.sup.13 C Isotopomer analyses in intact tissue using (.sup.13 C)

Drug – bio-affecting and body treating compositions – In vivo diagnosis or in vivo testing – Magnetic imaging agent

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435 35, 435 4, 435 14, 435 29, 435 30, 436 59, 436 57, 436 63, 436173, 424 935, 424 181, 1286534, 128654, A61K 5104, C12Q 100, C12Q 102, C12Q 104

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055975488

ABSTRACT:
Entry of .sup.13 C-enriched acetyl-CoA into the citric acid cycle results in scrambling of .sup.13 C into the various carbon positions of all intermediate pools. The eventual result is that the .sup.13 C resonances of all detectable intermediates or molecules exchanging with those intermediates appear as multiplets due to nearest neighbor spin-spin couplings. Isotopomer analysis of the glutamate .sup.13 C multiplets provides a history of .sup.13 C flow through the cycle pools. Relative substrate utilization and relative anaplerotic flux can be quantitated. A major limitation of the method for in vivo applications is spectral resolution of multi-line resonances required for a complete isotopomer analysis. It is now shown that (.sup.13 C)homonuclear decoupling of the glutamate C3 resonance collapses nine-line C4 and C2 resonances into three line multiplets. These three-line .sup.13 C multiplets are well resolved in isolated, perfused rat hearts and present steady-state equations which allow an isotopomer analysis from data obtained in intact tissue. This advancement shows for the first time that .sup.13 C isotopomer methods may be extended to complex metabolic conditions for resolution of carbon-carbon coupling, and particularly to in vivo measurements.

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