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MB Sample ID: SA137397
| Local Sample ID: | U_224 |
| Subject ID: | SU001695 |
| Subject Type: | Human |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
| Age Or Age Range: | 40-75 |
| Gender: | Male and female |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU001695 |
| Subject Type: | Human |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
| Age Or Age Range: | 40-75 |
| Gender: | Male and female |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| U_224 | SA137397 | FL017536 | User | Phenotype |
Collection:
| Collection ID: | CO001688 |
| Collection Summary: | Non-fasted urine sample; spot urine. |
| Sample Type: | Urine |
| Storage Conditions: | -80℃ |
Treatment:
| Treatment ID: | TR001708 |
| Treatment Summary: | N/A |
Sample Preparation:
| Sampleprep ID: | SP001701 |
| Sampleprep Summary: | Urine samples were allowed to thaw at 4 °C overnight. All samples were vortexed via multiple tube vortex for 2 min at 3000 rpm. The urine were spin downed using centrifuge at 1000 rpm for 30 sec. A volume of 7 µL aliquot of all 298 samples were transferred into one 10.0 mL tube labeled as pooled. A volume of 50 µL of urine (including study pool) was transferred to a new, pre-labeled Lo-Bind Eppendorf tube. A volume of 400 µL Protein Precipitation Buffer with Internal Standard (500 ng/ml Tryptophan-d5 in methanol) was added to each tube. All individual samples were thoroughly mixed on multiple tube vortexer for 2 mins at 5000 rpm. The protein precipitate was palleted using a centrifuge operating at room temperature and at 16,000 rcf for 4 min. A volume of 350 µl of the supernatant was transferred into pre-labeled 2.0 ml Low-bind Eppendorf tube. Sample were dried using speed vac overnight. A volume of 100 µL of reconstitution buffer (95:5 Water-Methanol) was added to each sample. Samples were thoroughly mixed on multiple tube vortexer for 10 mins at 5000 rpm. Samples were centrifuged at 4°C and at 16,000 rcf for 10 min. The supernatant was transferred to pre-labeled autosampler vials. A volume of 5 µL was injected onto column. |
| Processing Storage Conditions: | On ice |
| Extract Storage: | Described in summary |
Combined analysis:
| Analysis ID | AN002653 |
|---|---|
| Analysis type | MS |
| Chromatography type | Reversed phase |
| Chromatography system | Thermo Scientific™ Vanquish™ UPHPLC |
| Column | Waters ACQUITY UPLC HSS T3 (100 x 2.1mm,3um) |
| MS Type | ESI |
| MS instrument type | Orbitrap |
| MS instrument name | Thermo Q Exactive HF hybrid Orbitrap |
| Ion Mode | POSITIVE |
| Units | Peak area |
Chromatography:
| Chromatography ID: | CH001957 |
| Instrument Name: | Thermo Scientific™ Vanquish™ UPHPLC |
| Column Name: | Waters ACQUITY UPLC HSS T3 (100 x 2.1mm,3um) |
| Column Pressure: | 6000-10000 |
| Column Temperature: | 50 |
| Flow Gradient: | Time(min) Flow Rate %A %B Curve 1. 0 0.4 99.0 1.0 5 2. 1.00 0.4 99.0 1.0 5 3. 16.00 0.4 1.0 99.0 5 4. 19.00 0.4 1.0 99.0 5 5. 19.50 0.4 99.0 1.0 5 |
| Flow Rate: | 0.4 ml/min |
| Injection Temperature: | 8 |
| Solvent A: | 100% water; 0.1% formic acid |
| Solvent B: | 100% methanol; 0.1% formic acid |
| Chromatography Type: | Reversed phase |
MS:
| MS ID: | MS002461 |
| Analysis ID: | AN002653 |
| Instrument Name: | Thermo Q Exactive HF hybrid Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | Instrument: Thermo Q Exactive HFx Software: Progenesis QI 2.0 |
| Ion Mode: | POSITIVE |
| Capillary Temperature: | 275 °C |
| Capillary Voltage: | 3.5 KV |
| Collision Energy: | 10-35, ramp |
| Collision Gas: | N2 |
| Dry Gas Flow: | 45 |
| Dry Gas Temp: | 325°C |
| Fragmentation Method: | CID |
| Desolvation Gas Flow: | 45 |
