Feature-based molecular networking and MS2LDA analysis for the dereplication of adjacent bis-tetrahydrofuran...

Annonaceous acetogenins are a group of natural polyketide compounds possessing notable cytotoxic and antitumor properties. Mass spectrometry (MS) techniques can be used for the structural determination of these compounds, including the location of functional groups along the long alkyl chain.

This study aims to develop a convenient liquid chromatography (LC)-MS-based method for the dereplication of acetogenins in plant extracts using a molecular networking approach.

The LC-electrospray ionization (ESI)-MS/MS spectra of pure adjacent bis-tetrahydrofuran (THF) acetogenins isolated from Uvaria rufa (Annonaceae) were acquired, along with those of the crude ethyl acetate and hexanes fractions of the plant extract, followed by dereplication and molecular networking analysis using the Global Natural Products Social Molecular Networking (GNPS) platform.

A high level of fragmentation of the protonated molecules [M + H]⁺ was observed at collision energies of 37.5 and 25.0 eV. The application of feature-based molecular networking (FBMN) allowed for distinguishing diastereoisomers based on different retention times in the reversed-phase high-performance liquid chromatography method. The acetogenin possessing one or more additional OH groups on the methyl-terminal chain side of the OH-flanked bis-THF ring unit were grouped separately from those lacking such substructure. Furthermore, the MS2LDA analysis revealed shared Mass2Motifs among acetogenins, confirming the structural relations within the molecular network.

The ESI-MS/MS-based molecular networking method provided an effective strategy for the dereplication of acetogenins in plant extracts. It is anticipated that this molecular networking approach could be extended to other types of acetogenins to facilitate rapid identification of this class of compounds.