The application belongs to the field of biological analysis, and provides a method for precise metabolic flow analysis of TCA cycle based on
isotope tracing technology combined with high-
resolution mass spectrometry. By optimizing
mass spectrometry parameters and liquid
chromatography method, one injection within 6 min is realized, and high-
throughput, high-sensitivity and high-resolution detection is realized on 11 biomarkers related to TCA cycle metabolic pathways. At the same time, the application innovatively uses LC-MS / MS to
decipher 13 C the stepwise position-specific transfer from glucose to subsequent metabolites by
glycolysis and TCA cycle, thereby calculating 13 C the integrated steady-state analysis of key metabolic rates of position-specific transfer from sequential precursors to their products (pyruvate
dehydrogenase, beta-oxidation,
pyruvate carboxylase,
isocitrate dehydrogenase and pyruvate cycle). This technology has wide applicability and can potentially characterize mitochondrial
metabolism of any tissue or
cell.