Everest narrowbore c18 column

Extracts of bee products were analyzed using LC–MS–MS. A Shimadzu LC-10 HPLC instrument with a Grace Vydac Everest Narrowbore C₁₈ column (100 mm × 2.1 mm i.d., 5 µm, 300 Å). LC-MS connected to an LCQ electrospray ion trap MS (Thermo Finnigan, San Jose, CA, USA) was utilized with a mass range of 200–5000 m/z. A 2 µL sample was injected using an autosampler. The solvents used were 95% H₂O in formic acid (0.1%) (A) and 95% ACN in formic acid (0.1%) (B). Gradient elution ranged from 5% to 95% (B), followed by column conditioning to 5% (B) at a 300 µL/min flow rate. The elution time was 60 min.
Foundation 3.1 Xcalibur 3.1.6610 was used to analyze the data. Additionally, MS Convert from the ProteoWizard suite (https://proteowizard.sourceforge.io/download.html; accessed on 8 November 2022) was used to convert the raw data files to mzXML format. The Global Natural Products Social Molecular Networking (GNPS) online workflow was used to generate the molecular network [3 (link),4 (link)]. The network’s spectra were then validated against the spectral libraries and literature data of GNPS.
Cytoscape software was used to analyze and edit the molecular networks. The parent mass of each node served as a label. A pie slice proportional to the number of MS/MS spectra for each parent mass was established, with colors designating the sources of the samples.

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)).