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MB Sample ID: SA078233
| Local Sample ID: | OPC-3uM-3 |
| Subject ID: | SU001196 |
| Subject Type: | Cultured cells |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
| Gender: | Not applicable |
| Cell Strain Details: | HepG2 cells |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU001196 |
| Subject Type: | Cultured cells |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
| Gender: | Not applicable |
| Cell Strain Details: | HepG2 cells |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| OPC-3uM-3 | SA078233 | FL011918 | 3 | Treatment |
Collection:
| Collection ID: | CO001190 |
| Collection Summary: | Culture medium of HepG2 cells in 100-mm dish was aspirated and cells were washed twice by 5% mannitol solution, and then processed with sampleprep steps. |
| Sample Type: | HepG2 cells |
| Storage Conditions: | Room temperature |
Treatment:
| Treatment ID: | TR001211 |
| Treatment Summary: | HepG2 cells were seeded into a 100mm dish the day before OPC-163493 treatment. OPC-163493 treatments (DMSO control, 1, 3, or 10uM, each n=2) were performed for 30min in FBS-free DMEM with high glucose (25mM). |
| Treatment Dose: | 0uM, 1uM, 3uM, 10uM |
| Treatment Doseduration: | 30 min |
| Cell Media: | MEM with Earle`s salts, L-Glutamine and Non-Essiontial Amino Acids, 10% fetal bovine serum, and 1mM sodium pyruvate solution |
Sample Preparation:
| Sampleprep ID: | SP001204 |
| Sampleprep Summary: | Culture medium of HepG2 cells in 100-mm dish was aspirated and cells were washed twice by 5% mannitol solution, and then the cells were treated with methanol. The cell extract was treated with Mili-Q water containing internal standard and filtered with 5-kDa cutoff filter. The filtrate was centrifugally concentrated and re-suspended in 50 µL of Milli-Q water. |
| Processing Storage Conditions: | Room temperature |
| Extract Storage: | -80℃ |
| Sample Resuspension: | 50 uL Mili-Q |
Combined analysis:
| Analysis ID | AN001860 | AN001861 |
|---|---|---|
| Analysis type | MS | MS |
| Chromatography type | CE | CE |
| Chromatography system | Agilent 7100 CE | Agilent 7100 CE |
| Column | Fused silica capillary, i.d. 50 μm × 80 cm | Fused silica capillary, i.d. 50 μm × 80 cm |
| MS Type | ESI | ESI |
| MS instrument type | Other | Triple quadrupole |
| MS instrument name | Agilent 6210 TOF | Agilent 6460 QQQ |
| Ion Mode | UNSPECIFIED | UNSPECIFIED |
| Units | Concentration (pmol/1000000 cells) | Concentration (pmol/1000000 cells) |
Chromatography:
| Chromatography ID: | CH001347 |
| Chromatography Summary: | capillary electrophoresis was connected with time-of-flight mass spectrometry (CE-TOFMS) for cation analysis and tandem mass spectrometry (CE-MS/MS) for anion. |
| Instrument Name: | Agilent 7100 CE |
| Column Name: | Fused silica capillary, i.d. 50 μm × 80 cm |
| Chromatography Type: | CE |
MS:
| MS ID: | MS001720 |
| Analysis ID: | AN001860 |
| Instrument Name: | Agilent 6210 TOF |
| Instrument Type: | Other |
| MS Type: | ESI |
| MS Comments: | The spectrometer was scanned from m/z 50 to 1,000. Peaks were extracted using automatic integration software MasterHands (Keio University, Tsuruoka, Japan) in order to obtain peak information including m/z, migration time for CE-TOFMS measurement (MT) and peak area. Signal peaks corresponding to isotopomers, adduct ions, and other product ions of known metabolites were excluded, and remaining peaks were annotated with putative metabolites from the HMT metabolite database based on their MTs and m/z values determined by TOFMS. The tolerance range for the peak annotation was configured at ±0.5 min for MT and ±10 ppm for m/z. In addition, peak areas were normalized against those of the internal standards and then the resultant relative area values were further normalized by sample amount. |
| Ion Mode: | UNSPECIFIED |
| MS ID: | MS001721 |
| Analysis ID: | AN001861 |
| Instrument Name: | Agilent 6460 QQQ |
| Instrument Type: | Triple quadrupole |
| MS Type: | ESI |
| MS Comments: | The spectrometer was scanned from m/z 50 to 1,000. Peaks were extracted using MasterHands, automatic integration software (Keio University, Tsuruoka, Yamagata, Japan) and MassHunter Quantitative Analysis B.04.00 (Agilent Technologies) in order to obtain peak information including m/z, peak area, and migration time (MT). Signal peaks were annotated according to the HMT metabolite database based on their m/z values with the MTs. The peak area of each metabolite was normalized with respect to the area of the internal standard and metabolite concentration was evaluated by standard curves with three-point calibrations using each standard compound. |
| Ion Mode: | UNSPECIFIED |
