Ultra-high-performance liquid chromatography high-resolution mass spectrometry analysis
We have previously carried out a gas chromatography-mass spectrometry (GC-MS)-based metabolite profiling analysis in leaf disks from ferns (M. scolopendria and P. aureum ) and angiosperms (V. unguiculata and N. tabacum ) species harvested throughout the day (05:00 h, 08:00 h, 14:00 h and 17:00 h) (Lima et al. 2019). This GC-MS platform is mostly related to the analysis of primary metabolites (Lisec et al. 2006). Here, we extended the metabolic characterization of ferns and angiosperms by carrying out an ultra-high-performance liquid chromatography high-resolution mass spectrometry (LC-MS) analysis, which mostly detect secondary metabolites (Perez de Souza et al. 2021), using samples from the same experiment carried out previously (Lima et al. 2019). Metabolites extraction and the LC-MS analysis were carried out exactly as described earlier (Tohge & Fernie 2010). The mass spectral data was first analysed using a metabolic fingerprinting approach followed by the identification of metabolites through the use of MS-DIAL software (Tsugawa et al. 2015), XCMS (Smith et al. 2006) and the MetaboanalystR package on R (Pang et al. 2020).