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MB Sample ID: SA217829
| Local Sample ID: | SPT_C133W_1h_01311114 |
| Subject ID: | SU002360 |
| Subject Type: | Cultured cells |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
| Cell Biosource Or Supplier: | ATCC, CCL-247 |
| Cell Strain Details: | HCT116 |
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Combined analysis:
| Analysis ID | AN003716 |
|---|---|
| Analysis type | MS |
| Chromatography type | Reversed phase |
| Chromatography system | Thermo Vanquish |
| Column | Kinetex C8 (150 x 3mm,2.6um,100Å) |
| MS Type | ESI |
| MS instrument type | Orbitrap |
| MS instrument name | Thermo Q Exactive Orbitrap |
| Ion Mode | POSITIVE |
| Units | ion counts |
MS:
| MS ID: | MS003465 |
| Analysis ID: | AN003716 |
| Instrument Name: | Thermo Q Exactive Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | To quantify labeling on SL and deoxySL species from [U-13C16]palmitate a Q Exactive orbitrap mass spectrometer with a Vanquish Flex Binary UHPLC system (Thermo Scientific) was used with a Kinetex 2.6 μM C8 100 Å 150 x 3 mm LC column (Phenomenex) at 40°C. 5 μL of sample was injected. Chromatography was performed using a gradient of 2 mM ammonium formate and 0.2 % formic acid (mobile phase A) and 1 mM ammonium formate and 0.2 % formic acid in methanol (mobile phase B), at a flow rate of 0.5 mL/min. The LC gradient held at 82% B for 0-3 min, then ran from 82%-90% B in 3-4 min, then 90-99% in 4-18 min, held at 99% B for 7 min, then reduced from 99%-82% from 25-27 min, then held at 82% for a further 13 mins. Lipids were analyzed in positive mode using spray voltage 3 kV. Sweep gas flow was 5 arbitrary units, auxiliary gas flow 7 arbitrary units and sheath gas flow 50 arbitrary units, with a capillary temperature of 300°C. Full MS (scan range 150-2000 m/z) was used at 70 000 resolution with 1e6 automatic gain control and a maximum injection time of 200 ms. Data dependent MS2 (Top 6) mode at 17 500 resolution with automatic gain control set at 1e5 with a maximum injection time of 50 ms was used for peak identification, combined with known standards where possible. |
| Ion Mode: | POSITIVE |
