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anNET affords a generalized framework for Network Embedded Thermodynamic (NET) analysis of quantitative metabolomic datasets. Building upon principles of thermodynamics, this method allows for a quality check of measured metabolite concentrations and enables to spot metabolic reactions where active regulation potentially controls metabolic flux.

The applications of anNET are multiple, for example:

  • Check thermodynamic consistency
  • Estimate unmeasured concentrations
  • Resolve concentrations in different compartments
  • Find minimum/maximum feasible NADH/NAD ratio or adenylate energy charge
  • Infer reaction direction
  • Verify reversibility in model
  • Spot putative control sites
  • Exclude activity of transporters


Matlab 6.0 or higher (tested on Windows and Linux)
Matlab Optimization Toolbox 3 (The Mathworks) or Lindo API (Lindo Systems, Inc.)


  • Kümmel, Panke, and Heinemann, Putative regulatory sites unraveled by network-embedded thermodynamic analysis of metabolome data. Mol Syst Biol. 2006;2:2006.0034. Pubmed
  • Zamboni, Kümmel and Heinemann, anNET: a tool for network-embedded thermodynamic analysis of quantitative metabolome data, BMC Bioinformatics 2008, 9:199. BMC

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