Summary of Study ST000357
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 PR000285. The data can be accessed directly via it's Project DOI: 10.21228/M82W27 This work is supported by NIH grant, U2C- DK119886.
See: https://www.metabolomicsworkbench.org/about/howtocite.php
| Study ID | ST000357 |
| Study Title | Broad Spectrum MS analysis of mouse hypothalmus from Anxiety Prone HSV-Latently Infected Obese Mice |
| Study Type | Broad Spectrum LCMS |
| Study Summary | Brain tissue samples for metabolomics experiments were generated in the following manner: 3-week old mice were placed on a 10% low fat (LF) diet and acclimated for one week prior to intranasal HSV-1 infection or mock infection with PBS. Fourteen days post-infection mice were randomized to either a 45% high fat diet (HF) or remained on the LF diet. Eight weeks post-diet transition, mice were euthanized and brain tissue samples were collected and processed for metabolomics. |
| Institute | University of North Carolina |
| Department | Discovery Sciences |
| Laboratory | Sumner Lab |
| Last Name | Sumner |
| First Name | Susan |
| Address | Eastern Regional Comprehensive Metabolomics Resource Core, UNC Nutrition Research Institute, 500 Laureate Way, Kannapolis, NC, 28081 |
| susan_sumner @unc.edu | |
| Phone | 704-250-5066 |
| Submit Date | 2016-03-03 |
| Num Groups | 4 |
| Total Subjects | 31 |
| Num Males | 31 |
| Raw Data Available | Yes |
| Raw Data File Type(s) | raw(Waters) |
| Analysis Type Detail | LC-MS |
| Release Date | 2017-03-03 |
| Release Version | 1 |
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Collection:
| Collection ID: | CO000372 |
| Collection Summary: | Brain tissue samples were homogenized in 50:50 Acetonitrile:Water (10 µL to every 1 mg of tissue) with washed ceramic beads on a MagNA Lyser, using two 30 sec pulses at 2,000 rpm with a 1 min chilling step in between pulses. Samples were vortexed on a multi-tube vortexer for 2 min at 5,000 rpm and centrifuged at room temperature for 4 min at 16,000 rcf. A 50 µL aliquot of each experimental brain tissue homogenized supernatant was transferred to a labeled 2.0 mL Lo-Bind eppendorf tube. Analytical pooled samples were created by combining 15 µL aliquot from each experimental sample in a 2.0 mL Lo-Bind eppendorf tube. The QC pooled sample was vortexed for 30 sec and 50 µL aliquots were transferred to 5 labeled 2.0 mL Lo-Bind eppendorf tubes. Next, 350 µL of acetonitrile was added to study sample and QC pooled sample tubes. The samples were vortexed on a multi-tube vortexer for 2 min at 5,000 rpm, and centrifuged at room temperature for 4 min at 16,000 rcf. The supernatants were transferred to new labeled tubes and dried on a lyophilizer overnight. Dried samples were reconstituted in 50 µL of 95:5 Water:Methanol, mixed on an analog vortex mixer for 1 min with speed set at 10. Then, the samples were centrifuged at room temperature for 4 min at 16,000 rcf, and the supernatants were transferred to autosampler vials for data acquisition. UPLC-MS Methods: UPLC-MS spectra were collected for all samples. UPLC was performed on a Waters Acquity UPLC with an Acquity HSS T3 column (2.1x 100mm x 1.8 µm) at 50 °C using the reversed-phase separation. Water with 0.1% formic acid (mobile phase A) and methanol with 0.1% formic acid (mobile phase B) were used as mobile phases and the metabolites were chromatographically separated using a gradient separation: (see the 3. Sheridan-Mice-Hypothalamus_RP-Metadata and Analytical Metadata.xlsx file for the flow gradient). Mass spectroscopy analysis was performed using a Synapt G2 Si ESI-Q-TOF using a 10 µL injection volume. UPLC-MS data were collected over 70-1000 m/z in both positive and negative modes. |
| Sample Type: | brain homogenate |
