BB-rel@ldeleger:BB-rel-18694716 / 96-1409 JSONTXT

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bionlp-ost-19-BB-rel-dev

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Id Subject Object Predicate Lexical cue
T2 0-1313 Paragraph denotes In this report, we introduce a liquid chromatography single-mass spectrometry method for metabolome quantification, using the LTQ Orbitrap high-resolution mass spectrometer. Analytes were separated with hydrophilic interaction liquid chromatography. At a working resolution of 30,000 (at m/z 400), the limit of detection varied from 50 fmol to 5 pmol for 25 metabolites tested. In terms of metabolite concentration, the linearity was about 2 to 3 orders of magnitude for most compounds (R(2)>0.99). To determine the accuracy of the system in complex sample matrices, the isotope dilution method was evaluated from mixtures of pure compounds and uniformly 13C-labeled cell extracts. With the application of this method, quantification was possible within single runs even when the pool sizes of individual metabolites varied from 0.13 to 55.6 microM. As a case study, intracellular concentrations of central metabolites were determined for Methylobacterium extorquens AM1 during growth on two different carbon sources, methanol and succinate. Reproducible results from technical and biological repetitions were obtained that revealed significant variations of intracellular metabolite pool sizes, depending on the carbon source. The LTQ Obitrap offers new perspectives and strategies for metabolome quantification.
T3 126-172 Habitat denotes LTQ Orbitrap high-resolution mass spectrometer
T4 614-680 Habitat denotes mixtures of pure compounds and uniformly 13C-labeled cell extracts
T5 655-671 Habitat denotes 13C-labeled cell
T6 939-970 Microorganism denotes Methylobacterium extorquens AM1
T7 1232-1243 Habitat denotes LTQ Obitrap