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).

Note: The content above has been extracted from a research article, so it may not display correctly.

Please log in to submit your questions online.
Your question will be posted on the Bio-101 website. We will send your questions to the authors of this protocol and Bio-protocol community members who are experienced with this method. you will be informed using the email address associated with your Bio-protocol account.

We use cookies on this site to enhance your user experience. By using our website, you are agreeing to allow the storage of cookies on your computer.