UltiMate 3000 HPLC (Dionex) with an XBridge Amide column [100 mm × 2.1 mm inner diameter (i.d.), 3.5 μm; Waters] was coupled to a Q Exactive (QE) mass spectrometer (Thermo Fisher Scientific) for metabolite separation and detection at room temperature. The mobile phase A reagent was composed of 20 mM ammonium acetate and 15 mM ammonium hydroxide in 3% acetonitrile in high-performance liquid chromatography (HPLC)–grade water (pH 9.0), while the mobile phase B reagent was acetonitrile. All solvents were of LC-MS grade, purchased from Thermo Fisher Scientific. The flow rate used was 0.15 ml/min from 0 to 10 min and 15 to 20 min, and 0.3 ml/min from 10.5 to 14.5 min. The linear gradient was as follows: 0 min 85% B, 1.5 min 85% B, 5.5 min 35% B, 10 min 35% B, 10.5 min 25% B, 14.5 min 35% B, 15 min 85% B, and 20 min 85% B.

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