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MB Sample ID: SA126707
| Local Sample ID: | 90_Old_Liver_24a |
| Subject ID: | SU001581 |
| Subject Type: | Mammal |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
| Genotype Strain: | C57BL/6JOlaHsd |
| Age Or Age Range: | 6 months (young) and 20-24 months (old) |
| Gender: | Male |
| Animal Animal Supplier: | Envigo |
| Animal Housing: | Animal rooms were maintained at 21°C-24°C and 35%-75% relative humidity |
| Animal Light Cycle: | 12/12-h (7am to 7pm) dark-light cycle |
| Animal Feed: | Altromin 1324 (chow) |
| Animal Water: | Ad lib |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU001581 |
| Subject Type: | Mammal |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
| Genotype Strain: | C57BL/6JOlaHsd |
| Age Or Age Range: | 6 months (young) and 20-24 months (old) |
| Gender: | Male |
| Animal Animal Supplier: | Envigo |
| Animal Housing: | Animal rooms were maintained at 21°C-24°C and 35%-75% relative humidity |
| Animal Light Cycle: | 12/12-h (7am to 7pm) dark-light cycle |
| Animal Feed: | Altromin 1324 (chow) |
| Animal Water: | Ad lib |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| 90_Old_Liver_24a | SA126707 | FL015583 | Old | Age |
| 90_Old_Liver_24a | SA126707 | FL015583 | Wild-type | Genotype |
| 90_Old_Liver_24a | SA126707 | FL015583 | N/A | Treatment |
Collection:
| Collection ID: | CO001576 |
| Collection Summary: | Mice were euthanized by cervical dislocation, left liver lobe was collected as fast as possible (<30 sec) and immediately flash-frozen in liquid nitrogen. Tissues were stored at -80C until metabolite extraction. |
| Sample Type: | Liver |
| Storage Conditions: | -80℃ |
Treatment:
| Treatment ID: | TR001596 |
| Treatment Summary: | All mice were fasted for 6h (8AM-2PM). Before sacrifice, tail blood was collected from all mice for less than 2 minutes. The untreated group were sacrificed at this point. For 13C-lactate treatment, 6h fasted mice were intraperitoneally injected with 1mg/kg [U-13C]Lactate (CIL) and sacrificed 15 minutes after injection. |
| Treatment: | [U-13C]Lactate injection |
| Treatment Compound: | [U-13C]Lactate |
| Treatment Route: | Intraperitoneal injection |
| Treatment Dose: | 1mg/kg |
| Treatment Dosevolume: | 300ul for 30gr mouse |
| Animal Anesthesia: | N/A |
| Animal Fasting: | 6h morning fast |
| Animal Endp Euthanasia: | Cervical dislocation |
Sample Preparation:
| Sampleprep ID: | SP001589 |
| Sampleprep Summary: | To maximize accuracy and to take into account tissue inhomogeneity, metabolites were extracted from 3 separate liver pieces for each mouse, and the average metabolite value of the 3 technical replicates was calculated at analysis. In detail, frozen liver pieces weighting ~30mg were transferred into soft tissue homogenizing CK 14 tubes containing 1.4 mm ceramic beads (Bertin corp.) prefilled with 1 ml of cold (-20°C) metabolites extraction solvent (Methanol: Acetonitrile:H2O, 50:30:20) and kept on ice. Samples were homogenized using precellys 24 tissue homogenizer (3X20sec at 6000 rpm, with a 30sec gap between each of the three cycles, Bertin Technologies) cooled to 2°C. Homogenized extracts were centrifuged in the Precellys tubes at 18,000g for 15 minutes at 4°C, supernatants were collected in microcentrifuge tubes and centrifuged again at 18,000g for 10 minutes at 4°C. The supernatants were transferred to glass HPLC vials and kept at -75 °C prior to LC-MS analysis. |
Combined analysis:
| Analysis ID | AN002498 |
|---|---|
| Analysis type | MS |
| Chromatography type | HILIC |
| Chromatography system | Thermo Dionex Ultimate 3000 |
| Column | SeQuant ZIC-HILIC (150 x 2.1mm,5um) |
| MS Type | ESI |
| MS instrument type | Orbitrap |
| MS instrument name | Thermo Q Exactive Orbitrap |
| Ion Mode | UNSPECIFIED |
| Units | Peak Area |
Chromatography:
| Chromatography ID: | CH001827 |
| Instrument Name: | Thermo Dionex Ultimate 3000 |
| Column Name: | SeQuant ZIC-HILIC (150 x 2.1mm,5um) |
| Column Temperature: | 45 |
| Flow Rate: | 0.2 ml/min |
| Solvent A: | 100% water; 20 mM ammonium carbonate adjusted to pH 9.2; 0.1% of 25% ammonium hydroxide |
| Solvent B: | 100% acetonitrile |
| Chromatography Type: | HILIC |
MS:
| MS ID: | MS002318 |
| Analysis ID: | AN002498 |
| Instrument Name: | Thermo Q Exactive Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | polarity switching mode to enable both positive and negative ions was used |
| Ion Mode: | UNSPECIFIED |
