Com (Zea mays L.) root tips were subjected to different conditions so that nucleotide levels varied over a wide range. Levels of nucleotides in com root tips were measured using 31P nuclear magnetic resonance (NMR) spectroscopy and high performance liquid chromatography. Compartmentation of nucleotides in cells can also be inferred from metabolite analysis of near-equilibrium reactions.The mass action ratio for a reaction known to operate near equilibrium in vivo is determined by analysis of the whole cell or tissue. Compartmentation of one or more substrates away from the particular enzyme is suggested when the mass action ratio differs significantly from the equilibrium constant. Using this approach, together with a comparison of tissues differing in mitochondrial content, Veech et al. (29) concluded that much of the ADP in cells is sequestered in mitochondria, and therefore inaccessible to cytoplasmic enzymes on the timescale of nucleotide turnover (seconds