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MB Sample ID: SA034202
| Local Sample ID: | 201_CFS |
| Subject ID: | SU000640 |
| Subject Type: | Human |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
| Age Or Age Range: | 53 ± 3.5 (23-69 yr) |
| Gender: | Male |
| Human Ethnicity: | White/non-Hispanic |
| Species Group: | Human |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU000640 |
| Subject Type: | Human |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
| Age Or Age Range: | 53 ± 3.5 (23-69 yr) |
| Gender: | Male |
| Human Ethnicity: | White/non-Hispanic |
| Species Group: | Human |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| 201_CFS | SA034202 | FL007476 | CFS | Treatment |
Collection:
| Collection ID: | CO000634 |
| Collection Summary: | Blood samples from 18 male subjects with chronic fatigue syndrome and 20 age and sex matched controls were collected. Plasma was prepared according to the standard protocol. The metabolites were extracted from |
| Sample Type: | Blood |
| Collection Method: | Lithium-heparin vacutainer tubes |
| Storage Conditions: | -80 C |
| Collection Vials: | Lithium-heparin vacutainer tubes |
| Collection Tube Temp: | -80 C |
| Blood Serum Or Plasma: | Plasma |
Treatment:
| Treatment ID: | TR000654 |
| Treatment Summary: | No treatment. There are only two groups: control and disease group (Chronic fatigue syndrome) |
Sample Preparation:
| Sampleprep ID: | SP000647 |
| Sampleprep Summary: | Metabolites from plasma were extracted using pre-chilled methanol/acetonitrile (50:50) |
| Extraction Method: | Typically, 90 μl of plasma was thawed on ice and mixed with 5 μl of isotope standards and incubated for 10 min at 20°C to permit small molecules and vitamins in the internal standards to associate with plasma binding proteins. Macromolecules (protein, DNA, RNA, glycans, etc.) were precipitated by extraction with 4 volumes (400 μl) of cold (-20°C), acetonitrile:methanol (50:50) and incubated on crushed ice for 10 min, then removed by centrifugation at 16,000g x 10 min at 4°C. The supernatants containing the extracted metabolites and internal standards were transferred to labeled cryotubes and stored at -80°C for LCMS/MS analysis. |
| Extract Storage: | -80 |
| Sample Derivatization: | No |
| Sample Spiking: | 25-35 commercial stable isotope internal standards, and 57 stable isotope internal standards that were custom-synthesized in E. coli and S. cerevisiae by metabolic labeling with 13C-glucose and 13C bicarbonate |
Combined analysis:
| Analysis ID | AN000946 | AN000947 |
|---|---|---|
| Analysis type | MS | MS |
| Chromatography type | HILIC | HILIC |
| Chromatography system | Shimadzu 20AD | Shimadzu 20AD |
| Column | Shodex NH2P-40 2E (250× 2.0 mm) | Shodex NH2P-40 2E (250× 2.0 mm) |
| MS Type | ESI | ESI |
| MS instrument type | Triple quadrupole | Triple quadrupole |
| MS instrument name | ABI Sciex 5500 QTrap | ABI Sciex 5500 QTrap |
| Ion Mode | NEGATIVE | POSITIVE |
| Units | AUC | AUC |
Chromatography:
| Chromatography ID: | CH000674 |
| Chromatography Summary: | hydrophilic interaction chromatography (HILIC) on shodex column |
| Instrument Name: | Shimadzu 20AD |
| Column Name: | Shodex NH2P-40 2E (250× 2.0 mm) |
| Column Pressure: | 900~2800 psi |
| Column Temperature: | 25 C |
| Flow Gradient: | 0-3.5 min: 95%B, 3.6-8 min: 85% B, 8.1-13 min: 75% B, 13.5–35 min: 0% B, 36–46 min: 95% B, 46.1 min: end. |
| Flow Rate: | 0.2 ml/min |
| Injection Temperature: | room temperature |
| Solvent A: | 95% water/5% acetonitrile; 20 mM ammonium carbonate; 38 mM ammonium hydroxide, pH 9.8 |
| Solvent B: | 100% acetonitrile |
| Analytical Time: | 46.1 min |
| Oven Temperature: | 25 C |
| Sample Loop Size: | 10 ul |
| Sample Syringe Size: | 100 ul |
| Chromatography Type: | HILIC |
MS:
| MS ID: | MS000842 |
| Analysis ID: | AN000946 |
| Instrument Name: | ABI Sciex 5500 QTrap |
| Instrument Type: | Triple quadrupole |
| MS Type: | ESI |
| Ion Mode: | NEGATIVE |
| Collision Energy: | Compound-specific |
| Collision Gas: | High |
| Ion Source Temperature: | 500 C |
| Ion Spray Voltage: | -4500 |
| Ionization: | ESI |
| Mass Accuracy: | 0.1 Da |
| Scanning: | MRM |
| MS ID: | MS000843 |
| Analysis ID: | AN000947 |
| Instrument Name: | ABI Sciex 5500 QTrap |
| Instrument Type: | Triple quadrupole |
| MS Type: | ESI |
| Ion Mode: | POSITIVE |
| Collision Energy: | Compound-specific |
| Collision Gas: | High |
| Ion Source Temperature: | 500 C |
| Ion Spray Voltage: | 5500 |
| Mass Accuracy: | 0.1 Da |
| Scanning: | MRM |
