Ab triple tof 5600 plus mass spectrometer

HPLC-MS analysis was conducted by AB Triple TOF 5600 plus Mass spectrometer (AB SCIEX, Framingham, USA) in positive electrospray ionization (ESI) mode with the liquid chromatography system (Waters Corp., Milford, MA, USA). Specific experimental methods and conditions were conducted as we reported previously [30 (link)]. The accurate mass and compounds’ structure was calculated by Peak View Software (AB SCIEX, version 1.2.0.3).

Cells grown in 10-cm tissue culture dishes were washed twice with PBS and lysed in 80% (vol/vol) methanol at −78 °C to extract intracellular metabolites. Cell debris was removed by centrifugation at 4 °C. The supernatant containing metabolites was dried under N2 and then were resuspended using 80 µL LC/MS-grade water. Samples were injected and analysed using AB Triple TOF 5600 plus mass spectrometer (AB/SCIEX, USA) coupled to an HPLC system (AB/SCIEX). ATP and GTP were measured as described in previous studies [17 (link), 38 (link)]. Peaks representing each metabolite were extracted and analysed using MultiQuant software.