Changes in cerebral macromolecular 1H nuclear magnetic resonance (NMR) spectrum were studied in cortical brain slices in vitro. Aglycaemic hypoxia irreversibly increased various short T2 spectral components at 1.8-0.8 ppm in concordance with energy loss and independent of T1 and T2 relaxation effects. Removal of external calcium (Ca2+e) slightly attenuated the effect. The results suggest NMR-visible reorganisation of intracellular proteins due to hypoxic insult, and show that it may be possible to monitor early cytoplasmic changes due to brain energy depletion by NMR spectroscopy.