ASPIRE: Air Pollution: Strategies for Personalized Intervention to Reduce Exposure

The purpose of this subcontract is to fulfill the tasks proposed: To analyze blood plasma of human subjects administered oral 13C6-glucose, for stable isotope resolved polar metabolites depicting glucose metabolism. These tasks include: 1. Perform stable isotope-resolved metabolomic (SIRM) analysis of plasma by rapid extraction 1D 1H and 1H{13C} HSQC NMR for 13C labeled glucose, lactate, Ala, Glu, and other observable 13C labeled metabolites. The results will be used to guide task 2. 2. Perform SIRM analysis of plasma by polar extraction followed by ion-chromatography ultra-high resolution Fourier transform mass spectrometry (IC-UHR-FTMS) [2]. Task 1 results will be used to guide the required volume of plasma, and other parameters, to extract for optimum analysis of 13C labeled metabolites that represent (but not limited to) glycolysis, pentose phosphate pathway (PPP), Krebs cycle, hexosamine phosphate pathway, nucleotide biosynthesis, glutathione biosynthesis, urea cycle and lipid/oxylipin biosynthesis. 3. The data analyses will include, but not limited to, results of the area under curve of the sum of 13C1 plus 13C2 isotopologues of lactate or 13C1 to 13C5 isotopologues of glucose at specified time intervals (e.g. 30, 120 and 180 minutes), expressed as a percentage of the area under curve of total 13C-glucose at these time points as a measure of gluconeogenesis. Additional measures (if observable) include 13C1 plus 13C2 isotopologues of alanine (for gluconeogenesis) and 13C-glutamate (for the Krebs cycle capacity) expressed as percentage of total 13C-glucose.