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MB Sample ID: SA006634
| Local Sample ID: | H07 |
| Subject ID: | SU000140 |
| Subject Type: | Animal |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
| Genotype Strain: | C57BL/6J |
| Age Or Age Range: | 20 weeks |
| Weight Or Weight Range: | 26.8 +/- 2.2 g |
| Gender: | Male |
| Animal Animal Supplier: | Jackson Labs (Bar Harbor, ME) |
| Animal Housing: | 8 / cage |
| Animal Light Cycle: | 12 h / 12 h light/dark |
| Animal Feed: | Standard chow, ad lib. |
| Animal Water: | Ad lib. Access |
| Species Group: | Mammal |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU000140 |
| Subject Type: | Animal |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
| Genotype Strain: | C57BL/6J |
| Age Or Age Range: | 20 weeks |
| Weight Or Weight Range: | 26.8 +/- 2.2 g |
| Gender: | Male |
| Animal Animal Supplier: | Jackson Labs (Bar Harbor, ME) |
| Animal Housing: | 8 / cage |
| Animal Light Cycle: | 12 h / 12 h light/dark |
| Animal Feed: | Standard chow, ad lib. |
| Animal Water: | Ad lib. Access |
| Species Group: | Mammal |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| H07 | SA006634 | FL001648 | Heart | Tissue |
| H07 | SA006634 | FL001648 | Isoflurane overdose | Anesthesia / Euthanasia method |
Collection:
| Collection ID: | CO000124 |
| Collection Summary: | 4 tissues plus blood serum were collected and immediately following euthanasia or anesthesia induction, frozen in liquid nitrogen and stored at -80C until analysis |
| Sample Type: | Skeletal muscle (gastrocnemius), Liver, Heart, Epididymal white adipose (fat), blood serum |
| Collection Method: | Surgical dissection |
| Collection Time: | As soon as possible following anesthesia/euthanasia (all tissues collected within 3 minutes) |
| Storage Conditions: | -80 C |
Treatment:
| Treatment ID: | TR000142 |
| Treatment Summary: | Mice were prepared or tissue collection by one of 6 modes of anesthesia or euthanasia, then tissues were rapidly surgically collected and frozen |
| Animal Anesthesia: | One of 6 methods: 1) Cervical dislocation, 2) Carbon dioxide inhalation, 3) Isoflurane overdose, 4) continuous isoflurane, 5) ketamine, 6) pentobarbital |
| Animal Fasting: | 5 hours |
| Animal Endp Euthanasia: | All tissue collection were terminal studies; in methods 1-3, animals were euthanized prior to tissue collection; in methods 4-6, animals were under deep anesthesia while tissues were collected. |
Sample Preparation:
| Sampleprep ID: | SP000137 |
| Sampleprep Summary: | Frozen tissue samples were pulverized in a liquid nitrogen-chilled mortar and pestle, then sonicated in cold extraction solvent (7:2:1 methanol:chloroform:water) at an approximate ratio of 30 mg tissue (wet mass) /1 mL of solvent. After allowing samples to rest on ice 5 minutes, they were centrifuged to pellet precipitated protein and cell debris; the supernatant was placed in autosampler vials for direct LC-MS analysis. For serum, extraction solvent (1:1:1 methanol:acetonitrile:acetone) was added to serum at a 4:1 solvent:serum ratio, vortexed to mix, then centrifuged and supernatant recovered for LC-MS analysis. |
| Sampleprep Protocol Filename: | PROTOCOLS_mouse_anesthesia.pdf |
| Processing Method: | Cryo-pulverization + sonication in extraction solvent |
Combined analysis:
| Analysis ID | AN000203 |
|---|---|
| Analysis type | MS |
| Chromatography type | HILIC |
| Chromatography system | Agilent 1200 |
| Column | Phenomenex Luna NH2 (150 x 2.1mm,3um) |
| MS Type | ESI |
| MS instrument type | TOF |
| MS instrument name | Agilent 6220 TOF |
| Ion Mode | NEGATIVE |
| Units | Peak area |
Chromatography:
| Chromatography ID: | CH000135 |
| Chromatography Summary: | Polar metabolites were analyzed by HILIC-MS using an aminopropyl stationary phase |
| Instrument Name: | Agilent 1200 |
| Column Name: | Phenomenex Luna NH2 (150 x 2.1mm,3um) |
| Column Pressure: | 150 bar |
| Column Temperature: | 25 C |
| Flow Gradient: | 80-0% B over 20 minutes, hold 4 minutes, return to 80%B |
| Flow Rate: | 0.25 mL/min |
| Sample Injection: | 25 uL |
| Solvent A: | 100% water; 5 mM ammonium acetate, pH 9.9 |
| Solvent B: | 100% acetonitrile |
| Analytical Time: | 32 min |
| Chromatography Type: | HILIC |
MS:
| MS ID: | MS000166 |
| Analysis ID: | AN000203 |
| Instrument Name: | Agilent 6220 TOF |
| Instrument Type: | TOF |
| MS Type: | ESI |
| Ion Mode: | NEGATIVE |
| Capillary Voltage: | 3500 |
| Collision Gas: | N2 |
| Dry Gas Flow: | 10 L/min |
| Dry Gas Temp: | 350 C |
| Gas Pressure: | 20 psig |
