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Methods and Instrumentation

 

I. LC-ESI-MS Coupling for Detection, Structure Elucidation and Quantification of Metabolites

 

1. LC-ESI-QqQ Mass Spectrometry (Triple Quadrupol MS)

LC-QqQ

 
  • quantification of metabolites
  • systematic search for structural homologues
  • elucidation of metabolic pathways
  • detection of minor metabolites in complex matrices
  • semiquantitative determination of substance classes
  • verification of labelling experiments
  • flux analysis

 

2. LC Ion Trap Mass Spectrometry (IT-MS)

 
  • determination of fragmentation hierarchies: structure elucidation of metabolites and degradation products
  • systematic search for structural homologues
  • multiplexing of substance classes
  • flux analysis
  • identification of modifications

3. LC-QqTOF Mass Spectrometry (ESI-TOF-MS)

 
  • metabolic profiling: exact mass database search (Metlin database)
  • structure analysis of unknown compounds
  • elucidation of metabolic pathways
  • identification of modifications

4. Isotope Ratio Mass Spectrometry (IR-MS)

  • isotope ratios of C, N, O
  • flux analysis
  • elemental analysis of C, N, O

 

 

II. GC-EI-MS Coupling for Metabolic Profiling in plants, animals and microorganisms

 

gc_system.JPG

As a routine method we are profiling relative concentration changes in primary metabolites (sugars, sugar-alcohols, amino acids, organic acids and phosphorylated compounds). The mainly polar metabolites are extracted from the tissue, derivatised and the derivates are quantified and identified with an Agilent GC-Quadrupole-MS System, which can operate in EI and CI mode.

Several more targeted methods for analysing particular (also non-polar) compounds are developed in cooperation with the various users.

 

 

GC-MS chromatogram

Fig.1: Example Chromatogram for a GC-MS primary metabolite profiling in Physcomitrella patens

 

 

cluster analysis

Fig.2: Example for a cluster analysis for the relative changes of metabolite concentrations in Ricinus communis under anoxia.