Summary of Study ST001807

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 PR001141. The data can be accessed directly via it's Project DOI: 10.21228/M8711S 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 IDST001807
Study TitleUntargeted metabolomics of Daphnia magna exposed to a lithium cobalt oxide nanomaterial
Study TypeUntargeted MS
Study SummaryThe goal of this project was to determine lithium cobalt oxide (LCO)’s effects on pathways in the model organism Daphnia magna through untargeted metabolomics.
Institute
University of Wisconsin - Milwaukee
DepartmentSchool of Freshwater Sciences
LaboratoryRebecca Klaper
Last NameKlaper
First NameRebecca
Address600 E Greenfield Ave, Milwaukee, WI 53204
Emailrklaper@uwm.edu
Phone4143821713
Submit Date2021-05-18
Num Groups4
Total Subjects36
Study CommentsPooled samples of 20 Daphnia magna per replicate.
Raw Data AvailableYes
Raw Data File Type(s)mzXML
Analysis Type DetailMS(Dir. Inf.)
Release Date2022-05-19
Release Version1
Rebecca Klaper Rebecca Klaper
https://dx.doi.org/10.21228/M8711S
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

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Sample Preparation:

Sampleprep ID:SP001890
Sampleprep Summary:all solvents were chilled to 4 ºC. A mixture of 320 µL of HPLC grade MeOH and 128 µL of HPLC grade H2O were added to each sample tube and kept on ice. Tubes were then placed in a Precellys 24 homogeniser for 2 × 10s bursts at 6400 rpm. The homogenised mixture was then transferred into a clean 1.8 mL glass vial (Fisher TUL 520 006 J) using a Pasteur pipette. 320 µL (32 µL/mg) CHCl3 (HPLC grade) and 160 µL (16 µL/mg) dH2O (HPLC grade) were then added to each vial. These vials were vortexed on full power for 15 s each to thoroughly mix polar and non-polar solvents. Vials were then left on ice for on ice for 10 min to allow initial phase separation. Vials were then centrifuged at 4000 rpm at 4 ºC for 10 min to ensure complete phase separation. Centrifuged vials were allowed to come to room temperature by setting them on the lab bench for 5 min. Samples were then visibly biphasic, with protein debris separating the upper (polar) and lower (non-polar) layers. A 500 uL Hamilton syringe was then used to remove the polar phase (~ 2 × 150 µL aliquots) into 2 clean 1.5 mL Eppendorf tubes (one for positive, one for negative ion analysis). Polar samples were then dried using a Speed Vac concentrator and stored at -80 °C until analysis. Sample Preparation and Direct Infusion Mass Spectrometry Metabolomics For positive ion analysis, 30 µL of 4 ºC 80:20 methanol:water plus 0.25% formic acid was added to each of the frozen, dried extracts, and each sample vortexed for 30s. For negative ion analysis 30 µL of 4 ºC 80:20 methanol:water plus 20 mM ammonium acetate was added to each of the frozen, dried extracts, and each sample vortexed for 30 s. Samples were then centrifuged at 4000 g at 4 ºC for 10 mins. For both positive and negative ion analyses, samples were randomized and 5 µL of sample supernatant was pipetted into a pre-washed 96-well sample plate in quadruplicate. Three quality control (QC) samples (a mixture with equal volume from all samples) and a blank were also included on each plate. Loaded plates were covered with a foil seal using heat sealer and loaded into a TriVersa Nano-Mate® nanoelectro-spray ion source (Advion) with the cooler set at 10 ºC. Non-targeted analysis was carried out on polar fractions by direct infusion mass spectrometry (DIMS) using an LTQ Orbitrap Elite (Thermo Fisher Scientific). 21 overlapping selected ion monitoring (SIM) windows were collected covering m/z values from 50 to 620.
Processing Storage Conditions:4℃
Extract Storage:4℃
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