Return to study ST001148 main page
MB Sample ID: SA079795
| Local Sample ID: | 10268_23 |
| Subject ID: | SU001213 |
| Subject Type: | Cultured cells |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
| Age Or Age Range: | 21 |
| Gender: | Male |
| Cell Biosource Or Supplier: | Mayo Clinic |
| Cell Strain Details: | 10268 |
| Cell Passage Number: | 10 |
Select appropriate tab below to view additional metadata details:
Combined analysis:
| Analysis ID | AN001893 | AN001894 | AN001895 |
|---|---|---|---|
| Analysis type | MS | MS | MS |
| Chromatography type | GC | Reversed phase | Reversed phase |
| Chromatography system | Agilent 7890B | Waters Acquity | Waters Acquity |
| Column | Agilent HP5-MS (30m x 0.25mm, 0.25 um) | Waters Acquity BEH C18 (150 x 2mm,1.7um) | Waters Acquity BEH C8 (150 x 2.1mm,1.7um) |
| MS Type | EI | ESI | ESI |
| MS instrument type | Single quadrupole | Triple quadrupole | Triple quadrupole |
| MS instrument name | Agilent 5977A | Thermo Quantiva | Thermo Quantum Ultra |
| Ion Mode | POSITIVE | POSITIVE | POSITIVE |
| Units | micromol metabolite per gram protein | micromol metabolite per gram protein | micromol metabolite per gram protein |
Chromatography:
| Chromatography ID: | CH001370 |
| Chromatography Summary: | Organic acids were detected with an Agilent 5977A gas chromatography/mass spectrometry (GC/MS) under electron impact and single ion monitoring conditions. Analytes were separated on an Agilent DB-5MS column (30 m x 0.25 mm x 0.25μm). Sample injection volume was 1 μL performed in splitless mode. The inlet temperature was maintained at 250°C. The carrier gas was helium set at a flow rate of 0.9 ml/min. The initial oven temperature was 120°C set with the following ramp rates: Ramp to 180°C at 25°C/min; Ramp to 270°C at 6°C/min; Ramp to 325°C at 30°C/min. The transfer line temperature was 280°C. Concentrations of lactic acid, fumaric acid, succinic acid, oxaloacetic acid, alpha-ketoglutaric acid, malic acid, 2-hydroxyglutaric acid, cis-aconitic acid, citric acid, and isocitric acid were measured against a calibration curve. |
| Instrument Name: | Agilent 7890B |
| Column Name: | Agilent HP5-MS (30m x 0.25mm, 0.25 um) |
| Chromatography Type: | GC |
| Chromatography ID: | CH001371 |
| Chromatography Summary: | Amino acids were analyzed using Thermo TSQ Quantum Ultra mass spectrometer (West Palm Beach, FL) coupled with a Waters ACQUITY ultra performance liquid chromatography BEH C18 column (2.1 mm x 150 mm x. 1.7 μm). Data acquisition was performed using selected reaction monitoring (SRM) and positive electrospray ionization (ESI). Injection volume was 1 μL. The column flow rate was 400 μL/min with an isothermal set at 43°C. Mobile phase A was 1% acetonitrile in 0.1% formic acid, and mobile phase B was 100% acetonitrile. The mass spectrometer was operated with 4000 capillary voltage, 50 sheath gas, 20 auxiliary gas, and 15 sweep gas. The capillary temperature was 2700°C. Collision gas was 1.5 Torr and collision energy was 25 V. The tube lens was kept at 90 V. The concentration of amino acids was calculated against a calibration curve. |
| Instrument Name: | Waters Acquity |
| Column Name: | Waters Acquity BEH C18 (150 x 2mm,1.7um) |
| Column Temperature: | 43 |
| Flow Rate: | 400ul/min |
| Solvent A: | 99% water/1% acetonitrile; 0.1% formic acid |
| Solvent B: | 100% acetonitrile |
| Chromatography Type: | Reversed phase |
| Chromatography ID: | CH001372 |
| Chromatography Summary: | Acylcarnitines were analyzed using a Waters ACQUITY UPLC system (Milford, MA) coupled with a Thermo TSQ Quantiva tandem mass spectrometer (West Palm Beach, FL) in SRM and positive (H)ESI mode. Analytes were separated on a Waters ACQUITY UPLC BEH C8 column (2.1 mm x 150 mm x. 1.7 μm) with an isothermal temperature of 43°C. The mass spectrometer capillary voltage was set to 4000 with a sheath gas 30, auxiliary gas 5, and sweep gas 2. The ion transfer tube was maintained at 300°C with the vaporizer at 40°C, collision gas at 1.5 Torr, and collision energy at 12 V. Concentrations of carnitine, acetylcarnitine, propionylcarnitine, butyrylcarnitine, isovalerylcarnitine, octanoylcarnitine, lauroylcarnitine, myristoylcarnitine, palmitoylcarnitine, oleoylcarnitine, and stearoylcarnitine were measured against a calibration curve. |
| Instrument Name: | Waters Acquity |
| Column Name: | Waters Acquity BEH C8 (150 x 2.1mm,1.7um) |
| Chromatography Type: | Reversed phase |
