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Breath water-based doubly labelled water method for the noninvasive determination of CO 2 production and energy expenditure in mice.

We explored a novel doubly labelled water (DLW) method based on breath water (BW-DLW) in mice to determine whole body CO2 production and energy expenditure noninvasively. The BW-DLW method was compared to the DLW based on blood plasma. Mice (n = 11, 43.5 ± 4.6 g body mass (BM)) were administered orally a single bolus of doubly labelled water (1.2 g H2 18 O kg BM-1 and 0.4 g 2 H2 O kg BM-1 , 99 atom% (AP) 18 O or 2 H). To sample breath water, the mice were placed into a respiration vessel. The exhaled water vapour was condensed in a cold-trap. The isotope enrichments of breath water were compared with plasma samples. The 2 H/1 H and 18 O/16 O isotope ratios were measured by means of isotope ratio mass spectrometry. The CO2 production (RCO2 ) was calculated from the 2 H and 18 O enrichments in breath water and plasma over 5 days. The isotope enrichments of breath water vs. plasma were correlated (R2  = 0.89 for 2 H and 0.95 for 18 O) linearly. The RCO2 determined based on breath water and plasma was not different (113.2 ± 12.7 vs. 111.4 ± 11.0 mmol d-1 ), respectively. In conclusion, the novel BW-DLW method is appropriate to obtain reliable estimates of RCO2 avoiding blood sampling.

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