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9%. 01% (104 dpm/µmol). It is easy to see that 13C meets neither the criteria for low background signal or that for a "massless"tracer. However, it is not always necessary to provide very large fractional enrichments for the sake of detection. Ifthe tissue can be excised and extracted, the labeled molecules are stable and 13C NMR spectra can be taken over many hours at much higher fields (yielding higher resolution and S/N) than are generally available for in situ experiments. A lower limit for the fractional enrichment of the substrate in this instance is set not by the ability to detect label, but by the error in detecting 13C over the natural abundance.

These two molecules have the same splitting patterns in C4 and C3, but in the first C2 will be a quartet, and in the second, a doublet. It is clear that even though the only new bond that is ever made is that between C2 of oxaloacetate and C2 of acetyl-CoA, the splitting patterns become increasingly more complex with more turns of the cycle. The patterns can therefore be used to determine the relative length of time (or number of turns) that each molecule remains in the TCA cycle. If the labeled substrate can enter the cycle through an anaplerotic pathway, such as pyruvate carboxylase, the isotopic pattern will be altered again in a predictable way.

1993) demonstrated a large gradient in creatine kinase flux across the wall of the left ventricle. It has also been shown that the PCr/ATP ratio is elevated in vivo in the right ventricle muscle, but not the left ventricle, during catecholamine infusion. , 1989). Most other NMR accessible organs are even more heterogeneous. , 1989). , 1991). The model development process should therefore consider the question: How will the validity of the model be affected by heterogeneity in this tissue? LaughlinandJoanne K.

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