LC-MS was performed with a Thermo Fisher Scientific LTQ XL mass spectrometer coupled with a Thermo Scientific UltiMate 3000 HPLC system. The MS detector was set with a +3.5 kV spray voltage, a 275 °C capillary temperature, a 300 °C heater temperature, a sheath gas flow of 25 L/min, auxiliary gas flow of 10 L/min, and a mass scan range of 80–900 m/z.
The gradient mobile phases were (A) 10 mM ammonium formate in 95:5 acetonitrile/water + 0.1% formic acid and (B) 10 mM ammonium formate in 50:50 acetonitrile/water + 0.1% formic acid for 30 min at a flow rate of 300 µL/min. Aliquots of serum extract (5 μL) were injected into a Thermo Fisher Scientific Accurore C18 Column (100 × 2.1 mm, 2.6 μm particle size), with a column temperature of 40 °C. MS data acquisition was performed using Thermo Fisher Scientific Xcalibur™ software v 2.2 SP1.48. Metabolites were determined through comparison of the ion features in the experimental samples to those of control samples based on retention time and molecular weight (m/z) using Thermo-Fisher Scientific Mass Frontier 7.0 software [30 (link),31 ].
The gradient mobile phases were (A) 10 mM ammonium formate in 95:5 acetonitrile/water + 0.1% formic acid and (B) 10 mM ammonium formate in 50:50 acetonitrile/water + 0.1% formic acid for 30 min at a flow rate of 300 µL/min. Aliquots of serum extract (5 μL) were injected into a Thermo Fisher Scientific Accurore C18 Column (100 × 2.1 mm, 2.6 μm particle size), with a column temperature of 40 °C. MS data acquisition was performed using Thermo Fisher Scientific Xcalibur™ software v 2.2 SP1.48. Metabolites were determined through comparison of the ion features in the experimental samples to those of control samples based on retention time and molecular weight (m/z) using Thermo-Fisher Scientific Mass Frontier 7.0 software [30 (link),31 ].
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