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Targeted Determination of Tissue Energy Status by LC-MS/MS.

08:00 EDT 2nd April 2019 | BioPortfolio

Summary of "Targeted Determination of Tissue Energy Status by LC-MS/MS."

Intracellular nucleotides and acyl-CoAs are metabolites that are central to the regulation of energy metabolism. They set cellular energy charge and redox state, act as allosteric regulators, modulate signaling and transcription factors, and thermodynamically activate substrates for oxidation or biosynthesis. Unfortunately, no method exists to simultaneously quantify these biomolecules in tissue extracts. A simple method was developed using ion-pairing reversed phase high performance liquid chromatography-electrospray ionization tandem mass spectrometry (HPLC-ESI-MS/MS) to simultaneously quantify adenine nucleotides (AMP, ADP, ATP), pyridine dinucleotides (NAD+, NADH) and short-chain acyl-CoAs (acetyl, malonyl, succinyl and propionyl). Quantitative analysis of these molecules in mouse liver was achieved using stable isotope labeled internal standards. The method was extensively validated by determining the linearity, accuracy, repeatability, and assay stability. Biological responsiveness was confirmed in assays of liver tissue with variable durations of ischemia, which had substantial effects on tissue energy charge and redox state. We conclude that the method provides a simple, fast and reliable approach to the simultaneous analysis of nucleotides and short-chain acyl-CoAs.

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This article was published in the following journal.

Name: Analytical chemistry
ISSN: 1520-6882
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