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MB Sample ID: SA185669
| Local Sample ID: | OUHSC-22 |
| Subject ID: | SU002064 |
| Subject Type: | Cultured cells |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
| Gender: | Female |
| Cell Biosource Or Supplier: | OVCAR4 cell line was obtained from Dr. Thomas Hamilton (Fox Chase Cancer Center, PA) and OVCAR8 cells were from National Cancer Institute (NCI). Kuramochi, TYKNU, OVKATE and OVSAHO cell lines were from the JCRB Cell Bank, Tokyo, Japan. SNU119, SNU251, ES-2, OVCAR3, CAOV3 and OV90 cells were from Seoul National University, Seoul, Korea, and COV362, OAW28 and COV318 cells were purchased from Sigma-Aldrich (St. Louis, MO). |
| Cell Passage Number: | 3 and 4 |
| Cell Counts: | 20 million cells |
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Combined analysis:
| Analysis ID | AN003234 |
|---|---|
| Analysis type | MS |
| Chromatography type | Reversed phase |
| Chromatography system | Waters ACQUITY ultra-performance liquid chromatography (UPLC) |
| Column | Waters UPLC BEH C18 (100 x 2.1mm,1.7um) |
| MS Type | ESI |
| MS instrument type | Orbitrap |
| MS instrument name | Thermo Q Exactive HF hybrid Orbitrap |
| Ion Mode | UNSPECIFIED |
| Units | ratio |
MS:
| MS ID: | MS003008 |
| Analysis ID: | AN003234 |
| Instrument Name: | Thermo Q Exactive HF hybrid Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | All methods utilized a Waters ACQUITY ultra-performance liquid chromatography (UPLC) and a Thermo Scientific Q-Exactive high resolution/accurate mass spectrometer interfaced with a heated electrospray ionization (HESI-II) source and Orbitrap mass analyzer operated at 35,000 mass resolution. The sample extract was dried then reconstituted in solvents compatible to each of the four methods. Each reconstitution solvent contained a series of standards at fixed concentrations to ensure injection and chromatographic consistency. One aliquot was analyzed using acidic positive ion conditions, chromatographically optimized for more hydrophilic compounds. In this method, the extract was gradient eluted from a C18 column (Waters UPLC BEH C18-2.1x100 mm, 1.7 µm) using water and methanol, containing 0.05% perfluoropentanoic acid (PFPA) and 0.1% formic acid (FA). Another aliquot was also analyzed using acidic positive ion conditions, however it was chromatographically optimized for more hydrophobic compounds. In this method, the extract was gradient eluted from the same afore mentioned C18 column using methanol, acetonitrile, water, 0.05% PFPA and 0.01% FA and was operated at an overall higher organic content. Another aliquot was analyzed using basic negative ion optimized conditions using a separate dedicated C18 column. The basic extracts were gradient eluted from the column using methanol and water, however with 6.5mM Ammonium Bicarbonate at pH 8. The fourth aliquot was analyzed via negative ionization following elution from a HILIC column (Waters UPLC BEH Amide 2.1x150 mm, 1.7 µm) using a gradient consisting of water and acetonitrile with 10mM Ammonium Formate, pH 10.8. The MS analysis alternated between MS and data-dependent MSn scans using dynamic exclusion. The scan range varied slighted between methods but covered 70-1000 m/z. Raw data files are archived and extracted using proprietary, Metabolon LIMS data structures and analysis software (Metabolon Inc, Durham). Briefly, raw data was extracted, peak-identified and QC processed using Metabolon’s hardware and software. Biochemical identifications are based on three criteria: retention index within a narrow RI window of the proposed identification, accurate mass match to the library +/- 10 ppm, and the MS/MS forward and reverse scores between the experimental data and authentic standards. |
| Ion Mode: | UNSPECIFIED |
