Differential metabolomics using stable isotope labeling and two-dimensional gas chromatography with time-of-flight mass spectrometry.

摘要: This work describes an approach to differential metabolomics that involves stable isotope labeling for relative quantification as part of sample analysis by two-dimensional gas chromatography/mass spectrometry (GCxGC/MS). The polar metabolome in control and experimental samples was extracted differentially derivatized using isotopically light heavy (D6) forms the silylation reagent N-methyl-N-tert-butyldimethylsilyl)trifluoroacetamide (MTBSTFA). MTBSTFA derivatives are much greater hydrolytic stability than more common trimethylsilyl derivatives, thus diminishing possibility isotopomer scrambling during GC analysis. Subsequent derivatization with MTBSTFA, labeled were mixed analyzed GCxGC/MS. Metabolites identified, ratio isotopomers quantified. method tested three classes metabolites; amino acids, fatty organic acids. concentration metabolites determined accuracy precision, respectively, standard mixtures 9.5 4.77% 16 9.7 2.83% mixture 19 14 4.53% 20 Suitability examination complex demonstrated analyses spiked blood serum samples. coding proved be effective means compare between two simultaneously.