Spectral trees as a robust annotation tool in LC–MS based metabolomics

Research output: Contribution to journalArticleAcademicpeer-review

57 Citations (Scopus)

Abstract

The identification of large series of metabolites detectable by mass spectrometry (MS) in crude extracts is a challenging task. In order to test and apply the so-called multistage mass spectrometry (MS n ) spectral tree approach as tool in metabolite identification in complex sample extracts, we firstly performed liquid chromatography (LC) with online electrospray ionization (ESI)–MS n , using crude extracts from both tomato fruit and Arabidopsis leaf. Secondly, the extracts were automatically fractionated by a NanoMate LC-fraction collector/injection robot (Advion) and selected LC-fractions were subsequently analyzed using nanospray-direct infusion to generate offline in-depth MS n spectral trees at high mass resolution. Characterization and subsequent annotation of metabolites was achieved by detailed analysis of the MS n spectral trees, thereby focusing on two major plant secondary metabolite classes: phenolics and glucosinolates. Following this approach, we were able to discriminate all selected flavonoid glycosides, based on their unique MS n fragmentation patterns in either negative or positive ionization mode. As a proof of principle, we report here 127 annotated metabolites in the tomato and Arabidopsis extracts, including 21 novel metabolites. Our results indicate that online LC–MS n fragmentation in combination with databases of in-depth spectral trees generated offline can provide a fast and reliable characterization and annotation of metabolites present in complex crude extracts such as those from plants.
Original languageEnglish
Pages (from-to)691-703
JournalMetabolomics
Volume8
Issue number4
DOIs
Publication statusPublished - 2012

Keywords

  • orbitrap mass-spectrometry
  • liquid-chromatography
  • tomato fruit
  • secondary metabolites
  • phenolic-compounds
  • brassica-rapa
  • hplc-dad
  • identification
  • flavonoids
  • arabidopsis

Fingerprint Dive into the research topics of 'Spectral trees as a robust annotation tool in LC–MS based metabolomics'. Together they form a unique fingerprint.

Cite this