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MB Sample ID: SA174788
Local Sample ID: | NIST SRM 1950-15V |
Subject ID: | SU001950 |
Subject Type: | Human |
Subject Species: | Homo sapiens |
Taxonomy ID: | 9606 |
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Subject:
Subject ID: | SU001950 |
Subject Type: | Human |
Subject Species: | Homo sapiens |
Taxonomy ID: | 9606 |
Factors:
Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
---|---|---|---|---|
NIST SRM 1950-15V | SA174788 | FL021465 | Blood | Source |
NIST SRM 1950-15V | SA174788 | FL021465 | 15V | MS2 CE |
Collection:
Collection ID: | CO001943 |
Collection Summary: | NIST SRM 1950 was purchased from NIST, and HepG2 cells were purchased from the Chinese Tissue Culture Collections (CTCC, Shanghai, China). Urine was collected from volunteers. Six- to eight-week-old male C57BL/6 mice were fed a regular standard breeding diet and housed under standard pathogen-free (SPF) conditions. Faecal samples and liver tissue were collected and immediately stored at -80 °C for further analysis. |
Sample Type: | Blood (plasma), Urine, HepG2 cells |
Storage Conditions: | -80℃ |
Treatment:
Treatment ID: | TR001962 |
Treatment Summary: | No treatment |
Sample Preparation:
Sampleprep ID: | SP001956 |
Sampleprep Summary: | Plasma (NIST SRM 1950): A 240 μL volume of Mix-IS-ACN was added to 60 μL of NIST SRM 1950 for protein precipitation. Then, the sample was vortexed for 60 s and centrifuged for 10 min at 15,000 g and 4 °C. Then, 250 μL of supernatant was transferred to a new centrifuge tube for lyophilization. The residue was reconstituted with 60 μL of 90% H2O/CH3OH (v/v). Urine: A 60 μL sample of urine was centrifuged for 10 min at 15,000 g and 4 °C, and the supernatant was transferred to a new centrifuge tube. Then, 30 μL Mix-IS-MeOH was added to the same centrifuge tube, and the mixture was thoroughly combined on a vortex mixer for 60 s. HepG2: HepG2 cells were purchased from the Chinese Tissue Culture Collections (CTCC, Shanghai, China) and cultured in Dulbecco’s modified Eagle’s medium (DMEM, Gibco) supplemented with 10% fetal bovine serum (FBS, Gibco), 100 IU/ml penicillin and 100 µg/ml streptomycin (Gibco). These cells were cultured at 37 °C in a humidified 5% CO2 atmosphere and treated within 24 hours after the confluence of cells reached 80%. The culture media of the cells was removed, and the cells were washed three times in phosphate-buffered saline (PBS). Then, the cells were quickly frozen in liquid nitrogen and stored at -80 °C before extraction. Then, 1 mL 80% CH3OH/H2O (v/v) was added to the cells, and the cells were scraped with extraction solvent. Both the extract and cell/debris suspension were transferred to a clean tube and thoroughly mixed on a vortex mixer for 120 s. Ultrasonication at 30 Hz for 2 × 10 s was performed. After being allowed to settle for 10 min, the cell extraction solution was centrifuged for 15 min at 15,000 g and 4 °C. Then, 800 μL of supernatant was transferred to a new centrifuge tube for lyophilization. The residue was reconstituted with 40 μL of Mix-IS-MeOH. |
Combined analysis:
Analysis ID | AN003035 |
---|---|
Analysis type | MS |
Chromatography type | Reversed phase |
Chromatography system | Waters Acquity |
Column | Waters Acquity BEH C8 (100 x 2.1mm,1.7um) |
MS Type | ESI |
MS instrument type | QTOF |
MS instrument name | ABI Sciex 5600+ TripleTOF |
Ion Mode | POSITIVE |
Units | peak area |
Chromatography:
Chromatography ID: | CH002248 |
Instrument Name: | Waters Acquity |
Column Name: | Waters Acquity BEH C8 (100 x 2.1mm,1.7um) |
Column Temperature: | 50 |
Flow Rate: | 0.35 mL/min |
Chromatography Type: | Reversed phase |
MS:
MS ID: | MS002822 |
Analysis ID: | AN003035 |
Instrument Name: | ABI Sciex 5600+ TripleTOF |
Instrument Type: | QTOF |
MS Type: | ESI |
MS Comments: | The spray voltage was 5.5 kV and the source temperature was 550 °C, Curtain gas, gas 1 and gas 2 were set at 35, 55, and 55 psi, respectively. The data was processed by XCMS, MS-DIAL, MZmine 2 and MetEx. |
Ion Mode: | POSITIVE |