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MB Sample ID: SA338922
| Local Sample ID: | H_WTHFD_10 |
| Subject ID: | SU003243 |
| Subject Type: | Mammal |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU003243 |
| Subject Type: | Mammal |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| H_WTHFD_10 | SA338922 | FL040128 | Heart | Sample source |
| H_WTHFD_10 | SA338922 | FL040128 | WT | Genotype |
| H_WTHFD_10 | SA338922 | FL040128 | M | Sex |
| H_WTHFD_10 | SA338922 | FL040128 | High Fat Diet | Diet |
Collection:
| Collection ID: | CO003236 |
| Collection Summary: | Murine cardiac tissue was excised, then snap frozen in liquid nitrogen. |
| Sample Type: | Cardiac tissue |
Treatment:
| Treatment ID: | TR003252 |
| Treatment Summary: | CHCHD10 WT and CHCHD10 S55L heterozygous mice were treated with either a control diet (70% Carbohydrate, 20% Protein, 10% Fat) or High Fat Diet (60% Fat, 20% Protein, 20% Carbohydrate) in utero until 75 days of age. |
Sample Preparation:
| Sampleprep ID: | SP003250 |
| Sampleprep Summary: | 15 mg of cardiac tissue was homogenized in 80% methanol (Sigma) using Tissue Tearer (BioSpec) on dry ice. Samples were incubated at -80ºC for 4 hours. Homogenates were then centrifuged at 14,000 rfc for 20 min at 4ºC. The supernatant was extracted and stored at -80ºC for mass spectroscopy with the Weill Cornell Medicine Meyer Cancer Center Proteomics & Metabolomics Core Facility. |
Combined analysis:
| Analysis ID | AN005125 |
|---|---|
| Analysis type | MS |
| Chromatography type | HILIC |
| Chromatography system | Thermo Vanquish |
| Column | Merck SeQuant ZIC-pHILIC (150 x 2.1mm,5um) |
| MS Type | ESI |
| MS instrument type | Orbitrap |
| MS instrument name | Thermo Q Exactive Orbitrap |
| Ion Mode | UNSPECIFIED |
| Units | Peak Intensity |
Chromatography:
| Chromatography ID: | CH003878 |
| Methods Filename: | Protocol_HeartMetabolomics.pdf |
| Instrument Name: | Thermo Vanquish |
| Column Name: | Merck SeQuant ZIC-pHILIC (150 x 2.1mm,5um) |
| Column Temperature: | 30 |
| Flow Gradient: | 85% to 30% A in 20 min followed by a wash with 30% A and re-equilibration at 85% A |
| Flow Rate: | 150 μL/min |
| Solvent A: | 100% acetonitrile |
| Solvent B: | 100% water; 0.1% NH4OH; 20 mM CH3COONH4 |
| Chromatography Type: | HILIC |
MS:
| MS ID: | MS004861 |
| Analysis ID: | AN005125 |
| Instrument Name: | Thermo Q Exactive Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | The MS data was processed using XCalibur 4.1 (Thermo Scientific) to obtain the metabolite signal intensity for relative quantitation. Targeted identification was available for 205 metabolites based on an in-house library established using known chemical standards. Identification required exact mass (within 5ppm) and standard retention times. For untargeted metabolomics, metabolites were identified by mass matching of the MS signal to metabolites in the HMDB database. If multiple metabolites in the database were matched to a certain MS signal, all matched metabolites were grouped into a single identification, and ordered based on the number of references included in the HMDB database (high to low). We used the first ranked metabolite in the downstream analyses. When multiple values with the same metabolite attribution occurred (different metabolites with same mass and retention time), we opted to use all values in the analyses to avoid biases on which intensities/attributions to consider. Peak intensities for metabolites were screened for missing values and relative metabolite abundance data was analyzed by using MetaboAnalyst software version 5.0 (Pang et al, 2021). Metabolite significance was determined with one-way ANOVA with post-hoc t-tests, with the cutoff being a raw p value < 0.05, and the pathway significance cutoff was FDR corrected p value < 0.05. |
| Ion Mode: | UNSPECIFIED |
| Analysis Protocol File: | Protocol_HeartMetabolomics.pdf |
