Reverse-phase (RP) ultra-high
performance liquid chromatography–mass spectrometry (UHPLC–MS)
analysis was performed with a Thermo Accucore C30, 150 × 2.1
mm2, 2.6 μm particle size column mounted in a Vanquish
LC coupled to an Orbitrap ID-X Tribrid mass spectrometer (ThermoFisher
Scientific). The mobile phases and chromatographic gradients used
are described inTable S6 . MS data were
acquired in positive and negative ion modes in the 150–2000 m/z range with a 120,000 mass resolution
setting. The most relevant MS parameters are provided inTable S7 . Samples were kept at 4 °C in the
autosampler during LC–MS analysis, while the column temperature
was set to 50 °C. An injection volume of 2 μL was used
for all runs. For lipid annotation, MS/MS experiments were performed
using the Thermo Scientific AcquireX data acquisition workflow. Tandem
MS data were acquired at a resolution of 30,000 and an isolation window
of 0.4 m/z. Precursor ions were
fragmented with HCD and CID activation methods. For HCD, stepped normalized
collision energy (NCE) of 15, 30, and 45 and a CID collision energy
of 40 were used to fragment the precursor ions.
performance liquid chromatography–mass spectrometry (UHPLC–MS)
analysis was performed with a Thermo Accucore C30, 150 × 2.1
mm2, 2.6 μm particle size column mounted in a Vanquish
LC coupled to an Orbitrap ID-X Tribrid mass spectrometer (ThermoFisher
Scientific). The mobile phases and chromatographic gradients used
are described in
acquired in positive and negative ion modes in the 150–2000 m/z range with a 120,000 mass resolution
setting. The most relevant MS parameters are provided in
autosampler during LC–MS analysis, while the column temperature
was set to 50 °C. An injection volume of 2 μL was used
for all runs. For lipid annotation, MS/MS experiments were performed
using the Thermo Scientific AcquireX data acquisition workflow. Tandem
MS data were acquired at a resolution of 30,000 and an isolation window
of 0.4 m/z. Precursor ions were
fragmented with HCD and CID activation methods. For HCD, stepped normalized
collision energy (NCE) of 15, 30, and 45 and a CID collision energy
of 40 were used to fragment the precursor ions.