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MB Sample ID: SA033562
Local Sample ID: | OHSUserum_PO6976_2 |
Subject ID: | SU000631 |
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: | SU000631 |
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 |
---|---|---|---|---|
OHSUserum_PO6976_2 | SA033562 | FL007454 | serum | Sample_Type |
OHSUserum_PO6976_2 | SA033562 | FL007454 | case | Factor |
Collection:
Collection ID: | CO000625 |
Collection Summary: | Matched DBS and serum samples were simultaneously collected in 2000–2001 from 10 overnight fasted male participants having an average age of 75.5 ± 6.15 years. The institutional review boards at the participating institutions approved the protocol and the study participants provided informed consent. Case participants included 5 diabetic men with high BMIs, TGs and glucose levels, and low amounts of HDL. The 5 control participants had normal metabolic profiles (normal BMIs, TGs, HDL and glucose levels). |
Sample Type: | Blood |
Storage Conditions: | The DBS were stored at room temperature in the dark until analysis, while the serum was stored at –80°C. |
Treatment:
Treatment ID: | TR000645 |
Treatment Summary: | Samples collected in 2000–2001 were stored until June 2015. |
Sample Preparation:
Sampleprep ID: | SP000638 |
Sampleprep Summary: | For the serum samples, 10 mL of whole blood was drawn and centrifuged so the serum could be extracted. For the DBS samples, 500 μL of blood was collected and 125 μL was blotted in four areas on a Whatman FTA Classic Card. Polar metabolites and lipid extracts were derived from the same sample type (DBS or serum) using a modified Folch extraction in June 2015 and analyzed with MS shortly after. The DBS punch from each patient was transferred into to a 2.0 mL tube where 50 μL of water and then 1200 μL of –20°C 2:1 chloroform/methanol were added. Each sample was vortexed for 30 s then transferred into a shaker at 22°C for 60 min at 600 rpm. The samples were vortexed again for 30 s and then 250 μL of water was added to induce a phase separation. The sample was gently inverted several times, placed at room temperature for 5 min and then centrifuged at 10,000 g for 5 min at 4°C and put on ice to maintain the clear phase separation. Finally, 400 μL of the top polar layer was removed, dried in a Speedvac, and stored at –80°C for analysis of polar metabolites, while 700 μL of the bottom nonpolar layer was removed, dried in a Speedvac, and stored at –20°C in 250 μL of 2:1 chloroform/methanol for lipid analyses. Serum lipids were extracted using a similar procedure except 25 μL of serum was used and then 600 μL of –20°C 2:1 chloroform/methanol was added. After vortexing and shaking, 125 μL of water was added to induce a phase separation and 200 μL of the top polar layer and 350 μL of the bottom nonpolar lipid layer were removed and stored as outlined above. Prior to analysis, the total lipid extracts were dried down and then reconstituted in 70 μL and 100 μL of MeOH for the DBS and serum samples, respectively. To generate pooled case and control samples for LC/MS/MS and LC/IMS-MS analyses, 5 μL aliquots from each reconstituted DBS and serum sample were removed and combined. |
Combined analysis:
Analysis ID | AN000929 | AN000930 | AN000931 |
---|---|---|---|
Analysis type | MS | MS | MS |
Chromatography type | Reversed phase | Reversed phase | GC |
Chromatography system | Waters NanoAcquity | Waters NanoAcquity | Agilent 7890A |
Column | Waters Acquity HSS T3 (150 x 1mm,1.8um) | Waters Acquity HSS T3 (150 x 1mm,1.8um) | Agilent HP5-MS (30m × 0.25mm, 0.25 um) |
MS Type | ESI | ESI | EI |
MS instrument type | Orbitrap | Orbitrap | Single quadrupole |
MS instrument name | Thermo Velos Pro Orbitrap | Thermo Velos Pro Orbitrap | Agilent 7890A |
Ion Mode | POSITIVE | NEGATIVE | POSITIVE |
Units | Peak Height | Peak | Peak Area |
Chromatography:
Chromatography ID: | CH000663 |
Instrument Name: | Waters NanoAcquity |
Column Name: | Waters Acquity HSS T3 (150 x 1mm,1.8um) |
Chromatography Type: | Reversed phase |
Chromatography ID: | CH000664 |
Instrument Name: | Agilent 7890A |
Column Name: | Agilent HP5-MS (30m × 0.25mm, 0.25 um) |
Chromatography Type: | GC |
MS:
MS ID: | MS000825 |
Analysis ID: | AN000929 |
Instrument Name: | Thermo Velos Pro Orbitrap |
Instrument Type: | Orbitrap |
MS Type: | ESI |
Ion Mode: | POSITIVE |
MS ID: | MS000826 |
Analysis ID: | AN000930 |
Instrument Name: | Thermo Velos Pro Orbitrap |
Instrument Type: | Orbitrap |
MS Type: | ESI |
Ion Mode: | NEGATIVE |
MS ID: | MS000827 |
Analysis ID: | AN000931 |
Instrument Name: | Agilent 7890A |
Instrument Type: | Single quadrupole |
MS Type: | EI |
Ion Mode: | POSITIVE |