Summary of Study ST000019

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 PR000018. The data can be accessed directly via it's Project DOI: 10.21228/M8Z59Q This work is supported by NIH grant, U2C- DK119886.

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Study IDST000019
Study TitleDetermine purity and quality of IROA labelled glucose
Study Type NMR and MS analysis
Study SummaryWe demonstrate the global metabolic analysis ofCaenorhabditis elegansstress responses using a mass-spectrometry-based technique called isotopic ratio outlier analysis (IROA). In an IROA protocol, control and experimental samples are isotopically labeled with 95 and 5%13C, and the two sample populations are mixed together for uniform extraction, sample preparation, and LC-MS analysis.To illustrate the utility of IROA for global metabolomics, we exposed wild-type (N2) worms to a heat shock (30 min heat shock at 33 C), which causes significant, widespread changes in metabolism. We collected and analyzed material from the exometabolome (all material that worms release in the supernatant) and the endometabolome (homogenized total extracts from the worm bodies).
Institute
University of Florida
DepartmentBiochemistry & Molecular Biology
Last NameStupp
First NameGregory
Emailstuppie@ufl.edu
Submit Date2013-12-10
Study CommentsDetermine purity and quality of IROA labelled glucose
Raw Data AvailableYes
Raw Data File Type(s)ser
Uploaded File Size90 M
Analysis Type DetailNMR
Release Date2014-03-05
Release Version1
Gregory Stupp Gregory Stupp
https://dx.doi.org/10.21228/M8Z59Q
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

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Project:

Project ID:PR000018
Project DOI:doi: 10.21228/M8Z59Q
Project Title:Isotopic Ratio Outlier Analysis Global Metabolomics ofCaenorhabditis elegans
Project Type:IROA labeling study
Project Summary:We demonstrate the global metabolic analysis ofCaenorhabditis elegansstress responses using a mass-spectrometry-based technique called isotopic ratio outlier analysis (IROA). In an IROA protocol, control and experimental samples are isotopically labeled with 95 and 5%13C, and the two sample populations are mixed together for uniform extraction, sample preparation, and LC-MS analysis.To illustrate the utility of IROA for global metabolomics, we exposed wild-type (N2) worms to a heat shock (30 min heat shock at 33 C), which causes significant, widespread changes in metabolism. We collected and analyzed material from the exometabolome (all material that worms release in the supernatant) and the endometabolome (homogenized total extracts from the worm bodies).
Institute:University of Florida
Department:Biochemistry & Molecular Biology
Laboratory:Edison
Last Name:Edison
First Name:Arthur
Address:R3-226 Academic Research Building, Department of Biochemistry & Molecular Biology, PO Box 100245, Gainesville, FL 32610-0245
Email:aedison@ufl.edu
Phone:(352) 392-4535

Subject:

Subject ID:SU000036
Subject Type:Animal
Subject Species:-

Factors:

Subject type: Animal; Subject species: - (Factor headings shown in green)

mb_sample_id local_sample_id %13C
SA0011895p0.05
SA00119095p0.95
SA001191Blanknone
Showing results 1 to 3 of 3

Collection:

Collection ID:CO000019
Collection Summary:none
Sample Type:Worms

Treatment:

Treatment ID:TR001190
Treatment Summary:none

Sample Preparation:

Sampleprep ID:SP000032
Sampleprep Protocol Filename:IROAGlucoseNMR_Feb11_2014.txt
Sampleprep Protocol Comments:5%: 10.7 mg dissolved in 594 L of 99% D2O and 6 L of 111.11 mM d4-TSP in D2O 95%: 2.8 mg dissolved in 594 L of 99% D2O and 6 L of 111.11 mM d4-TSP in D2O Blank: 594 L of 99% D2O and 6 L of 111.11 mM d4-TSP in D2O

Combined analysis:

Analysis ID AN000038
Analysis type MS
Chromatography type Reversed phase
Chromatography system Thermo Accela 1250
Column Thermo Scientific Gold aQ
MS Type ESI
MS instrument type Orbitrap
MS instrument name Thermo Q Exactive Orbitrap
Ion Mode POSITIVE
Units

Chromatography:

Chromatography ID:CH000020
Chromatography Comments:100% solvent A for 1 min followed by a linear gradient to 20% B in 6 min, a linear gradient to 60% B in 2 min, a linear gradient to 95% B in 4 min, held for 2 min, and a 3.5 min return to the starting composition
Instrument Name:Thermo Accela 1250
Column Name:Thermo Scientific Gold aQ
Column Temperature:40C
Flow Rate:600 uL/min
Sample Injection:3 uL
Solvent A:0.1% formic acid in water
Solvent B:0.1% formic acid in acetonitrile
Analytical Time:18.5 min
Chromatography Type:Reversed phase

MS:

MS ID:MS000035
Analysis ID:AN000038
Instrument Name:Thermo Q Exactive Orbitrap
Instrument Type:Orbitrap
MS Type:ESI
MS Comments:LCMSmethod.rtf
Ion Mode:POSITIVE
Mass Accuracy:1.5 ppm
Spray Voltage:3kV
Resolution Setting:70000
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