Ltq linear ion trap ms

The defatted methanol extract was analyzed using LC–MS–MS. A Shimadzu LC-10 HPLC with a Grace Vydac Everest Narrowbore C-18 column (100 mm × 2.1 mm i.d., 5 µm, 300 Å). LC–MS, connected to an LTQ Linear Ion Trap MS (Thermo Finnigan, San Jose, CA) was utilized with a mass range of 100–2000 m/z. A 2 µL sample was injected using an autosampler. A 35 min method was used as follows: 5 min isocratic run using 5% acetonitrile (Acn) and 0.05% formic acid (FA), then a gradient was run for 25 min until 95% AcN 0.05% FA. Finally, there was 5 min of conditioning the column with 5% AcN and 0.05% FA. The data were processed and analyzed using foundation 3.1_Xcalibur_3.1.6610. Furthermore, the raw data files were converted to mzXML format using MSConvert from the ProteoWizard suite [70 ]. The molecular network was created using the Global Natural Products Social Molecular Networking (GNPS) online workflow [28 (link),29 (link)]. The spectra in the network were then searched against the GNPS spectral libraries and published data.

For LC-MS/MS analysis of the extract, a Shimadzu LC-10 HPLC, equipped with a Grace Vydac Everest Narrow bore C-18 column [internal diameter (100 mm × 2.1 mm) and particle size 300 Å), was utilized. An LTQ Linear Ion Trap MS (Thermo Finnigan, San Jose, CA), with a mass range of 100–2,000 m/z, was utilized. A sample size of 2 µL was auto-injected. A gradient elution pattern (continued for 15 min) was employed using gradients of 5% CH₃CN and 0.05% HCOOH, until 95% CH₃CN 0.05% HCOOH. For data analysis and interpretation, the software MSDIAL ver.5.1.230912 and MZmine 3 were utilized. The files of the raw data were then converted to mzXML format using MSConvert from the ProteoWizard suite (Al Mousa et al., 2022 (link)).