Summary of study ST000025

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 PR000024. The data can be accessed directly via it's Project DOI: 10.21228/M85P4V 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 IDST000025
Study TitleMetabolomics Analysis of Thermally Challenged Mayfly Larvae (GCMS analysis)
Study TypeMetabolomic analysis of mayflies
Study SummaryThe purpose of this study was to examine the metabolic profiles of mayfly (Centroptilum triangulifer) larvae subjected to thermal challenge. This species is unusual in terms of its ease of culture, and its suitability as a laboratory test organism. Our purpose here was to examine how an environmentally realistic thermal challenge affects the physiology of this organism. In this study, we obtained several types of insect species and we were able to show that GC-MS Metabolomics could be used to distinguish among the different types of larvae.
Institute
University of North Carolina
DepartmentSystems and Translational Sciences
LaboratorySumner Lab
Last NameSumner
First NameSusan
AddressEastern Regional Comprehensive Metabolomics Resource Core, UNC Nutrition Research Institute, 500 Laureate Way, Kannapolis, NC, 28081
Emailsusan_sumner @unc.edu
Phone704-250-5066
Submit Date2014-02-28
Num Groups7
Total Subjects13
Raw Data AvailableYes
Raw Data File Type(s).cdf files
Uploaded File Size870 M
Analysis Type DetailGC-MS
Release Date2015-03-12
Release Version1
Susan Sumner Susan Sumner
https://dx.doi.org/10.21228/M85P4V
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

Select appropriate tab below to view additional metadata details:


Project:

Project ID:PR000024
Project DOI:doi: 10.21228/M85P4V
Project Title:Metabolomics Analysis of Thermally Challenged Mayfly Larvae
Project Type:MS/NMR
Project Summary:The purpose of this study was to examine the metabolic profiles of mayfly (Centroptilum triangulifer) larvae subjected to a thermal challenge. This species is unusual in terms of its ease of culture, and its suitability as a laboratory test organism. Studies complementary to this metabolic study have focused on thermal effects on metabolic rates (respiration) and gene expression patterns in this species and do not support Prtners oxygen limitation paradigm. Our purpose here was to examine how an environmentally realistic thermal challenge affects the physiology of this organism.
Institute:North Carolina State University
Department:Environmental and Molecular Toxicology
Laboratory:Buchwalter Laboratory
Last Name:Buchwalter
First Name:David
Address:North Carolina State University Raleigh, NC 27695
Email:dbbuchwa@ncsu.edu
Phone:919-513-1129

Subject:

Subject ID:SU000042
Subject Type:Insect
Subject Species:Centroptilum triangulifer/Athorix diptoa/Megaloptera
Species Group:Insect

Factors:

Subject type: Insect; Subject species: Centroptilum triangulifer/Athorix diptoa/Megaloptera (Factor headings shown in green)

mb_sample_id local_sample_id species Treatment wing pads
SA001567131221ajcsa31_1Athorix Diptoa Athorix Diptoa control no wing pads
SA001568131221ajcsa17_1Athorix Diptoa Athorix Diptoa control no wing pads
SA001569131221ajcsa09_1Mayfly, Megalopterans, Athorix Diptoa Pooled QC sample no wing pads
SA001570131221ajcsa15_1Mayfly, Megalopterans, Athorix Diptoa Pooled QC sample no wing pads
SA001571131221ajcsa29_1Mayfly, Megalopterans, Athorix Diptoa Pooled QC sample no wing pads
SA001572131221ajcsa35_1Mayfly, Megalopterans, Athorix Diptoa Pooled QC sample no wing pads
SA001573131221ajcsa21_1Mayfly, Megalopterans, Athorix Diptoa Pooled QC sample no wing pads
SA001574131221ajcsa23_1Mayfly, Megalopterans, Athorix Diptoa Pooled QC sample no wing pads
SA001581131221ajcsa24_1Mayfly Mayfly control from different study no wing pads
SA001582131221ajcsa11_1Mayfly Mayfly control from different study no wing pads
SA001590131221ajcsa14_1Mayfly Mayfly control no wing pads
SA001591131221ajcsa33_1Mayfly Mayfly control no wing pads
SA001592131221ajcsa20_1Mayfly Mayfly control no wing pads
SA001593131221ajcsa25_1Mayfly Mayfly control no wing pads
SA001594131221ajcsa12_1Mayfly Mayfly control no wing pads
SA001595131221ajcsa26_1Mayfly Mayfly control no wing pads
SA001583131221ajcsa32_1Mayfly Mayfly controlX wing pads
SA001584131221ajcsa04_1Mayfly Mayfly controlX wing pads
SA001585131221ajcsa03_1Mayfly Mayfly controlX wing pads
SA001586131221ajcsa05_1Mayfly Mayfly controlX wing pads
SA001587131221ajcsa06_1Mayfly Mayfly controlX wing pads
SA001588131221ajcsa07_1Mayfly Mayfly controlX wing pads
SA001589131221ajcsa19_1Mayfly Mayfly controlX wing pads
SA001575131221ajcsa10_1Mayfly Mayfly Heat Treated no wing pads
SA001576131221ajcsa13_1Mayfly Mayfly Heat Treated no wing pads
SA001577131221ajcsa27_1Mayfly Mayfly Heat Treated no wing pads
SA001578131221ajcsa16_1Mayfly Mayfly Heat Treated no wing pads
SA001579131221ajcsa34_1Mayfly Mayfly Heat Treated no wing pads
SA001580131221ajcsa30_1Mayfly Mayfly Heat Treated no wing pads
SA001596131221ajcsa18_1Megalopterans Megalopterans control no wing pads
SA001597131221ajcsa28_1Megalopterans Megalopterans control no wing pads
Showing results 1 to 31 of 31

Collection:

Collection ID:CO000025
Collection Summary:whole insects were collected
Collection Protocol Comments:Original culture material was obtained from the Stroud Water Research Center (Avondale, PA). Larvae were reared at 22C and fed a periphytic diet. When larvae reached a suitable size, they were separated from their food source for 12 hours to evacuate gut contents. Larvae were then divided into two treatment groups a control group maintained at the culturing temperature of 22C, and a thermal challenge group. The thermal challenge group was subjected to a temperature increase at a rate of 1C per hour. This rate of temperature change is commonly observed in temperate streams. When the treatment temperature reached 30C, larvae from both control and treatment groups were flash frozen in liquid nitrogen in groups of 12-13 larvae. Each grouping of 12-13 larvae comprised a composite replicate for the metabolic profiling analysis. A total of 3 replicates each for the control and thermal treatments were used in this study. Some larvae had developed dark wing pads during the experiment. This is a signal that the larvae were very close to emerging from the aquatic larval phase to a winged sub-adult phase. Enough of these larvae were present in the control treatment to be removed from the control cohort and considered as an independent sample.
Sample Type:whole insect
Volumeoramount Collected:50 mg
Storage Conditions:-80C

Treatment:

Treatment ID:TR000043
Treatment Summary:Metabolomics profile of mayfly larvae exposed to heat
Treatment Protocol Comments:Heat Treated | Control | ControlX
Treatment:Abiotic
Animal Acclimation Duration:exposed to heat (22C to 30C ramp); fasted for 8 hours | control (fasted for 8 hours) | control (fasted for 6 hours)

Sample Preparation:

Sampleprep ID:SP000038
Sampleprep Summary:Insect samples homogenized in 50% acetonitrile at a concentration of 50 mg/mL. Sample dried and aliquot taken for NMR. Sample returned and extract processed for GC/MS analysis. Samples were derivatized using 2 step derivatization method (methoximation with methoxy amine followed by TMS derivatization with MSTFA).
Sampleprep Protocol Filename:RTI_DB_INSECT_Metabolomics_Procedure_March12_2014.docx
Processing Method:Homogenization and Solvent Removal with SpeedVac
Processing Storage Conditions:4C
Extraction Method:50% Acetonitrile
Extract Concentration Dilution:dried on SpeedVac
Extract Storage:-80C
Sample Resuspension:resuspended in D2O + Chenomix ISTD solution
Sample Derivatization:Methoximation with methoxy amine at 40 mg/ml in pyridine followed by TMS derivatization with MSTFA
Sample Spiking:FAMES (C8-C30) added to MSTFA as retention time markers

Combined analysis:

Analysis ID AN000045
Analysis type MS
Chromatography type GC
Chromatography system Leco Pegasus IV GC
Column Restek Rtx-5Sil MS (30 x 0.25mm, 0.25um)
MS Type EI
MS instrument type GC x GC-TOF
MS instrument name Leco Pegasus IV TOF
Ion Mode POSITIVE
Units peak area

Chromatography:

Chromatography ID:CH000026
Chromatography Summary:GC/MS analysis is based on Oliver Fiehn methods
Instrument Name:Leco Pegasus IV GC
Column Name:Restek Rtx-5Sil MS (30 x 0.25mm, 0.25um)
Column Pressure:10.4 psi
Column Temperature:50C(0.5 min)-330C(@20C/min)-330C(5 min)
Flow Gradient:constant flow
Flow Rate:1.0 mL/min
Injection Temperature:250C
Retention Index:FAMEs markers (C8-C30)
Retention Time:FAMEs markers (C8-C30)
Analytical Time:20 minutes
Oven Temperature:50C(0.5 min)-330C(@20C/min)-330C(5 min)
Transferline Temperature:280C
Sample Syringe Size:10 L
Chromatography Type:GC

MS:

MS ID:MS000040
Analysis ID:AN000045
Instrument Name:Leco Pegasus IV TOF
Instrument Type:GC x GC-TOF
MS Type:EI
Ion Mode:POSITIVE
Capillary Temperature:280C (transfer line)
Helium Flow:1.0 ml/min
Ion Source Temperature:250C
Ionization Energy:70 eV
Source Temperature:250C
Dataformat:netcdf
Resolution Setting:1200
Scan Range Moverz:50-800
Scanning:20 spectra/sec
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