Feature-based Molecular Networking in phyllobilin research: New discoveries in Salvia officinalis

Phyllobilins are metabolites of chlorophyll formed during plant senescence, fruit ripening, and post-harvest storage of plant-derived foods. Though discovered over 30 years ago and widespread in nature, phyllobilins have only recently gained attention as a class of bioactive natural products with reported antioxidative, anti-inflammatory, and anti-proliferative properties [ 1 ]. They are typically identified based on their characteristic UV absorption spectra and fragmentation in tandem-mass spectrometry experiments [ 2 ]; however, MS 2 spectra are still absent from public natural products databases. This study presents a bioactivity-guided workflow integrating UHPLC-HRMS/MS and feature-based molecular networking (FBMN) to identify phyllobilins and structurally related metabolites. Extracts from Echinacea purpurea – a plant known to contain six structurally different phyllobilins – and Salvia officinalis , previously unexplored for phyllobilins, were fractionated and analyzed. Using the GNPSFBMN workflow, a molecular network was generated. To establish a reference framework, validated MS 2 spectra of phyllobilins from Echinacea were uploaded to GNPS, enabling annotation of potential phyllobilins in Salvia officinalis . These candidate compounds were subsequently isolated and their structures were confirmed via 1D and 2D NMR. By contributing high-quality phyllobilin MS 2 spectra to public databases and demonstrating a robust analytical approach, this work facilitates the discovery of novel phyllobilins in crude and complex plant extracts and expands the chemical space of chlorophyll metabolites supporting future studies of their bioactivity and health promoting potential. Publication History Article published online: 05 March 2026 © 2026. Thieme. All rights reserved. Georg Thieme Verlag KG Oswald-Hesse-Straße 50, 70469 Stuttgart, Germany