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MB Sample ID: SA137117
Local Sample ID: | 9--3 |
Subject ID: | SU001691 |
Subject Type: | Mammal |
Subject Species: | Mus musculus |
Taxonomy ID: | 10090 |
Genotype Strain: | BALB/c |
Age Or Age Range: | 70-74 |
Gender: | Male and female |
Animal Animal Supplier: | Montana State University Animal Resource Center |
Animal Feed: | PicoLab Rodent Diet 20 5053 |
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Subject:
Subject ID: | SU001691 |
Subject Type: | Mammal |
Subject Species: | Mus musculus |
Taxonomy ID: | 10090 |
Genotype Strain: | BALB/c |
Age Or Age Range: | 70-74 |
Gender: | Male and female |
Animal Animal Supplier: | Montana State University Animal Resource Center |
Animal Feed: | PicoLab Rodent Diet 20 5053 |
Factors:
Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
---|---|---|---|---|
9--3 | SA137117 | FL017520 | High Dose Ricin | Treatment |
9--3 | SA137117 | FL017520 | 8h | Timepoint |
Collection:
Collection ID: | CO001684 |
Collection Summary: | Livers were removed from mice immediately following sacrificed, and stored at -80° C. Before metabolite extraction, livers were homogenized into powder with mortar and pestle over liquid nitrogen. |
Sample Type: | Liver |
Storage Conditions: | -80℃ |
Treatment:
Treatment ID: | TR001704 |
Treatment Summary: | Mice were injected via intraperitoneal (IP) route with either ricin or saline in 10 µL per g body weight. Mice in all but the fasted group were given food and water ad libitum. Fasted mice were transferred to new cages with fresh bedding where they had food withheld while given water ad libitum. Mice were sacrificed at 2hr (ricin), 8 hr (ricin and fasted), and overnight (16-22 hr, control, ricin, fasted). |
Treatment Route: | Intraperitoneal |
Treatment Dose: | 0.6 µg/kg body weight or 30 µg/kg body weight |
Treatment Vehicle: | saline |
Sample Preparation:
Sampleprep ID: | SP001697 |
Sampleprep Summary: | Frozen livers were homogenized in liquid nitrogen using a mortar and pestle. Liver powder was weighed (60-80 mg) into 2-mL microcentrifuge tubes. 900 µL of ice-cold 1:2 MeOH/CHCl3 was added to the liver tissue and tissue was mixed into suspension and lysed using a FastPrep-24TM 5G homogenizer using the S. aureus setting of 6.0 meters/second speed for 2 x 40 seconds. Tubes were placed back on ice and 200 µL of ice-cold H2O was added, before repeating the lysis step. Samples were then placed at -20 ºC for approximately 45 minutes to precipitate protein, then centrifuged at 14,000 g for 10 minutes. 250 µL of upper polar phase were transferred to a 1.5 mL Eppendorf tube and dried overnight using a Savant™ Universal SpeedVac™ vacuum system with no heat. Dried metabolite mixtures were re-suspended in 600 µL of NMR buffer containing 0.25 mM 4,4-dimethyl-4-silapentane-1-sulfonic acid (DSS), 90%H2O/10% D2O, 25 mM sodium phosphate, pH 7. |
Analysis:
MB Sample ID: | SA137117 |
Analysis ID: | AN002649 |
Laboratory Name: | Copie Lab |
Analysis Type: | NMR |
Num Factors: | 9 |
Num Metabolites: | 60 |
Units: | µM/mg liver tissue |
NMR:
NMR ID: | NM000186 |
Analysis ID: | AN002649 |
Instrument Name: | Bruker Avance III Solution NMR Spectrometer |
Instrument Type: | FT-NMR |
NMR Experiment Type: | 1D-1H |
Spectrometer Frequency: | 600.13 MHz (1H Larmor Frequency) |
NMR Probe: | 5 mm liquid-helium-cooled TCI cryoprobe with Z-gradient |
NMR Solvent: | 0.25 mM 4,4-dimethyl-4-silapentane-1-sulfonic acid (DSS), 90%H2O/10% D2O, 25 mM sodium phosphate, pH 7 |
NMR Tube Size: | 5 mm |
Shimming Method: | topshim |
Pulse Sequence: | zgesgp |
Number Of Scans: | 256 |
Num Data Points Acquired: | 64K |