Chromatographic separation of plasma was performed with Vanquish ultra high performance liquid chromatograph (UHPLC) system (Thermo, USA). An Accucore HILIC column (2.1 × 150 mm, 2.6 µm) was used in the study. The column was maintained at 40 °C, and the ow rate was 0.3 mL/min. The mobile phase for the positive ion mode was consisted of A (0.1% formic acid, 95% acetonitrile, 10 mM ammonium acetate) and B (0.1% formic acid, 50% acetonitrile, 10 mM ammonium acetate); the negative ion mode was consisted of A (95% acetonitrile, 10 mM ammonium acetate) and B (50% acetonitrile, 10 mM ammonium acetate). The process of linear gradient elution was shown as follows: 0-1 min, 98% A; 1-17 min, 98 - 50% A; 17-17.5 min, 50% A; 17.5-18 min, 50-98% A; 18-20 min, 98% A.
Mass spectrometer used in this study was Q Exactive™ HF-X (Thermo, USA) equipped with an electrospray ionization source (ESI) operating in positive (ESI+) and negative ion modes (ESI-). The MS main properties were set as follows: spray voltage, 3.2 KV; sheath gas ow rate, 35 arb; aux gas ow rate, 10 arb; scan range, m/z of 100-1500; capillary temperature, 320 °C. MS/MS data was acquired in data dependent acquisition (DDA) mode.
Mass spectrometer used in this study was Q Exactive™ HF-X (Thermo, USA) equipped with an electrospray ionization source (ESI) operating in positive (ESI+) and negative ion modes (ESI-). The MS main properties were set as follows: spray voltage, 3.2 KV; sheath gas ow rate, 35 arb; aux gas ow rate, 10 arb; scan range, m/z of 100-1500; capillary temperature, 320 °C. MS/MS data was acquired in data dependent acquisition (DDA) mode.