Summary of Study ST000049

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.

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Study IDST000049
Study TitleMetabolomics Analysis of Thermally Challenged Mayfly Larvae (NMR 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 NMR Metabolomics could be used to distinguish among the different types of larvae.
Institute
University of North Carolina
DepartmentDiscovery Science Technology
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
Raw Data AvailableYes
Raw Data File Type(s)fid
Uploaded File Size7.6 M
Analysis Type DetailNMR
Release Date2015-05-01
Release Version1
Susan Sumner Susan Sumner
https://dx.doi.org/10.21228/M85P4V
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

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Sample Preparation:

Sampleprep ID:SP000063
Sampleprep Summary:Control and heat-treated mayfly samples were provided in triplicate and were processed individually. One sample was provided for AD, ControlX, MF control (from a previous study) and MG larvae. Three replicates of the larvae listed above having only a single sample were created. Aliquots of 50-300mg of insect larvae were mixed with degassed 1:1 Acetonitrile:Water solution in a 2 mL snap cap tube at a concentration of 50 mg/mL. Megalopterans were substantially larger than the other larvae and were prepared at 200 mg/mL. Samples were homogenized and then centrifuged at 4?C for 5 minutes at 14000rcf. The supernatant was removed and placed into a new tube. Samples were centrifuged again at 4?C for 5 minutes at 14000rcf and a volume of the homogenate corresponding to 40 mg insect larvae were taken. After homogenization, the three larval replicates from AD, ControlX, MG and MF control (from a previous study) were pooled and a single sample created for each larval type. There was insufficient Mayfly (heat and control) sample material to create an internal pool, so an external pool was made by combining equal concentration amounts of AD, MG and MF control (from a previous study) pool quality check (QC) samples. Samples were completely dried by vacuum centrifuge and were reconstituted by adding 630 µL of D2O (Aldrich) and 70 µL of Chenomx ISTD solution (Chenomx, Edmonton, Alberta, Canada). The samples were vortexed and centrifuged at 14000rcf for 2 minutes. 650 µL of sample was transferred into 5mm NMR tubes and analyzed by NMR.
Sampleprep Protocol Filename:RTI_Metabolomic_Analysis_of_Thermally_Challenged_Larvae_Procedure.docx
Processing Method:Homogenization and Solvent Removal with SpeedVac
Processing Storage Conditions:4°C
Extraction Method:50% Aqueous Acetonitrile
Extract Concentration Dilution:dried on SpeedVac
Extract Storage:-80C
Sample Resuspension:resuspended in D2O + Chenomx ISTD solution
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