Orbitrap id x

Non-polar extracts were reconstituted in 75 µl of IPA containing isotopically labeled lipid standards (a detailed list of standards is included in the SI) and analyzed by LC-MS using a ThermoFisher Scientific Accucore C30 150 × 2.1mm, 2.6 µm column paired with a Thermo Fisher Orbitrap ID-X in positive and negative polarity. Polar (80:20 CH₃OH:H₂O) extracts were reconstituted in 75 µl of 80:20 CH₃OH:H₂O containing isotopically labeled arginine, hypoxanthine, hippuric acid, and methionine (Cambridge Isotope Laboratories, Inc.) and analyzed by LC-MS using a Waters BEH Amide 150 × 2.1 mm, 1.7 µm column paired with a Thermo Fisher Orbitrap ID-X in positive and negative polarity. LC-MS/MS data for each mode of analysis was collected using three rounds of iterative DDA (Thermo Scientific AcquireX) performed on pooled test samples.
Data for each sample were collected in full MS1 with a resolution of 240,000 FWHM (full-width half-maximum) and MS/MS spectra of pooled samples were collected at a resolution of 30,000 FWHM using a 0.8 Da isolation window and stepped HCD collision energies of 15, 30, and 45.

All NMR spectroscopic
data (1D and 2D) were acquired on an 18.8 T (800 MHz for ¹H and 201 MHz for ¹³C) Bruker Avance IIIHD instrument
equipped with a 3 mm triple resonance cryoprobe or a 16.4 T (700 MHz
for ¹H and 175 MHz for ¹³C) Bruker Avance IIIHD
instrument with a 5 mm indirect broadband cryoprobe. CDCl₃ or pyridine-d₅ was used as the NMR solvent
for all samples and referenced to residual solvent peaks δ_H 7.26, δ_C 77.2 for CDCl₃ and δ_H 7.22, δ_C 123.9 for pyridine-d₅. NMR spectroscopic data were processed and analyzed
using MestReNova 11.0.4.
High-resolution mass spectrometric
data were acquired in a Thermo Scientific Orbitrap ID-X instrument.
Both normal and reversed phase HPLC chromatographic separations were
performed using a Waters 1525 binary pump coupled with a Waters 2996
photodiode array detector and an Altech 800 evaporative light scattering
detector.