Product Details of 329-89-5. The protonation of heteroatoms in aromatic heterocycles can be divided into two categories: lone pairs of electrons are in the aromatic ring conjugated system; and lone pairs of electrons do not participate. Compound: 6-Aminonicotinamide, is researched, Molecular C6H7N3O, CAS is 329-89-5, about Ex vivo and in vivo stable isotope labelling of central carbon metabolism and related pathways with analysis by LC-MS/MS. Author is Yuan, Min; Kremer, Daniel M.; Huang, He; Breitkopf, Susanne B.; Ben-Sahra, Issam; Manning, Brendan D.; Lyssiotis, Costas A.; Asara, John M..
Targeted tandem mass spectrometry (LC-MS/MS) has been extremely useful for profiling small mols. extracted from biol. sources, such as cells, bodily fluids and tissues. Here, we present a protocol for analyzing incorporation of the non-radioactive stable isotopes carbon-13 (13C) and nitrogen-15 (15N) into polar metabolites in central carbon metabolism and related pathways. Our platform utilizes selected reaction monitoring (SRM) with polarity switching and amide hydrophilic interaction liquid chromatog. (HILIC) to capture transitions for carbon and nitrogen incorporation into selected metabolites using a hybrid triple quadrupole (QQQ) mass spectrometer. This protocol represents an extension of a previously published protocol for targeted metabolomics of unlabeled species and has been used extensively in tracing the metabolism of nutrients such as 13C-labeled glucose, 13C-glutamine and 15N-glutamine in a variety of biol. settings (e.g., cell culture experiments and in vivo mouse labeling via i.p. injection). SRM signals are integrated to produce an array of peak areas for each labeling form that serve as the output for further anal. The processed data are then used to obtain the degree and distribution of labeling of the targeted mols. (termed fluxomics). Each method can be customized on the basis of known unlabeled Q1/Q3 SRM transitions and adjusted to account for the corresponding 13C or 15N incorporation. The entire procedure takes ∼6-7 h for a single sample from exptl. labeling and metabolite extraction to peak integration.
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