Chromatographic separation was accomplished by hydrophobic interaction liquid chromatography using a Luna NH2 3 μm, 2 mm × 100 mm column, (Phenomenex) on a Vanquish Flex Quaternary UHPLC system (Thermo Fisher Scientific). Mobile phase A consisted of 10 mM NH4HCO3, 2% NH4OH (Thermo Fisher Scientific). Mobile phase B consisted of acetonitrile (Honeywell Burdick & Jackson).

MS analyses were performed on an Orbitrap Fusion Lumos Tribrid mass spectrometer (Thermo Fisher Scientific) interfaced with an H-ESI electrospray source (Thermo Fisher Scientific). General instrumental conditions were RF 30%; sheath gas, auxiliary gas, and sweep gas of 35, 11, and 0 arbitrary units, respectively; ion transfer tube temperature of 300°C; vaporizer temperature of 250°C; and spray voltage of 2500 V for positive mode and 3500 V for negative mode. Data were collected for each sample in negative mode, using 2 different mass ranges (70–700 m/z and 220–900 m/z) to enhance sensitivity for larger, less-abundant compounds, and in positive mode (70–900 m/z) (74, 75).

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