Two methods of measuring rates of gluconeogenesis based on label redistribution after the introduction of [U-(13)C]glucose into the whole body are examined. These methods are compared with methods previously derived for carbon-14 tracers. It is shown that the three approaches (stoichiometric, dilution, and combinatorial) are equivalent, provided the same set of assumptions are used. Barring a factor of two [see Am. J. Physiol. 270 (Endocrinol. Metab. 33): E709-E717, 1996], the differences ( approximately 10-15%) in the carbon-based dilutional and the molecule-based estimates of the rate of gluconeogenesis from published isotopomer data likely arise from small differences in the assumptions that concern the relative rate of label loss from the different isotopomers. The production of unlabeled substrate for glucose synthesis (phosphoenolpyruvate) from the different isotopomers of lactate is shown to be a potential source of error in these methods. This error is estimated using models of the interaction of the gluconeogenetic pathway and the tricarboxylic acid (TCA) cycle and is shown to vary from negligible to 30% depending on the relative flux of the two pathways through the oxaloacetate pool. Because the estimates obtained by both methods considered are lower than is physiologically expected, some of the assumptions made may not hold. Future work will exploit the rich information content of isotopomer data to yield improved estimates.