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MB Sample ID: SA255549
| Local Sample ID: | NTH5_023 |
| Subject ID: | SU002640 |
| Subject Type: | Cultured cells |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU002640 |
| Subject Type: | Cultured cells |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| NTH5_023 | SA255549 | FL032710 | HIGH GLUCOSE WITHOUT DRUGS | Treatment |
Collection:
| Collection ID: | CO002633 |
| Collection Summary: | hPSC-Schwann cells were treated with 5mM and 30mM glucose for 72 h and harvested for metabolomics analysis. |
| Collection Protocol Filename: | majd-et-al-csc-protocol.pdf |
| Sample Type: | iPSC cells |
Treatment:
| Treatment ID: | TR002652 |
| Treatment Summary: | Cells were treated with/without 30 mM glucose with/without bupropion hydrochloride. |
Sample Preparation:
| Sampleprep ID: | SP002646 |
| Sampleprep Summary: | Frozen total cell pellets from at least three biological repeats were submitted to the West Coast Metabolomics Center at the University of California, Davis. Agilent 7890C with Leco Pegasus HT was used for the study. Samples extracted using 1mL of 3:3:2 ACN:IPA:H2O (v/v/v). Half of the sample was dried to completeness and then derivatized using 10 uL of 40 mg/mL of Methoxyamine in pyridine. They were shaken at 30C for 1.5 hours. Then 91 uL of MSTFA + FAMEs to each sample and they were shaken at 37C for 0.5 hours to finish derivatization. Samples were then vialed, capped, and injected onto the instrument. A 7890C GC was used coupled with a LECO TOF. 0.5 uL of derivatized sample was injected using a splitless method onto a RESTEK RTX-5SIL MS column with an Intergra-Guard at 275C with a helium flow of 1 mL/min. The GC oven was set to hold at 50C for 1 min then ramped to 20C/min to 330C and then held for 5 min. The transfer line was set to 280C while the EI ion source was set to 250C. The Mass spec parameters collect data from 85m/z to 500m/z at an acquisition rate of 17 spectra/sec. |
| Sampleprep Protocol Filename: | majd-et-al-csc-protocol.pdf |
Combined analysis:
| Analysis ID | AN004185 |
|---|---|
| Analysis type | MS |
| Chromatography type | GC |
| Chromatography system | Agilent 7890C with Leco Pegasus HT |
| Column | Restek Rtx-5Sil (30m x 0.25mm,0.25um) |
| MS Type | EI |
| MS instrument type | GC x GC-TOF |
| MS instrument name | Agilent 7890C |
| Ion Mode | POSITIVE |
| Units | Peak height |
Chromatography:
| Chromatography ID: | CH003101 |
| Chromatography Summary: | Column name: Restek corporation Rtx-5Sil MS (30 m length x 0.25 mm internal diameter with 0.25 μm film made of 95% dimethyl/5%diphenylpolysiloxane) |
| Methods Filename: | majd-et-al-csc-protocol.pdf |
| Instrument Name: | Agilent 7890C with Leco Pegasus HT |
| Column Name: | Restek Rtx-5Sil (30m x 0.25mm,0.25um) |
| Column Temperature: | The GC oven was set to hold at 50C for 1 min then ramped to 20C/min to 330C and then held for 5 min |
| Flow Gradient: | helium flow |
| Flow Rate: | 1 mL/min |
| Solvent A: | 1mL of 3:3:2 ACN:IPA:H2O (v/v/v) |
| Solvent B: | 10 uL of 40 mg/mL of Methoxyamine in pyridine then Then 91 uL of MSTFA + FAMEs |
| Chromatography Type: | GC |
MS:
| MS ID: | MS003932 |
| Analysis ID: | AN004185 |
| Instrument Name: | Agilent 7890C |
| Instrument Type: | GC x GC-TOF |
| MS Type: | EI |
| MS Comments: | N/A. We performed the experiment at UC Davic Metabolomics Core facility. |
| Ion Mode: | POSITIVE |
| Analysis Protocol File: | majd-et-al-csc-protocol.pdf |
