Summary of Study ST001328

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 PR000906. The data can be accessed directly via it's Project DOI: 10.21228/M8KD7Z This work is supported by NIH grant, U2C- DK119886.

See: https://www.metabolomicsworkbench.org/about/howtocite.php

This study contains a large results data set and is not available in the mwTab file. It is only available for download via FTP as data file(s) here.

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Study IDST001328
Study TitleMultiplatform, non-targeted analysis of lung extracts of uninfected and Mtb-infected C57BL/6 mice at 4 and 9 weeks p.i.
Study SummaryThe goal of this multiplatform, non-targeted metabolomics study was to explore the metabolic alterations occurring during the natural progression of pulmonary tuberculosis in a murine model of disease (C57BL/6 genotype). For this purpose, we used gas chromatography, capillary electrophoresis, and reversed-phase liquid chromatography coupled to high-resolution mass analyzers (GC-EI-QTOF/MS, CE-ESI(+)-QTOF/MS, LC-ESI(+)-QTOF/MS and LC-ESI(-)-QTOF/MS to analyze lung extracts of age and sex-matched uninfected mice (UW, n=4), Mycobacterium tuberculosis-infected mice at 4 weeks post-infection (4W, n=4) and Mycobacterium tuberculosis-infected mice at 9 weeks post-infection. All data were acquired in MS1 mode, following a canonical non-targeted workflow.
Institute
Universidad CEU San Pablo
DepartmentDepartamento de Quimica y Bioquimica
LaboratoryCentro de Metabolomica y Bioanalisis (CEMBIO)
Last NameFernandez Garcia
First NameMiguel
AddressFacultad de Farmacia, Universidad San Pablo-CEU, CEU Universities, Urbanización Montepríncipe, 28660 Boadilla del Monte, Spain
Emailmiguel.fernandezgarcia@ceu.es
Phone+34690090778
Submit Date2020-03-12
Num Groups3
Total Subjects13
Num Males6
Num Females7
Raw Data AvailableYes
Raw Data File Type(s)d
Analysis Type DetailGC-MS/LC-MS
Release Date2021-03-15
Release Version1
Miguel Fernandez Garcia Miguel Fernandez Garcia
https://dx.doi.org/10.21228/M8KD7Z
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

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Sample Preparation:

Sampleprep ID:SP001410
Sampleprep Summary:After lung collection, 1 mL of 50 % methanol was added to 100 mg of Mtb-infected or uninfected lung tissue and homogenized in a dounce homogenizer to prepare a uniform suspension. For CE-TOF/MS, 200 uL of homogenate were mixed with 200 L of 0.2 M formic acid and vortexed for 2 min. The samples were cleared by centrifugation at 16000 x g for 10 min at 4 °C and the supernatant was filter-sterilized using 0.22 um spin-X columns (Sigma). For GC-QTOF/MS and LC-QTOF/MS, 200 uL of each sample homogenate were mixed with 800 L of 80:20 methanol:MTBE (Methyl tert-butyl ether) and vortexed for 2 min. Metabolites were then extracted for 1 h with shaking at room temperature and then centrifuged at 4000 x g at 20 °C for 20 min. Supernatants were sterile filtered using 0.22 um Spin-X columns. All samples were passed through a Millipore filter (30-kDa cutoff) to remove large proteins. Samples were dried under high vacuum and stored at -80 °C until further platform-specific processing and analysis. For CE-ESI(+)-TOF/MS analysis, The dried samples were resuspended in Milli-Q water containing 0.1 mM formic acid and 0.2 mM methionine sulfone (internal standard) (Sigma-Aldrich, Germany) by vortexing for 1 min. After subsequent centrifugation (12600 x g, 15 min), the resulting clear solution was analyzed. For GC-QTOF/MS analysis, The above described dried samples were re-suspended in 450 µL of MeOH:H2O:MTBE (74:10:16) and after centrifugation at 12600 x g, 15 min at 4 °C, the supernatant was transferred to a vial with insert and evaporated to dryness under high vacuum. The obtained dried extracts were derivatized by a MPS autosampler for GC/MS analysis as previously described (Fiehn O., 2006). For LC-QTOF/MS analysis, The above described dried samples were resuspended in 200 µL of methanol:water:MTBE (7.4:1:1.6), vortexed for 1.5 h and centrifuged (4000 x g, 10 min, 4 °C). Clear solutions were analyzed.
Sample Derivatization:In GC-EI-QTOF/MS analyses, Briefly, aldehyde and keto groups were first converted to O-methyloximes by reaction with 10 µL pyridine containing 15 mg/mL O-methoxyamine (Sigma-Aldrich, Germany) for 60 min at 70 °C. In a second step, acid hydrogen-containing metabolites were trimethylsilylated by reaction with 10 µL N,O-Bis(trimethylsilyl)trifluoroacetamide (BSTFA) (Sigma-Aldrich, Germany) to enhance the GC/MS metabolite coverage
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