UHPLC-MS/MS Analysis of Metabolites

The samples were prepared using a method described by Oliveira et al. [56 (link)] The crude methanol extract was separated using a C18 Reversed-phase Hypersil GOLD aQ column (100 × 2.1 mm ~1.9 µm) (Thermo, Waltham, MA, USA) at 30 °C on a Dionex Ultimate 3000 UHPLC with a diode-array DAD-3000 detector (Thermo Fisher Scientific, Waltham, MA, USA). The UHPLC-MS/MS analyses were conducted based on a method described by Buzgaia et al. [4 ]. The MS data analyses were conducted using the ThermoXcalibur 2.2 SP1.48 software (Thermo Fisher Inc. Waltham, MA, USA) and based on readily available literature data. The MS data were firstly converted into the mzXML format using the MSConvert software. The generated spectral information was then uploaded to MZmine-2.5.3-Windows to generate the MS/MS data [57 (link),58 (link)].