Return to study ST000244 main page
MB Sample ID: SA011301
| Local Sample ID: | 2226 |
| Subject ID: | SU000264 |
| Subject Type: | Animal |
| Subject Species: | Equus caballus |
| Taxonomy ID: | 9796 |
| Genotype Strain: | Arabian |
| Species Group: | Mammal |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU000264 |
| Subject Type: | Animal |
| Subject Species: | Equus caballus |
| Taxonomy ID: | 9796 |
| Genotype Strain: | Arabian |
| Species Group: | Mammal |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| 2226 | SA011301 | FL004929 | EMS | Treatment |
| 2226 | SA011301 | FL004929 | Female | Gender |
Collection:
| Collection ID: | CO000255 |
| Collection Summary: | A mobile set-up allowed for immediate processing of samples from study horses on their home farm. 50mL of blood was collected into two EDTA tubes, two no-additive tubes, and one sodium oxalate tube. EDTA and sodium oxalate treated samples were thoroughly mixed by inversion and immediately centrifuged to pellet the cells and allow for removal of plasma by pipette. Blood drawn in a no-additive tube was allowed to clot, centrifuged, and had the serum removed by pipette. The draw yielded 6-8 mL of plasma and 3-4mL of serum, which was then divided in to 1mL aliquots and immediately snap frozen by immersion in liquid nitrogen. Upon returning to the lab, the samples were then transfered to -80C for storage. Currently, only plasma is being submitted for metabolomic analysis. |
| Collection Protocol Filename: | collection.txt |
| Sample Type: | Blood |
| Collection Method: | jugular venipuncture |
| Collection Location: | On farm |
| Collection Duration: | 30 minutes per horse total |
| Collection Time: | na |
| Volumeoramount Collected: | 50ml whole blood |
| Storage Conditions: | snap frozen in LN2 on farm, -80 in lab |
| Collection Vials: | glass vacutainer |
| Storage Vials: | plastic cryovials |
| Collection Tube Temp: | ambient during draw and centrifugation, then LN2 after plasma removal |
| Additives: | EDTA |
| Blood Serum Or Plasma: | PLASMA |
Treatment:
| Treatment ID: | TR000275 |
| Treatment Summary: | Blood was collected from the jugular vein and processed for serum, plasma, and DNA using standard procedures. To minimize circadian and meal associated effects on blood parameters, samples were collected in the morning prior to feeding concentrated rations. Care was taken to minimize stress to the animals, and no special treatments were given. Body measures were also gathered on each animal to capture regional adiposity and body size using a well optimized protocol, as well as body condition scores according to the Henneke system [numerical scores of 1 for emaciated to 9 for extremely obese]. Detailed diets, exercise regimes and a complete health history on each individual were recorded. Study animals are privately-owned, live on several farms in the central Florida area, and may have naturaly aquired disease. Based on the collected observations each horse is assigned to one of three diagnostic groups: 1) Normal Control, 2)Equine Metabolic Syndrome and 3)EMS- under treatment. |
| Treatment Protocol Filename: | treatment.txt |
Sample Preparation:
| Sampleprep ID: | SP000269 |
| Sampleprep Summary: | - |
| Sampleprep Protocol Filename: | Metabolomics_LCMSProtocol.pdf |
| Sampleprep Protocol Comments: | Standard protein precipitation method is included in this document |
| Processing Method: | PLASMA |
| Extraction Method: | PLASMA |
Combined analysis:
| Analysis ID | AN000381 | AN000382 |
|---|---|---|
| Analysis type | MS | MS |
| Chromatography type | Reversed phase | Reversed phase |
| Chromatography system | Thermo Dionex Ultimate 3000 | Thermo Dionex Ultimate 3000 |
| Column | ACE Excel 2 C18-PFP (100 x 2.1mm, 2um) | ACE Excel 2 C18-PFP (100 x 2.1mm, 2um) |
| MS Type | ESI | ESI |
| MS instrument type | Orbitrap | Orbitrap |
| MS instrument name | Thermo Q Exactive Orbitrap | Thermo Q Exactive Orbitrap |
| Ion Mode | POSITIVE | NEGATIVE |
| Units | peak area | peak area |
Chromatography:
| Chromatography ID: | CH000274 |
| Methods Filename: | Metabolomics_LCMSProtocol.pdf |
| Instrument Name: | Thermo Dionex Ultimate 3000 |
| Column Name: | ACE Excel 2 C18-PFP (100 x 2.1mm, 2um) |
| Flow Rate: | 0.350mL/min-0.600mL/min |
| Internal Standard: | Appendix A - Internal Standard Prep GLCMS.pdf |
| Solvent A: | 100% water; 0.1% formic acid |
| Solvent B: | 100% acetonitrile |
| Chromatography Type: | Reversed phase |
MS:
| MS ID: | MS000327 |
| Analysis ID: | AN000381 |
| Instrument Name: | Thermo Q Exactive Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| Ion Mode: | POSITIVE |
| MS ID: | MS000328 |
| Analysis ID: | AN000382 |
| Instrument Name: | Thermo Q Exactive Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| Ion Mode: | NEGATIVE |
