Exactive plus benchtop orbitrap mass spectrometer

Chromatographic analysis was performed with the DIONEX Ultimate 3000 HPLC system coupled to a chromatographic column (Phenomenex Synergi 4 u Hydro-RP 80A 250 × 3.0 mm) set at 40 °C and a flow rate of 0.9 mL/min. Gradients of mobile phases (mobile phase A: 0.1% formic acid in water and mobile phase B: 0.1% formic acid in acetonitrile) were performed over a total of 25 min. MS analysis was carried out on a Thermo Scientific Exactive Plus Benchtop Orbitrap mass spectrometer. The heated electrospray ionization source (HESI II) was used in positive and negative ion modes. The instrument was operated in full scan mode from m/z 67 to m/z 1000. High-resolution accurate mass (HRAM) full-scan MS and top five MS/MS spectra were collected in a data-dependent fashion at a resolving power of 70,000 and 35,000 at FWHM m/z 200, respectively. The post-treatment of data was performed using the MZmine2 version 2.53 (http://mzmine.github.io/; Boston, MA, USA) [45 (link)]. Metabolites were identified using the Human Metabolome Database version 4.0 [46 (link)]. LC-MS analyses were performed in a blinded assessment; that is, without revealing the identity of the samples to the mass spectroscopist prior to analyses.

Chromatographic analysis was performed with the DIONEX Ultimate 3000 HPLC system coupled to a chromatographic column (Phenomenex Synergi 4 u Hydro-RP 80A 250_3.0 mm) set at 40 °C and a flow rate of 0.9 mL/min. Gradients of mobile phases (mobile phase A: 0.1% formic acid in water and mobile phase B: 0.1% formic acid in acetonitrile) were performed over a total of 25 min. MS analysis was carried out on a Thermo Scientific Exactive Plus Benchtop Orbitrap mass spectrometer. The heated electrospray ionization source (HESI II) was used in positive and negative ion modes. The instrument was operated in full scan mode from m/z 67 to m/z 1000. Post-treatment of data was performed using the MZmine2 version 2.53 (http://mzmine.github.io/) [22 (link)]. Metabolites were identified using the Metlin database (https://metlin.scripps.edu/).