Back to Search
Start Over
Targeted Determination of Tissue Energy Status by LC-MS/MS
- Source :
- Analytical Chemistry. 91:5881-5887
- Publication Year :
- 2019
- Publisher :
- American Chemical Society (ACS), 2019.
-
Abstract
- [Image: see text] Intracellular nucleotides and acyl-CoAs are metabolites that are central to the regulation of energy metabolism. They set the 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, and ATP), pyridine dinucleotides (NAD(+) and 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.
- Subjects :
- chemistry.chemical_classification
Nucleotides
010401 analytical chemistry
Allosteric regulation
010402 general chemistry
Tandem mass spectrometry
01 natural sciences
0104 chemical sciences
Analytical Chemistry
Mice
chemistry.chemical_compound
Liver
Biosynthesis
chemistry
Tandem Mass Spectrometry
Adenine nucleotide
Biophysics
Animals
Nucleotide
Acyl Coenzyme A
NAD+ kinase
Energy charge
Quantitative analysis (chemistry)
Chromatography, Liquid
Subjects
Details
- ISSN :
- 15206882 and 00032700
- Volume :
- 91
- Database :
- OpenAIRE
- Journal :
- Analytical Chemistry
- Accession number :
- edsair.doi.dedup.....1f94c2b61b1584d283f31930fbf96b7e
- Full Text :
- https://doi.org/10.1021/acs.analchem.9b00217