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. 2019 Apr 2;91(9):5881–5887. doi: 10.1021/acs.analchem.9b00217

Figure 1.

Figure 1

(A) General roles of adenosine phosphates, dinucleotides, and short-chain acyl-CoAs in metabolism and the energy status of liver. Nutrient oxidation in pathways like glycolysis and fat oxidation captures electrons by reducing NAD+ to NADH. These pathways also generate acetyl-CoA, which is oxidized in the TCA cycle to generate more NADH. NADH is reoxidized to NAD+ in the respiratory chain to drive ATP synthesis. The conversion of ATP to ADP and AMP supplies the free energy for multiple pathways of cellular work. Many of these metabolites can be sensed by regulatory proteins, such as AMPK and sirtuins, or allosterically modify enzymatic activity to signal for changes in flux through pathways of nutrient oxidation and biosynthesis. (B) Analytical methodology designed for the simultaneous quantification of adenosine phosphates, dinucleotides, and short-chain acyl-CoAs in tissue samples. Snap-freezing normoxic tissue ensures that physiological pools of metabolites are preserved. Acid extraction provides sufficient extraction efficiency for these classes of metabolites. Ion-pairing with DBAA provides sufficient chromatographic separation of these classes of metabolites. Positive MS/MS fragments were more efficient than negative-mode ones under these conditions.