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MB Sample ID: SA240119
| Local Sample ID: | #717_mito |
| Subject ID: | SU002492 |
| Subject Type: | Mammal |
| Subject Species: | Rattus norvegicus |
| Taxonomy ID: | 10116 |
Select appropriate tab below to view additional metadata details:
Combined analysis:
| Analysis ID | AN003917 |
|---|---|
| Analysis type | MS |
| Chromatography type | Reversed phase |
| Chromatography system | Agilent 1290 Infinity II |
| Column | Agilent ZORBAX Eclipse Plus C18 (100 x 2.1mm,1.8um) |
| MS Type | ESI |
| MS instrument type | Triple quadrupole |
| MS instrument name | Agilent 6490 QQQ |
| Ion Mode | POSITIVE |
| Units | pmol per mg |
MS:
| MS ID: | MS003656 |
| Analysis ID: | AN003917 |
| Instrument Name: | Agilent 6490 QQQ |
| Instrument Type: | Triple quadrupole |
| MS Type: | ESI |
| MS Comments: | Lipid extraction. Muscle homogenates/mitochondrial isolates were extracted using a modified single-phase chloroform/methanol extraction as described previously [Weir et al. 2016]. In brief, 20 volumes of chloroform:methanol (2:1) was added to the sample along with a series of internal standards. Samples were vortexed and spun on a rotoary mixer for 10 minutes. After sonication on a sonicator bath for 30 minutes, samples were rested for a further 20 minutes prior to centrifugation at 13,000 x g for 10 minutes. Supernatants were transferred into a 96 well plated, dried down and reconstituted in 50L water saturated butanol and sonicated for 10 minutes. After the addition of 50l of methanol with 10mM ammonium formate, the samples were spun down again at 4000RPM on a plate centrifuge (Heraeus multifuge 1S-R, ThermoFisher) and transferred into glass vials with inserts for mass spectrometry analysis. Targeted lipidomics analysis. Liquid chromatography tandem mass spectrometry (LC-MS/MS) was performed according to previously published methods, with slight modification for tissue samples [Huynh et al. 2019]. Sample extracts were analysed using either (i) a 4000 QTRAP mass spectrometer (Sciex) for cardiolipins as described preciously [Tan et al. 2020] or (ii) an Agilent 6490 QQQ mass spectrometer all other lipid species. Lipids run on the Agilent 6490 were measured using scheduled multiple reaction monitoring with the following conditions: Isolation widths for Q1 and Q3 were set to “unit” resolution (0.7 amu), gas temperature, 150°C, nebulizer 20psi, sheath gas temperature 200°C, gas flow rate 17L/min, capillary voltage 3500V and sheath gas flow 10L/min. The list of MRMs used and chromatographic conditions were extensively described previously [Huynh et al. 2019] |
| Ion Mode: | POSITIVE |
