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MB Sample ID: SA118752
| Local Sample ID: | 40001353_1 |
| Subject ID: | SU001496 |
| Subject Type: | Human |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
| Age Or Age Range: | 29-82 |
| Gender: | Male and female |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU001496 |
| Subject Type: | Human |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
| Age Or Age Range: | 29-82 |
| Gender: | Male and female |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| 40001353_1 | SA118752 | FL014844 | M | Sex |
| 40001353_1 | SA118752 | FL014844 | 325mg | Treatment |
Collection:
| Collection ID: | CO001491 |
| Collection Summary: | Blood plasma samples were collected from non-fasting participants in the Aspirin/Folate Polyp Prevention Study at enrollment (baseline) and after three years of study treatment. Blood samples were collected in tubes containing EDTA and after centrifugation aliquots of plasma were transferred to 1.8ml freezer tubes and stored frozen at -20C or lower. The samples were shipped on dry ice from the clinical centers to the Dartmouth biorepository storage facility (for storage at -70C) and subsequently to the metabolimics analysis lab at Emory University. |
| Sample Type: | Blood (plasma) |
| Storage Conditions: | Described in summary |
Treatment:
| Treatment ID: | TR001511 |
| Treatment Summary: | Samples were received frozen in aliquouts of <250uL. Prior to analysis, samples were thawed and prepared for HRM analysis using the standard protocols described in the Sample Preparation section. |
Sample Preparation:
| Sampleprep ID: | SP001504 |
| Sampleprep Summary: | Samples were prepared for metabolomics analysis using established methods(Johnson et al. (2010). Analyst; Go et al. (2015). Tox Sci). Prior to analysis, plasma aliquots were removed from storage at -80 degrees C and thawed on ice. Each cryotube was then vortexed briefly to ensure homogeneity, and 50 microliters was transferred to a clean microfuge tube. Immediately after, the plasma was treated with 100 microliters of ice-cold LC-MS grade acetonitrile (Sigma Aldrich) containing 2.5 microliters of internal standard solution with eight stable isotopic chemicals selected to cover a range of chemical properties. Following addition of acetonitrile, urine was equilibrated for 30min on ice, upon which precipitated proteins were removed by centrifuge (14,000rpm at 4 degrees C for 10 min). The resulting supernatant (100 microliters) was removed, added to a low volume autosampler vial and maintained at 4 degrees C until analysis (<22 h). |
| Sampleprep Protocol ID: | HRM_SP_082016_01 |
| Sampleprep Protocol Filename: | EmoryUniversity_HRM_SP_082016_01.pdf |
| Sampleprep Protocol Comments: | Date effective: 30 July 2016 |
| Extraction Method: | 2:1 acetonitrile: sample followed by vortexing and centrifugation |
Combined analysis:
| Analysis ID | AN002378 | AN002379 |
|---|---|---|
| Analysis type | MS | MS |
| Chromatography type | HILIC | Reversed phase |
| Chromatography system | Dionex UltiMate 3000 | Dionex UltiMate 3000 |
| Column | Thermo Fisher Accucore HILIC (50 x 2.1mm,2.6um) with Thermo accucore HILIC guard cartridge | Thermo Fisher Accucore C18 (50 x 2.1mm,2.6um) |
| MS Type | ESI | ESI |
| MS instrument type | Orbitrap | Orbitrap |
| MS instrument name | Thermo Q Exactive HF hybrid Orbitrap | Thermo Q Exactive HF hybrid Orbitrap |
| Ion Mode | POSITIVE | POSITIVE |
| Units | peak area | peak area |
Chromatography:
| Chromatography ID: | CH001745 |
| Chromatography Summary: | The HILIC column is operated parallel to reverse phase column for simultaneous analytical separation and column flushing through the use of a dual head HPLC pump equipped with 10-port and 6-port switching valves. During operation of HILIC separation method, the MS is operated in positive ion mode and 10 microliters of sample is injected onto the HILIC column while the reverse phase column is flushing with wash solution. Flow rate is maintained at 0.35 mL/min until 2 min, increased to 0.4 mL/min at 6 min and held for 4 min. Solvent A is 100% LC-MS grade water, solvent B is 100% LC-MS grade acetonitrile and solvent C is 2% formic acid (v/v) in LC-MS grade water. Initial mobile phase conditions are 25% A, 70% B, 5% C hold for 2 min, with linear gradient to 75% A, 20% B, 5% C at 6 min, hold for 4 min, resulting in a total analytical run time of 10 min. During the flushing phase (reverse phase analytical separation), the HILIC column is equilibrated with a wash solution of 25% A, 70% B, 5% C. |
| Methods ID: | 2% formic acid in LC-MS grade water |
| Methods Filename: | 20160120_posHILIC120kres10min_ESI_c18poswash_acrore.meth |
| Chromatography Comments: | Triplicate injections for each chromatography mode |
| Instrument Name: | Dionex UltiMate 3000 |
| Column Name: | Thermo Fisher Accucore HILIC (50 x 2.1mm,2.6um) with Thermo accucore HILIC guard cartridge |
| Column Temperature: | 60C |
| Sample Injection: | 10 uL |
| Solvent A: | 100% water |
| Solvent B: | 100% acetonitrile |
| Analytical Time: | 10 min |
| Sample Loop Size: | 15 uL |
| Sample Syringe Size: | 100 uL |
| Chromatography Type: | HILIC |
| Chromatography ID: | CH001746 |
| Chromatography Summary: | The C18 column is operated parallel to the HILIC column for simultaneous analytical separation and column flushing through the use of a dual head HPLC pump equipped with 10-port and 6- port switching valves. During operation of the C18 method, the MS is operated in negative ion mode and 10 uL of sample is injected onto the C18 column while the HILIC column is flushing with wash solution. Flow rate is maintained at 0.35 mL/min until 2 min, increased to 0.4 mL/min at 6 min and held for 4 min. Solvent A is 100% LC-MS grade water, solvent B is 100% LC-MS grade acetonitrile and solvent C is 2% formic acid (v/v) in LC-MS grade water. Initial mobile phase conditions are 80% A, 15% B, 5% C hold for 0.5 min, with linear gradient to 0% A, 95% B, 5% C at 2 min, hold for 4min, resulting in a total analytical run time of 10 min. During the flushing phase (HILIC analytical separation), the C18 column is equilibrated with a wash solution of 0% A, 95% B, 5% C until 7.5 min, followed by an equilibration solution of 80% A, 15% B, 5% C for 2.5 min. |
| Methods Filename: | 20160120_posC18120kres10mim_ESI_hilicposwash_acore.meth |
| Instrument Name: | Dionex UltiMate 3000 |
| Column Name: | Thermo Fisher Accucore C18 (50 x 2.1mm,2.6um) |
| Column Temperature: | 60C |
| Flow Rate: | 0.35 mL/min for 2 min; linear increase to 0.4 mL/min at 6 min held for 4 min |
| Sample Injection: | 10 uL |
| Solvent A: | 100% water |
| Solvent B: | 100% acetonitrile |
| Analytical Time: | 10 min |
| Sample Loop Size: | 15 uL |
| Sample Syringe Size: | 100 uL |
| Chromatography Type: | Reversed phase |
MS:
| MS ID: | MS002220 |
| Analysis ID: | AN002378 |
| Instrument Name: | Thermo Q Exactive HF hybrid Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | None |
| Ion Mode: | POSITIVE |
| Capillary Temperature: | 250C |
| Collision Gas: | N2 |
| Dry Gas Flow: | 45 |
| Dry Gas Temp: | 150C |
| Mass Accuracy: | < 3ppm |
| Spray Voltage: | 3500 |
| Activation Parameter: | 5.00E+05 |
| Activation Time: | 118ms |
| Interface Voltage: | S-Lens RF level= 55 |
| Analysis Protocol File: | EmoryUniversity_HRM_QEHF-MS_092017_v1.pdf |
| MS ID: | MS002221 |
| Analysis ID: | AN002379 |
| Instrument Name: | Thermo Q Exactive HF hybrid Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | None |
| Ion Mode: | POSITIVE |
| Capillary Temperature: | 250C |
| Collision Gas: | N2 |
| Dry Gas Flow: | 45 |
| Dry Gas Temp: | 150C |
| Mass Accuracy: | < 3ppm |
| Spray Voltage: | -4000 |
| Activation Parameter: | 5.00E+05 |
| Activation Time: | 118ms |
| Interface Voltage: | S-Lens RF level= 55 |
| Analysis Protocol File: | EmoryUniversity_HRM_QEHF-MS_092017_v1.pdf |
