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MB Sample ID: SA044177
Local Sample ID: | ESP-1060 |
Subject ID: | SU000827 |
Subject Type: | Human |
Subject Species: | Homo sapiens |
Taxonomy ID: | 9606 |
Species Group: | Human |
Select appropriate tab below to view additional metadata details:
Subject:
Subject ID: | SU000827 |
Subject Type: | Human |
Subject Species: | Homo sapiens |
Taxonomy ID: | 9606 |
Species Group: | Human |
Factors:
Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
---|---|---|---|---|
ESP-1060 | SA044177 | FL009015 | Control | CaseStatus |
ESP-1060 | SA044177 | FL009015 | Female | SEX |
ESP-1060 | SA044177 | FL009015 | over50 | AGE_DICH |
Collection:
Collection ID: | CO000821 |
Collection Summary: | Subjects included 50 cases and 50 controls from the Chronic Fatigue Initiative (CFI) Cohort (28) recruited at 4 sites across the US who met the 1994 CDC Fukuda and/or Canadian consensus criteria for ME/CFS. |
Sample Type: | Blood |
Blood Serum Or Plasma: | Plasma |
Treatment:
Treatment ID: | TR000841 |
Treatment Summary: | Subjects included 50 cases and 50 controls from the Chronic Fatigue Initiative (CFI) Cohort (28) recruited at 4 sites across the US who met the 1994 CDC Fukuda and/or Canadian consensus criteria for ME/CFS. |
Sample Preparation:
Sampleprep ID: | SP000834 |
Sampleprep Summary: | 1.Thaw each 20 μL plasma aliquot at room temperature (see Aliquoting TEDDY samples SOP). Once thawed (~10min) place liquid plasma samples on ice. 2. Add 225 μL cold “MeOH with QC mix” (see SOP “QC mix for LC-MS lipid analysis”). Keep MeOH on ice during extraction 3. Vortex each sample for 10s, keeping the rest on ice during all the extraction. 4. Add 750 μL of cold MTBE with 22:1 CE, keep MTBE on ice during extraction 5. Vortex for 10s 6. Shake for 6min at 4°C in the orbital mixer. 7. Add 188 μL room temperature LC/MS grade water. 8. Vortex for 20 s 9. Centrifuge for 2 min @ 14,000 rcf (12300 rpm) 10. Remove supernatant, splitting into two aliquots of 350 μL, keeping one at –20°C for backup 11. Dry samples to complete dryness in the speed vacuum concentration system |
Sampleprep Protocol Filename: | SP_SOP_Extraction_of_plasma.pdf |
Combined analysis:
Analysis ID | AN001274 |
---|---|
Analysis type | MS |
Chromatography type | Reversed phase |
Chromatography system | Waters Acquity |
Column | Waters Acquity BEH C18 (150 x 2.1mm,1.7um) |
MS Type | ESI |
MS instrument type | Triple quadrupole |
MS instrument name | ABI Sciex API 4000 QTrap |
Ion Mode | NEGATIVE |
Units | Concentration (nM) |
Chromatography:
Chromatography ID: | CH000890 |
Instrument Name: | Waters Acquity |
Column Name: | Waters Acquity BEH C18 (150 x 2.1mm,1.7um) |
Column Temperature: | 60 °C |
Flow Gradient: | 0 min 25% (B), 1 min 40% (B), 1-2.5 min 42% (B), 2.5-4.5 50% (B), 4.5-10.5 65% (B), 10.5-12.5 75% (B), 12.5-14 85% (B), 14-14.5 95% (B), 14.5-15 25% (B), 15-16 25% (B) |
Flow Rate: | 0.25 uL |
Sample Injection: | 5 µL |
Solvent A: | 100% water; 0.1% acetic acid |
Solvent B: | 90% acetonitrile/ 10% isopropanol |
Analytical Time: | 16 min |
Washing Buffer: | Ethyl Acetate |
Weak Wash Solvent Name: | 20% methanol, 10% isopropanol |
Weak Wash Volume: | 600 µL |
Strong Wash Solvent Name: | 50:50 Acetonitrile:Methanol |
Strong Wash Volume: | 600 µL |
Sample Loop Size: | 17 µL |
Randomization Order: | Excel generated |
Chromatography Type: | Reversed phase |
MS:
MS ID: | MS001167 |
Analysis ID: | AN001274 |
Instrument Name: | ABI Sciex API 4000 QTrap |
Instrument Type: | Triple quadrupole |
MS Type: | ESI |
Ion Mode: | NEGATIVE |