A novel stable isotope labelling assisted workflow from improved untargeted LC-HRMS based metabolomics research

Adriano Marocco, Christoph Bueschl, Bernhard Kluger, Marc Lemmens, Gerhard Adam, Gerlinde Wiesenberger, Joseph Strauss, Stephan Bödi, Gerhard G. Thallinger, Rudolf Krska, Rainer Schuhmacher

Risultato della ricerca: Contributo in rivistaArticolo in rivistapeer review

47 Citazioni (Scopus)

Abstract

Many untargeted LC–ESI–HRMS based metabolomics studies are still hampered by the large proportion of non-biological sample derived signals included in the generated raw data. Here, a novel, powerful stable isotope labelling (SIL)-based metabolomics workflow is presented, which facilitates global metabolome extraction, improved metabolite annotation and metabolome wide internal standardisation (IS). The general concept is exemplified with two different cultivation variants, (1) co-cultivation of the plant pathogenic fungi Fusarium graminearum on nonlabelled and highly 13C enriched culture medium and (2) experimental cultivation under native conditions and use of globally U-13C labelled biological reference samples as exemplified with maize and wheat. Subsequent to LC– HRMS analysis of mixtures of labelled and non-labelled samples, two-dimensional data filtering of SIL specific isotopic patterns is performed to better extract truly biological derived signals together with the corresponding number of carbon atoms of each metabolite ion. Finally, feature pairs are convoluted to feature groups each representing a single metabolite. Moreover, the correction of unequal matrix effects in different sample types and the improvement of relative metabolite quantification with metabolome wide IS are demonstrated for the F. graminearum experiment. Data processing employing the presented workflow revealed about 300 SIL derived feature pairs corresponding to 87–135 metabolites in F. graminearum samples and around 800 feature pairs corresponding to roughly 350 metabolites in wheat samples. SIL assisted IS, by the use of globally U-13C labelled biological samples, reduced the median CV value from 7.1 to 3.6 % for technical replicates and from 15.1 to 10.8 % for biological replicates in the respective F. graminearum samples.
Lingua originaleEnglish
pagine (da-a)754-769
Numero di pagine16
RivistaMetabolomics
Volume10
DOI
Stato di pubblicazionePubblicato - 2014

Keywords

  • 13C.labelling
  • Fusarium
  • Internal standardization
  • Maize
  • Metabolomics
  • Wheat

Fingerprint Entra nei temi di ricerca di 'A novel stable isotope labelling assisted workflow from improved untargeted LC-HRMS based metabolomics research'. Insieme formano una fingerprint unica.

Cita questo