondary structures of the small RNAs strongly contributed to the IM-MS-based separation, although solvent conditions such as pH had no effect. Therefore, sequence isomers could also be discerned by the selection of each specific charged ion, i.e., the 6- charged ion reflected a majority among chainlike-, hairpin-, and other structures. We concluded that the IM-MS provides additional capability for separation; thus, this analytical method will be a powerful tool for comprehensive small RNA analysis.

Original languageEnglish
Pages (from-to)332-338
Number of pages7
JournalJournal of Bioscience and Bioengineering
Volume115
Issue number3
DOIs
Publication statusPublished - Mar 1 2013
Externally publishedYes

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MicroRNAs
Mass spectrometry
Mass Spectrometry
Ions
RNA
Liquid chromatography
Liquid Chromatography
Gases
Electrospray ionization
Phase separation
Isomers
Dimers
Sequence Analysis
Buffers

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology

Cite this

Application of ion mobility-mass spectrometry to microRNA analysis. / Takebayashi, Kosuke; Hirose, Kenji; Izumi, Yoshihiro; Bamba, Takeshi; Fukusaki, Eiichiro.

In: Journal of Bioscience and Bioengineering, Vol. 115, No. 3, 01.03.2013, p. 332-338.

Research output: Contribution to journalArticle

Takebayashi, Kosuke ; Hirose, Kenji ; Izumi, Yoshihiro ; Bamba, Takeshi ; Fukusaki, Eiichiro. / Application of ion mobility-mass spectrometry to microRNA analysis. In: Journal of Bioscience and Bioengineering. 2013 ; Vol. 115, No. 3. pp. 332-338.
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