Summary of Study ST001608
This data is available at the NIH Common Fund's National Metabolomics Data Repository (NMDR) website, the Metabolomics Workbench, https://www.metabolomicsworkbench.org, where it has been assigned Project ID PR001033. The data can be accessed directly via it's Project DOI: 10.21228/M85M45 This work is supported by NIH grant, U2C- DK119886.
See: https://www.metabolomicsworkbench.org/about/howtocite.php
Study ID | ST001608 |
Study Title | Comparing gas chromatography with time-of-flight, quadrupole time-of-flight and quadrupole mass spectrometry for stable isotope tracing (part-I) |
Study Summary | Stable isotope tracers are applied in vivo and in vitro studies to reveal the activity of enzymes and intracellular metabolic pathways. Most often, such tracers are used with gas chromatography coupled to mass spectrometry (GC-MS) due to its ease of operation and reproducible mass spectral databases. Differences in isotope tracer performance of classic GC-quadrupole MS instrument and newer time-of-flight instruments are not well-studied. Here, we used three commercially available instruments for the analysis of identical samples from a stable isotope labeling study that used [U-13C6] d-glucose to investigate the metabolism of Rothia mucilaginosa with respect to 29 amino acids and hydroxyl acids involved in primary metabolism. Overall, all three GC-MS instruments (low-resolution GC-SQ-MS, low-resolution GC-TOF-MS, and high-resolution GC-Q-TOF-MS) can be used to perform stable isotope tracing studies for glycolytic intermediates, TCA metabolites and amino acids, yielding similar biological results, with high-resolution GC-Q-TOF-MS offering additional capabilities to identify chemical structures of unknown compounds that might show significant isotope enrichments in biological studies. |
Institute | University of California, Davis |
Laboratory | Oliver Fiehn |
Last Name | Zhang |
First Name | Ying |
Address | West Coast Metabolomics Center, University of California, Davis, 95616, CA, USA |
ythzhang@ucdavis.edu | |
Phone | +1-530-754-8258 |
Submit Date | 2020-11-19 |
Raw Data Available | Yes |
Raw Data File Type(s) | d |
Analysis Type Detail | GC-MS |
Release Date | 2021-05-19 |
Release Version | 1 |
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Collection:
Collection ID: | CO001678 |
Collection Summary: | A quality control mixture of 29 unlabeled metabolite standards (Supporting information Table SI 1) was prepared to reach a final concentration of 1 mg/mL as stock solution. 50 µL aliquot from each stock solution was combined into a new tube and dried down. Then, the mixture was diluted to reach a final concentration of 50 µg/mL working solution. Rothia mucilaginosa strain RmFLR01 was isolated from a cystic fibrosis (CF) patient at the UC San Diego Adult CF Clinic. R. mucilaginosa cultures were grown in triplicates in artificial-sputum medium spiked with 100 mM [U-13C6] d-glucose (Cambridge Isotope Laboratory, Tewksbury, MA, USA) under anaerobic and aerobic conditions (5% CO2) at 37°C and harvested at 4, 8, 12 and 24 h for isotope tracer analyses. |
Sample Type: | Bacterial cells |