title: Innovations in generation and analysis of 2D [(13)C,(1)H] COSY NMR spectra for metabolic flux analysis purposes.
abstract: 2D [(13)C,(1)H] COSY NMR is used by the metabolic engineering community for determining (13)C-(13)C connectivities in intracellular compounds that contain information regarding the steady-state fluxes in cellular metabolism. This paper proposes innovations in the generation and analysis of these specific NMR spectra. These include a computer tool that allows accurate determination of the relative peak areas and their complete covariance matrices even in very complex spectra. Additionally, a method is introduced for correcting the results for isotopic non-steady-state conditions. The proposed methods were applied to measured 2D [(13)C,(1)H] COSY NMR spectra. Peak intensities in a one-dimensional section of the spectrum are frequently not representative for relative peak volumes in the two-dimensional spectrum. It is shown that for some spectra a significant amount of additional information can be gained from long-range (13)C-(13)C scalar couplings in 2D [(13)C,(1)H] COSY NMR spectra. Finally, the NMR resolution enhancement by dissolving amino acid derivatives in a nonpolar solvent is demonstrated.
authors.affiliation: Bioprocestechnology Group, Faculty of Applied Sciences, Delft University of Technology, Julianalaan 67, 2628 BC Delft, The Netherlands. W.A.VanWinden@tnw.tudelft.nl
authors.forename: W
authors.initials: W
authors.lastname: van Winden
authors.forename: D
authors.initials: D
authors.lastname: Schipper
authors.forename: P
authors.initials: P
authors.lastname: Verheijen
authors.forename: J
authors.initials: J
authors.lastname: Heijnen
mesh_terms: D000596:Amino Acids / Q000378:metabolism*; D002247:Carbon Isotopes; D009682:Magnetic Resonance Spectroscopy
chemical_list: D000596:Amino Acids; D002247:Carbon Isotopes