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MB Sample ID: SA025643
| Local Sample ID: | 141217actsa06_1 |
| Subject ID: | SU000516 |
| Subject Type: | Animal |
| Subject Species: | Canis lupus familiaris |
| Taxonomy ID: | 9615 |
| Genotype Strain: | Hound-type |
| Age Or Age Range: | Adult |
| Weight Or Weight Range: | 35-60 lbs |
| Gender: | Male/Female |
| Animal Inclusion Criteria: | no evidence of gastrointestinal illness or other disease |
| Species Group: | Mammal |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU000516 |
| Subject Type: | Animal |
| Subject Species: | Canis lupus familiaris |
| Taxonomy ID: | 9615 |
| Genotype Strain: | Hound-type |
| Age Or Age Range: | Adult |
| Weight Or Weight Range: | 35-60 lbs |
| Gender: | Male/Female |
| Animal Inclusion Criteria: | no evidence of gastrointestinal illness or other disease |
| Species Group: | Mammal |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| 141217actsa06_1 | SA025643 | FL006018 | rectum | Organ |
| 141217actsa06_1 | SA025643 | FL006018 | Female | Sex |
Collection:
| Collection ID: | CO000510 |
| Collection Summary: | Animals were euthanized for an unrelated study. Within 1-3 hours after euthanasia, a ventral midline approach was used to access the gastrointestinal tract. A longitudinal incision was made through the serosal surface at each site sampled to expose a length of the lumen. Disposable plastic spatulae were used to collect intestinal contents from each site. Duodenal samples were collected aborally to the cranial flexure. The ileum was identified by the antimesenteric artery. The colon was identified by location in situ and samples were collected from the transverse or descending colon. Rectal contents were collected from as caudally as possible while still sampling through the abdominal incision. |
| Sample Type: | Other (digesta/intestinal contents) |
| Collection Frequency: | Once |
| Collection Duration: | Once |
| Volumeoramount Collected: | Five aliquots of 50-100mg |
| Storage Conditions: | -80°C |
| Collection Vials: | 2 mL screw cap polypropylene vials |
| Storage Vials: | 2 mL screw cap polypropylene vials |
| Collection Tube Temp: | room temp |
Treatment:
| Treatment ID: | TR000530 |
| Treatment Summary: | The variable tested across the samples was the site of the GI tract, so none of the animals received any treatment for the purpose of comparing the effect of it to a control. The individual dogs are indicated by the letter A, C, D, E, F, and G. For each dog, we collected samples from duodenum, ileum, colon, and rectum. |
Sample Preparation:
| Sampleprep ID: | SP000523 |
| Sampleprep Summary: | 1. Weigh 50 mg tissue sample in to a 25 ml conical polypropylene centrifuge tube. 2. Add 2.5mL extraction solvent to the tissue sample and homogenize for 45 seconds ensuring that sample resembles a powder. In between samples, clean the homogenizer in solutions of methanol, acetone, water, and the extraction solvent. 3. 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. 4. Evaporate one 500μl aliquot of the sample in the Labconco Centrivap cold trap concentrator to complete dryness 5. The dried aliquot is then re-suspended with 500l 50% acetonitrile (degassed as given) 6. Centrifuge for 2 min at 14000 rcf using the centrifuge Eppendorf 5415. 7. Remove supernatant to a new Eppendorff tube. 8. Evaporate the supernatant to dryness in the the Labconco Centrivap cold trap concentrator. 9. Submit to derivatization. |
Combined analysis:
| Analysis ID | AN000761 |
|---|---|
| Analysis type | MS |
| Chromatography type | GC |
| Chromatography system | Leco Pegasus III GC |
| 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: | CH000547 |
| Methods Filename: | Data_Dictionary_Fiehn_laboratory_GCTOF_MS_primary_metabolism_10-15-2013_general.pdf |
| Instrument Name: | Leco Pegasus III GC |
| 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: | MS000673 |
| Analysis ID: | AN000761 |
| 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 |
