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MB Sample ID: SA256620
Local Sample ID: | sample_0027 |
Subject ID: | SU002655 |
Subject Type: | Human |
Subject Species: | Homo sapiens |
Taxonomy ID: | 9606 |
Gender: | Not applicable |
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Subject:
Subject ID: | SU002655 |
Subject Type: | Human |
Subject Species: | Homo sapiens |
Taxonomy ID: | 9606 |
Gender: | Not applicable |
Factors:
Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
---|---|---|---|---|
sample_0027 | SA256620 | FL035347 | none | Factor |
Collection:
Collection ID: | CO002648 |
Collection Summary: | Fasting serum samples were collected for this study. None of the volunteers have undergone bariatric surgery or had taken antibiotics two months prior to their inclusion in the study. None of the participants were diagnosed with diabetes mellitus (Type 1 or 2). Thirty-four of study participants were taking statins at the time of study. The serum samples were stored at −80 °C until analyzed. |
Sample Type: | Blood (serum) |
Storage Conditions: | -80℃ |
Treatment:
Treatment ID: | TR002667 |
Treatment Summary: | Samples were stored at -80 as it is, and no treatment was done. |
Sample Preparation:
Sampleprep ID: | SP002661 |
Sampleprep Summary: | 40 µL of serum sample was mixed with 80 µL of acetonitrile including internal standards (TCA-d4, GUDCA-d4, GCA-d4, CA-d4, UDCA-d4, GCDCA-d4, CDCA-d4, DCA-d4, GLCA-d4 and LCA-d4; PFOA-13C8, PFNA-13C5, PFUndA-13C7, PFHxS-13C3 and PFOS-13C8, succinic acid-d4, glutamine-d5, valine-d8, tryptophan-d5, 3-hydroxybutyric acid-d4 and heptadecanoic acid-d5) and vortexed, ultrasonicated and centrifuged. 90 µL of the supernatant was evaporated to dryness and resuspended 60 µL of MeOH/H2O (70/30). |
Processing Storage Conditions: | -80℃ |
Extract Storage: | -80℃ |
Combined analysis:
Analysis ID | AN004206 |
---|---|
Analysis type | MS |
Chromatography type | Reversed phase |
Chromatography system | Agilent 1290 Infinity II |
Column | Waters ACQUITY UPLC BEH C18 (100 x 2.1mm,1.7um) |
MS Type | ESI |
MS instrument type | QTOF |
MS instrument name | Agilent 6545 QTOF |
Ion Mode | NEGATIVE |
Units | ng/ml |
Chromatography:
Chromatography ID: | CH003117 |
Instrument Name: | Agilent 1290 Infinity II |
Column Name: | Waters ACQUITY UPLC BEH C18 (100 x 2.1mm,1.7um) |
Column Temperature: | 50 |
Flow Gradient: | from min 0-1.5, the percentage of phase B was increased from 5% to 30%, from min 1.5-4.5, the percentage of B was increased to 70%, from min 4.5-7.5 the percentage of B was increased to 100% and held for 5.5 mins. A post-time of 5 min was used to regain the initial conditions for the next analysis. |
Flow Rate: | 0.4 mL/min |
Solvent A: | 70% water/30% methanol; 2mM ammonium acetate |
Solvent B: | 100% methanol; 2mM ammonium acetate |
Chromatography Type: | Reversed phase |
MS:
MS ID: | MS003953 |
Analysis ID: | AN004206 |
Instrument Name: | Agilent 6545 QTOF |
Instrument Type: | QTOF |
MS Type: | ESI |
MS Comments: | The analyses were done using UHPLC-QTOFMS instrument from Agilent Technologies (Santa Clara, CA, USA). The analysis was done on an ACQUITY UPLC® BEH C18 column (2.1 mm × 100 mm, particle size 1.7 µm) by Waters (Milford, MA, USA). Mobile phase A consisted of H2O:MeOH (v/v 70:30) and mobile phase B of MeOH with both phases containing 2mM ammonium acetate as an ionization agent. The LC pump was programmed at a flow rate of 0.4 mL/min with the elution gradient as follows: from min 0-1.5, the percentage of phase B was increased from 5% to 30%, from min 1.5-4.5, the percentage of B was increased to 70%, from min 4.5-7.5 the percentage of B was increased to 100% and held for 5.5 mins. A post-time of 5 min was used to regain the initial conditions for the next analysis. The total analysis time per sample was 18 min. The dual ESI ionization source was settings were as follows: capillary voltage was 4.5 kV, nozzle voltage 1500 V, N2 pressure in the nebulized was 21 psi and the N2 flow rate and temperature as sheath gas was 11 L/min and 379 °C, respectively. The m/z range 100-1700 in negative ion mode was used. MassHunter B.06.01 software (Agilent Technologies, Santa Clara, CA, USA) was used for all data acquisition. MS data processing was performed using open source software MZmine 2.5232. Quality control was performed throughout the dataset by including blanks, pure standard samples, and control plasma samples. The average RSD for the PFAS in QC samples was 10.6%, showing the robustness of the analytical procedure. The unknown compounds were identified based on exact mass and MS/MS data. |
Ion Mode: | NEGATIVE |