Summary of study ST001124

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 PR000732. The data can be accessed directly via it's Project DOI: 10.21228/M82D79 This work is supported by NIH grant, U2C- DK119886.

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Study IDST001124
Study TitleEarly Mechanistic Events Induced by Secondhand Smoke Prevalent Low Molecular Weight Polycyclic Aromatic Hydrocarbons in Mouse Lung Epithelial Cells
Study SummaryWe evaluated lung epithelial cells exposed to low molecular weight polycyclic aromatic hydrocarbons and what lipid metabolites were produced following early exposure, prior to metabolism. For the targeted study (AKB study 2, we used 40uM dose of the 2PAHs (1methylanthracene and fluoranthene; 1:1 ratio)and assessed 2,4,8,and 12 hrs of treatment with the PAHs. We also examined a 24 h time point in another study at a lower dose (15 uM LMW PAH mixture; 1:1 ratio of 1-methanthrancene and fluoranthene).
Institute
University of Colorado, Denver
DepartmentAnschutz Medical Campus
Last NameBauer
First NameAlison
Address12850 E. Montview Dr. Rm 3125, Aurora, CO 80045
Emailalison.bauer@ucdenver.edu
Phone303-724-6297
Submit Date2018-11-06
Raw Data AvailableYes
Raw Data File Type(s).cd,.cg, .xml, .bin, .stg
Analysis Type DetailLC-MS
Release Date2019-02-27
Release Version1
Alison Bauer Alison Bauer
https://dx.doi.org/10.21228/M82D79
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

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Sample Preparation:

Sampleprep ID:SP001193
Sampleprep Summary:Briefly, before SPE cleanup, the frozen samples were removed from the freezer and centrifuged at 14,000 RPM at 4°C for 10 minutes. The supernatant was removed, transferred to a new microcentrifuge tube, and spiked with 10 μL of 10 pg/μL internal standard solution (100 pg of each: 5(S)-HETE-d8, 8-iso-PGF2a-d4, 9(S)-HODE-d4, LTB4-d4, LTD4-d5, LTE4-d5, PGE2-d4, PGF2a-d9 and RvD2-d5 in ethanol). The supernatant was dried in a vacuum centrifuge and reconstituted with 1.0 mL of 10% methanol. The reconstituted extracts were loaded on a Strata-X 33 μm 30 mg/1maL SPE column (Phenomenex, Torrance, California, USA) preconditioned with 1.0 mL of methanol followed by 1.0 mL of water. The SPE column was washed with 10% methanol and then eluted directly into a reduced surface activity/maximum recovery glass autosampler vial with 1.0 mL of methyl formate. The methyl formate was evaporated completely from the vial with a stream of nitrogen and the SPE cartridge was then eluted with 1.0 mL of methanol directly into the same autosampler vial. The methanol was evaporated to dryness with a stream of nitrogen and the sample was reconstituted with 20 uL of ethanol. The samples were analyzed immediately or frozen at -70 °C until analysis.
Sampleprep Protocol Filename:AKB_Targeted-protocol.docx
Sampleprep Protocol Comments:For AKB study 2 protocol, the collection, treatment, and sample prep are all in the same file.
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