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MB Sample ID: SA019228
Local Sample ID: | 140603actsa32_3 |
Subject ID: | SU000422 |
Subject Type: | Animal |
Subject Species: | Rattus norvegicus |
Taxonomy ID: | 10116 |
Species Group: | Mammal |
Select appropriate tab below to view additional metadata details:
Subject:
Subject ID: | SU000422 |
Subject Type: | Animal |
Subject Species: | Rattus norvegicus |
Taxonomy ID: | 10116 |
Species Group: | Mammal |
Factors:
Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
---|---|---|---|---|
140603actsa32_3 | SA019228 | FL004719 | Duodenum | Organ |
140603actsa32_3 | SA019228 | FL004719 | Metformin 50mg/kg (oral administration) | Treatment |
Collection:
Collection ID: | CO000416 |
Collection Summary: | After 1 hour of treatment, blood was collected from the portal vein. Blood from the portal vein was cannulated with a fine syringe and blood was aspirated. Blood was spun down and plasma was collected and stored in -80. Urine was collected from bladder after 1 hour. Tissues (liver, kidney, intestine) were collected at the end of 1 hour. Only duodenum and ileum were used for metabolomics analysis. This study was conducted under the auspices of approved IACUC procedures in MuriGenics, Inc (Vallejo, CA). |
Sample Type: | Tissue |
Collection Location: | MuriGenics, Inc (Vallejo, CA) |
Collection Time: | 1 hour after treatment |
Treatment:
Treatment ID: | TR000436 |
Treatment Summary: | Twenty (20) adult rats (male, weighing approximately 200g) were randomized by body weights and assigned to five treatment groups (saline, aminoguanidine (positive control), 3 different oral doses of metformin). Before treatment, rats were starved for 24 hours. On treatment day, saline or aminoguanidine or metformin will be given to the rat. |
Animal Fasting: | 24 hours |
Sample Preparation:
Sampleprep ID: | SP000429 |
Sampleprep Summary: | 1. Weigh 4 mg tissue sample ( muscle 20 mg) in to a 2.0 ml eppendorf tube. Add 1.0 mL extraction solvent to the tissue sample and using GenoGrinder homogenize samples for 45 seconds ensuring that sample resembles a powder. 2. Centrifuge the samples at 2500 rpm. for 5 minutes. Aliquot 2 X 500µl supernatant, one for analysis and one for a backup sample. Store backup aliquot in the -20°C freezer. 3. Evaporate one 500µl aliquot of the sample in the Labconco Centrivap cold trap concentrator to complete dryness 4. The dried aliquot is then re-suspended with 500l 50% acetonitrile (degassed as given) (only for liver and brain samples). 5. Centrifuge for 2 min at 14000 rcf using the centrifuge Eppendorf 5415. 6. Remove supernatant to a new Eppendorff tube. 7. Evaporate the supernatant to dryness in the the Labconco Centrivap cold trap concentrator. 8. Submit to derivatization. |
Sampleprep Protocol Filename: | SOP_Extraction_of_Mammalian_Tissue_Samples.pdf |
Combined analysis:
Analysis ID | AN000640 |
---|---|
Analysis type | MS |
Chromatography type | GC |
Chromatography system | Agilent 6890N |
Column | Restek Corporation Rtx-5Sil MS |
MS Type | EI |
MS instrument type | GC-TOF |
MS instrument name | Leco Pegasus III GC TOF |
Ion Mode | POSITIVE |
Units | counts |
Chromatography:
Chromatography ID: | CH000465 |
Methods Filename: | Data_Dictionary_Fiehn_laboratory_GCTOF_MS_primary_metabolism_10-15-2013_general.pdf |
Instrument Name: | Agilent 6890N |
Column Name: | Restek Corporation Rtx-5Sil MS |
Column Pressure: | 7.7 PSI |
Column Temperature: | 50-330C |
Flow Rate: | 1 ml/min |
Injection Temperature: | 50 C ramped to 250 C by 12 C/s |
Sample Injection: | 0.5 uL |
Oven Temperature: | 50°C for 1 min, then ramped at 20°C/min to 330°C, held constant for 5 min |
Transferline Temperature: | 230C |
Washing Buffer: | Ethyl Acetate |
Sample Loop Size: | 30 m length x 0.25 mm internal diameter |
Randomization Order: | Excel generated |
Chromatography Type: | GC |
MS:
MS ID: | MS000572 |
Analysis ID: | AN000640 |
Instrument Name: | Leco Pegasus III GC TOF |
Instrument Type: | GC-TOF |
MS Type: | EI |
Ion Mode: | POSITIVE |
Ion Source Temperature: | 250 C |
Ionization Energy: | 70 eV |
Mass Accuracy: | Nominal |
Source Temperature: | 250 C |
Scan Range Moverz: | 85-500 Da |
Scanning Cycle: | 17 Hz |
Scanning Range: | 85-500 Da |
Skimmer Voltage: | 1850 V |