Summary of Study ST000598

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 PR000436. The data can be accessed directly via it's Project DOI: 10.21228/M8PW36 This work is supported by NIH grant, U2C- DK119886.

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Study IDST000598
Study TitleDysfunctional lipid metabolism underlies the effect of the perinatal DDT exposure on the development of metabolic syndrome (part II)
Study SummaryThis study aims to identify changes in lipid mediators in the hypothalamus with triphenyl phosphate (TPP) exposure. UC Davis type 2 diabetes mellitus (UCD-T2DM) rats were treated with TPP (n=8 per group) or not treated (n=8 per group). Each group was analyzed for oxylipin, nitro lipids, endocannabinoid, and endocannabinoid-like monoacylglycerol and N-acylethanolamide changes to investigate alterations in lipid mediator signaling due to TPP exposure. Targeted metabolomic analysis of lipid mediators in rat hypothalamus samples was performed by the Newman lab.
Institute
University of California, Davis
DepartmentUSDA Western Human Nutrition Research Center
LaboratoryNewman Lab
Last NameNewman
First NameJohn
Address430 W. Health Sciences Dr., Davis, CA 95616
Emailjohn.newman@ars.usda.gov
Phone+1-530-752-1009
Submit Date2017-04-19
Raw Data AvailableYes
Raw Data File Type(s)wiff
Analysis Type DetailLC-MS
Release Date2017-07-10
Release Version1
John Newman John Newman
https://dx.doi.org/10.21228/M8PW36
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

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Sample Preparation:

Sampleprep ID:SP000628
Sampleprep Summary:Oxylipins and endocannabinoids were isolated using a Waters Ostro Sample Preparation Plate (Milford, MA). Hypothalamus samples were pulverized and aliquoted (~20-25mg) were added to 2mL polypropylene tubes and spiked with a 5 µL anti-oxidant solution (0.2 mg/ml solution BHT/EDTA in 1:1 MeOH:water) and 10 μL 1000nM analytical deuterated surrogates. A total of 100 µL methanol was added to the sample and vrtexed 90 sec. Next, 500 µL D.I. water and 1000 µL ethyl acetate was added and the tube was vortexed 3 minutes, before being centrifuged at 15,000g for 10 min at room temp. The supernate was then transferred into a clean 2 mL autosampler vial. The extraction with ethyl acetate was repeated and the eluent was dried by speed vacuum for 35 min at the medium BP setting. Once dry, samples were re-constituted with the internal standard 1-cyclohexyl ureido, 3-dodecanoic acid (CUDA) and 1-Phenyl 3-Hexadecanoic Acid Urea (PHAU) at 100 nM (50:50 MeOH:CAN), vortexed 1 min, transferred to a spin filter (0.1 µm, Millipore, Billerica, MA), centrifuged for 3 min at 6ºC at <4500g (rcf), before being transferred to 2 mL LC-MS amber vials. Extracts were stored at -20ºC until analysis by UPLC-MS/MS. The internal standard was used to quantify the recovery of surrogate standards.
Sampleprep Protocol Filename:Hypothal_Newman_Data_Report.docx
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