High-Speed Tracer Analysis of Metabolism (HS-TrAM)

Tracing the fate of stable isotopically-enriched nutrients is a sophisticated
method of describing and quantifying the activity of metabolic pathways.
Nuclear Magnetic Resonance (NMR) offers high resolution data, yet is
under-utilised due to length of time required to collect the data, quantification
requiring multiple samples and complicated analysis. Here we present two
techniques, quantitative spectral filters and enhancement of the splitting due to
J-coupling in H, C-HSQC NMR spectra, which allow the rapid collection of
NMR data in a quantitative manner on a single sample. The reduced duration of
HSQC spectra data acquisition opens up the possibility of real-time tracing of
metabolism including the study of metabolic pathways in vivo. We show how
these novel techniques can be used to trace the fate of labelled nutrients in a
whole organ model of kidney preservation prior to transplantation using a
porcine kidney as a model organ, and also show how the use of multiple
nutrients, differentially labelled with C and N, can be used to provide
additional information with which to profile metabolic pathways.