Post-mortem energy metabolism in bovine muscles studied by non-invasive phosphorus-31 nuclear magnetic resonance

Abstract

Phosphorus-31 Nuclear Magnetic Resonance ((31)P-NMR) has been utilized to follow non-invasively the post-mortem metabolism of the major phosphorylated metabolites in muscles from beef slaughter carcasses. In addition to adenosine-5'-triphosphate (ATP), creatine phosphate (CP) and inorganic phosphate (P(i)) considerable amounts of glucose- and fructose-6-phosphate (G6P and F6P, respectively) as well as glycerol-3-phosphate (Glyc3P) were detected. ATP was mainly present as a Mg(2+)-ATP complex. Adenosine-5'-diphosphate (ADP) appeared to be mainly bound to muscle proteins. A good quantitative agreement was found for the levels of ATP, CP and sugar phosphates (SP) when estimated by NMR or enzymatic assays. Since the chemical shifts of the P(i) and sugar phosphate resonances are a function of the pH, the intracellular pH could be directly deduced from the NMR spectra. Values obtained in this manner were, within the errors of both methods, the same as those determined in iodoacetate/KCl homogenates. The pH gradients within the tissue never exceeded 0.3 pH units. In a final set of experiments we used (31)P-NMR 10 study the effects of electrical stimulation on the intracellular pH and post-mortem metabolism. It was concluded that (31)P-NMR, due to its non-invasive nature plus the fact that some of the NMR parameters are sensitive to the intracellular environment, provides a useful complement to existing methods for the study of post-mortem metabolism.