13C Protein Oxidation in Breath: Is It Relevant for the Whole Body Protein Status?

Introduction: The kinetics of protein oxidation, monitored in breath, and its contribution to the whole body protein status is not well established. Objectives: To analyze protein oxidation in various metabolic conditions we developed/validated a 13C-protein oxidation breath test using low enriched milk proteins. Method/Design: 30 g of naturally labeled 13C-milk proteins were consumed by young healthy volunteers. Breath samples were taken every 10 min and 13CO2 was measured by Isotope Ratio Mass Spectrometry. To calculate the amount of oxidized substrate we used: substrate dose, molecular weight and 13C enrichment of the substrate, number of carbon atoms in a substrate molecule, and estimated CO2-production of the subject based on body surface area. Results: We demonstrated that in 255 min 20% ± 3% (mean ± SD) of the milk protein was oxidized compared to 18% ± 1% of 30 g glucose. Postprandial kinetics of oxidation of whey (rapidly digestible protein) and casein (slowly digestible protein) derived from our breath test were comparable to literature data regarding the kinetics of appearance of amino acids in blood. Oxidation of milk proteins was faster than that of milk lipids (peak oxidation 120 and 290 minutes, respectively). After a 3-day protein restricted diet (~10 g of protein/day) a decrease of 31% ± 18% in milk protein oxidation was observed compared to a normal diet. Conclusions: Protein oxidation, which can be easily monitored in breath, is a significant factor in protein metabolism. With our technique we are able to characterize changes in overall protein oxidation under various metabolic conditions such as a protein restricted diet, which could be relevant for defining optimal protein intake under various conditions. Measuring protein oxidation in new-born might be relevant to establish its contribution to the protein status and its age-dependent development.