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MB Sample ID: SA258258
| Local Sample ID: | 5965 |
| Subject ID: | SU002670 |
| Subject Type: | Mammal |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
| Gender: | Male |
| Species Group: | Mammals |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU002670 |
| Subject Type: | Mammal |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
| Gender: | Male |
| Species Group: | Mammals |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| 5965 | SA258258 | FL033144 | High Fat diet | Diet |
| 5965 | SA258258 | FL033144 | Fasted | Nutritional status |
| 5965 | SA258258 | FL033144 | VLCAD +/+ | Genotype |
Collection:
| Collection ID: | CO002663 |
| Collection Summary: | Mice were sacrificed under anesthesia using ketamine (100mg/ml) and xylazine (20mg/ml). Immediately after mouse sacrifice, hearts were rapidly excised, snap-frozen with liquid nitrogen, and then stored at -80°C. |
| Sample Type: | Heart |
| Storage Conditions: | -80℃ |
| Collection Tube Temp: | -80 |
Treatment:
| Treatment ID: | TR002682 |
| Treatment Summary: | VLCAD-/- males (129svJ X C57BL/6J genetic background) were crossed with C57Bl/6J females to generate heterozygous VLCAD+/- mice. Coupling of heterozygous mice then produced VLCAD-/- mice and control littermate counterparts (VLCAD+/+). All animals were genotyped as previously described. Mice were housed in a specific pathogen-free facility under a 12-h light/dark cycle at constant temperature and with access to water and food ad libitum. At 7-month of age, mice were fed for 2 weeks with either a chow diet (Harlan no. 2018; 3.1 kcal/g) or a high-fat diet (Harlan Teklab no. 3584; 5.4kcal/g) ad libitum. Mice were sacrificed in fed state for the CD group (CD/fed condition) or after fasting for 24 hours for the HFD group (HFD/fasted condition. Percent calories from carbohydrates, lipids and proteins were 58/18/24 for the CD and 26.6/58.4/15 for the HFD. |
Sample Preparation:
| Sampleprep ID: | SP002676 |
| Sampleprep Summary: | Total lipids were extracted from 40mg of pulverized hearts under liquid nitrogen using in methyl tert-butyl ether (MTBE) and ethyl acetate after spiking with the following internal standards lysophosphatidylcholine (LPC) 13:0, phosphatidylcholine (PC) 19:0/19:0, PC14:0/14:0, phosphatidylserine (PS) 12:0/12:0, phosphatidylglycerol (PG) 15:0/15:0 and phosphatidylethanolamine (PE) 17:0/17:0). Supernatants were evaporated using a Speed Vacuum concentrator overnight, dissolved in methanol/chloroform (2:1) and aliquots (50 µL) were stored at -80ºC until use. |
| Sampleprep Protocol Filename: | Sample preparation protocol of untargeted lipidomic analysis |
| Processing Storage Conditions: | -80℃ |
Combined analysis:
| Analysis ID | AN004233 |
|---|---|
| Analysis type | MS |
| Chromatography type | Reversed phase |
| Chromatography system | Agilent 1290 Infinity |
| Column | Agilent ZORBAX Eclipse Plus C18 (100 x 2.1mm,1.8um) |
| MS Type | ESI |
| MS instrument type | QTOF |
| MS instrument name | Agilent 6530 QTOF |
| Ion Mode | POSITIVE |
| Units | Counts |
Chromatography:
| Chromatography ID: | CH003141 |
| Methods Filename: | LC-MS and data processing protocol of untargeted lipidomic analysis |
| Instrument Name: | Agilent 1290 Infinity |
| Column Name: | Agilent ZORBAX Eclipse Plus C18 (100 x 2.1mm,1.8um) |
| Column Temperature: | 40 |
| Flow Gradient: | 0–2 min, 50% B; 2–11 min, 50–85% B; 11–33 min, 85–94% B; 33–35 min, 94–98% B, 35–38 min; 98–99.5% B; 38–82 min, 98–99.5% B; 82–83 min, 99.5–50% B, plus an equilibration of 8 min |
| Flow Rate: | 0.45 ml/min |
| Solvent A: | 100% water; 0.2% formic acid; 10 mM ammonium formate |
| Solvent B: | 55% ethanol,/35% acetonitrile/10% MTBE, 55:35:10; 0.2% formic acid; 10 mM ammonium formate |
| Chromatography Type: | Reversed phase |
MS:
| MS ID: | MS003980 |
| Analysis ID: | AN004233 |
| Instrument Name: | Agilent 6530 QTOF |
| Instrument Type: | QTOF |
| MS Type: | ESI |
| MS Comments: | MS data processing was achieved using the Mass Hunter Qualitative Analysis software package (version B.07) for peak picking and a bioinformatic script that we have developed and encoded in both Perl and R languages to enable optimal MS data alignment, filtering of presence, normalisation, batch effect correction, and missing data imputation. The resulting final dataset list reproducible high quality MS signals referred as features with their mass-over-charge (m/z), corrected signal intensity, and retention time (RT). Feature identity (ID) was as following: ionization mode:mass@RT. |
| Ion Mode: | POSITIVE |
