State of the art direct 13C and indirect 1H-[13C] NMR spectroscopy in vivo. A practical guide.

Carbon-13 NMR spectroscopy in combination with (13)C-labeled substrate infusion is a powerful technique for measuring a large number of metabolic fluxes noninvasively in vivo. It has been used to quantify glycogen synthesis rates, establish quantitative relationships between energy metabolism and neurotransmission, and evaluate the importance of different substrates. Measurements can, in principle, be performed through direct (13)C NMR detection or via indirect (1)H-[(13)C] NMR detection of the protons attached to (13)C nuclei. The choice of detection scheme and pulse sequence depends on the magnetic field strength, whereas substrate selection depends on metabolic pathways. (13)C NMR spectroscopy remains a challenging technique that requires several nonstandard hardware modifications, infusion of (13)C-labeled substrates, and sophisticated processing and metabolic modeling. In this study, the various aspects of direct (13)C and indirect (1)H-[(13)C] NMR are reviewed with the aim of providing a practical guide.