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MB Sample ID: SA338093
Local Sample ID: | CD043_004 |
Subject ID: | SU003237 |
Subject Type: | Cultured cells |
Subject Species: | Mus musculus |
Taxonomy ID: | 10090 |
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Combined analysis:
Analysis ID | AN005114 |
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Analysis type | MS |
Chromatography type | HILIC |
Chromatography system | Vanquish UHPLC |
Column | Atlantis Premier BEH Z-HILIC (100 x 2.1 mm, 1.7 um) |
MS Type | ESI |
MS instrument type | Orbitrap |
MS instrument name | Thermo Fisher Scientific Orbitrap Exploris 240 |
Ion Mode | NEGATIVE |
Units | counts per second (cps) |
Chromatography:
Chromatography ID: | CH003870 |
Chromatography Summary: | Snap-frozen gastruloids were sent to EMBL-Heidelberg, Germany, for the metabolomics analysis. Reagents: LC-MS grade water, acetonitrile and methanol were obtained from Th. Geyer (Germany). High-purity ammonium acetate, ammonium hydroxide, and formic acid were purchased from Merck (Germany). Stable isotope labelled amino acids (MSK-MET1-1; Cambridge Isotope Laboratories, MA, USA) were used as internal standards for untargeted metabolomics. LC-MS/MS analysis was performed on a Vanquish UHPLC system coupled to an Orbitrap Exploris 240 high-resolution mass spectrometer (Thermo Fisher Scientific, MA, USA) in negative ESI (electrospray ionization) mode. Chromatographic separation was carried out on an Atlantis Premier BEH Z-HILIC column (Waters, MA, USA; 2.1?mm x 100 mm, 1.7 µm) at a flow rate of 0.25 mL/min. The mobile phase consisted of water:acetonitrile (9:1, v/v; mobile phase A) and acetonitrile:water (9:1, v/v; mobile phase B), which were modified with a total buffer concentration of 10 mM ammonium acetate. The aqueous portion of each mobile phase was adjusted to pH 9.0 via addition of ammonium hydroxide. The following gradient (20 min total run time including re-equilibration) was applied (time[min]/%B): 0/95, 2/95, 14.5/60, 16/60, 16.5/95, 20/95. Column temperature was maintained at 40°C, the autosampler was set to 4°C and sample injection volume was 7 µL. |
Methods Filename: | LC_MS_MS_Full_Protocol.pdf |
Instrument Name: | Vanquish UHPLC |
Column Name: | Atlantis Premier BEH Z-HILIC (100 x 2.1 mm, 1.7 um) |
Column Temperature: | 40 |
Flow Gradient: | time [min]/%B - 0/95, 2/95, 14.5/60, 16/60, 16.5/95, 20/95 |
Flow Rate: | 0.25mL/min |
Solvent A: | water:acetonitrile 9:1, v/v; 10mM ammonium acetate |
Solvent B: | acetonitrile:water 9:1, v/v; 10mM ammonium acetate |
Chromatography Type: | HILIC |