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MB Sample ID: SA214442
| Local Sample ID: | Control_Negative_OP_40 |
| Subject ID: | SU002330 |
| Subject Type: | Human |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU002330 |
| Subject Type: | Human |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| Control_Negative_OP_40 | SA214442 | FL026029 | Control | Group |
Collection:
| Collection ID: | CO002323 |
| Collection Summary: | Venous blood was drawn in 8.5 mL purple cap Vacutainer EDTA tubes and gently invert to mix. Plasma was collected after centrifugation at 1000 g for 15 min. All samples were immediately aliquoted to Eppendorf tubes and frozen at -80 ºC until analysis. |
| Sample Type: | Blood (plasma) |
| Storage Conditions: | -80℃ |
Treatment:
| Treatment ID: | TR002342 |
| Treatment Summary: | NA |
Sample Preparation:
| Sampleprep ID: | SP002336 |
| Sampleprep Summary: | 20 μL plasma, was mixed with 30 μL internal standard working solution, and extracted with 200 μL methanol for 10 min, and precipitated protein at -20℃ for 1h. After centrifugation at 14000xg for 10 min at 4 ℃, the supernatant was transferred into a new tube, dried under nitrogen, and resuspended in 50 μL acetonitrile/water (75/25, v/v). 5 μL of each sample was combined to make a pooled quality control (QC) sample and ran every ten samples in the long sequence to monitor retention time and signal intensity drift. The remaining samples (30 μL) were transferred into injection vials for metabolomic analysis. |
Combined analysis:
| Analysis ID | AN003663 | AN003664 |
|---|---|---|
| Analysis type | MS | MS |
| Chromatography type | HILIC | HILIC |
| Chromatography system | Thermo Dionex Ultimate 3000 | Thermo Dionex Ultimate 3000 |
| Column | Ascentis Express HILIC HPLC (150 x 2.1mm,2.7um) | Ascentis Express HILIC HPLC (150 x 2.1mm,2.7um) |
| 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 | NEGATIVE |
| Units | intensity | intensity |
Chromatography:
| Chromatography ID: | CH002714 |
| Instrument Name: | Thermo Dionex Ultimate 3000 |
| Column Name: | Ascentis Express HILIC HPLC (150 x 2.1mm,2.7um) |
| Column Temperature: | 35 |
| Flow Gradient: | 10% B for 2 min, and ramped to 30% B in 8 min, and ramped to 100 % B in 5 min, and maintained at 100 % B for 3 min, and back to initial 10 % B in 1 min, and equilibrate at initial condition for 7 min. |
| Flow Rate: | 0.25 ml/min |
| Injection Temperature: | 4 |
| Solvent A: | 95% acetonitrile/5% water; 0.1% acetic acid; 10 mM ammonium acetate |
| Solvent B: | 95% water/5% acetonitrile; 0.1% acetic acid; 10 mM ammonium acetate |
| Analytical Time: | 26min |
| Chromatography Type: | HILIC |
MS:
| MS ID: | MS003414 |
| Analysis ID: | AN003663 |
| Instrument Name: | Thermo Q Exactive HF hybrid Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | Samples were analyzed using a Q Exactive HF (QE-HF) (Thermo Scientific, Waltham, MA) equipped with a heated electro-spray ionization (HESI) source operated in both positive and negative ion mode. To build up metabolite spectral library for targeted analysis, we ran the pooled QC samples in full scan/ddMS2 mode (untargeted metabolomics). The Full Scan settings were as follows: AGC target, 1e6; Maximum IT, 200 ms; scan range, 60 to 900 m/z. Top 20 MS/MS spectral (dd-MS2) @ 15000 were generated with AGC target = 2e5, Maximum IT=25 ms, and (N)CE/stepped nce = 20, 30, 40v. Metabolites detection and identification were performed using Compound Discovery 2.1 (Thermo Scientific, Waltham, MA) by searching against online database (mzCloud) and in-house database built on Sigma metabolomics library (LSMLS, Sigma-Aldrich) (mzValut). Well annotated metabolites (MS spectral match and retention time match) were used to generate an inclusion list (Supplementary Table 3) for targeted PRM analysis. Final data acquisition was performed in Full scan + PRM modes. The PRM settings were as follows: resolution = 15000, AGC target = 2e5, Maximum IT=25 ms, loop count = 20, isolation window = 2.0, (N)CE was optimized for each metabolite. |
| Ion Mode: | POSITIVE |
| Capillary Temperature: | 325 |
| Spray Voltage: | 3500 |
| MS ID: | MS003415 |
| Analysis ID: | AN003664 |
| Instrument Name: | Thermo Q Exactive HF hybrid Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | Samples were analyzed using a Q Exactive HF (QE-HF) (Thermo Scientific, Waltham, MA) equipped with a heated electro-spray ionization (HESI) source operated in both positive and negative ion mode. To build up metabolite spectral library for targeted analysis, we ran the pooled QC samples in full scan/ddMS2 mode (untargeted metabolomics). The Full Scan settings were as follows: AGC target, 1e6; Maximum IT, 200 ms; scan range, 60 to 900 m/z. Top 20 MS/MS spectral (dd-MS2) @ 15000 were generated with AGC target = 2e5, Maximum IT=25 ms, and (N)CE/stepped nce = 20, 30, 40v. Metabolites detection and identification were performed using Compound Discovery 2.1 (Thermo Scientific, Waltham, MA) by searching against online database (mzCloud) and in-house database built on Sigma metabolomics library (LSMLS, Sigma-Aldrich) (mzValut). Well annotated metabolites (MS spectral match and retention time match) were used to generate an inclusion list (Supplementary Table 3) for targeted PRM analysis. Final data acquisition was performed in Full scan + PRM modes. The PRM settings were as follows: resolution = 15000, AGC target = 2e5, Maximum IT=25 ms, loop count = 20, isolation window = 2.0, (N)CE was optimized for each metabolite. |
| Ion Mode: | NEGATIVE |
| Capillary Temperature: | 325 |
| Spray Voltage: | 3000 |
