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MB Sample ID: SA126674
Local Sample ID: | 93_Old_Liver_25a |
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 |
---|---|---|---|---|
93_Old_Liver_25a | SA126674 | FL015581 | Old | Age |
93_Old_Liver_25a | SA126674 | FL015581 | SIRT6-tg | Genotype |
93_Old_Liver_25a | SA126674 | FL015581 | 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 |