Phytochemical characterization and valorization of Symphytum officinale L. roots from continental Croatia

Comfrey ( Symphytum officinale L., Boraginaceae) root-based preparations have been used for the management of pain and inflammation associated with musculoskeletal disorders, including conditions affecting joints, bones, and muscles [ 1 ]. Metabolomic analyses have shown the complex phytochemical profile of comfrey roots, comprising polysaccharides, allantoin, cinnamic acid derivatives, and pyrrolizidine alkaloids. Furthermore, pharmacological studies have revealed the mechanistic basis of their anti-inflammatory, analgesic, wound-healing, anti-irritant, and osteo-regenerative activities. Key bioactive constituents, such as allantoin, rosmarinic acid, globoidnans A and B, rabdosiin, have been identified as principal contributors to the therapeutic efficacy of Symphytum-derived formulations, particularly in mediating anti-inflammatory and wound-healing responses [ 2 ]. The objective of this study was to evaluate the phytochemical composition of comfrey root samples collected from continental Croatia, with the aim of exploring their potential as sources of bioactive compounds for the development of formulations intended for both human and veterinary applications. Root samples were obtained from three distinct geographical regions, Zagreb County, Baranja County, and Požega-Slavonia County. The plant material was extracted using 65% ethanol through maceration, employing a drug-extract ratio (DER) of 1:10 (w/v). The phytochemical profiling of comfrey extracts was conducted using liquid chromatography-high resolution tandem mass spectrometry (LC-HRMS/MS). All analyzed extracts displayed similar chemical compositions, characterized by the presence of ten phenolic acids (including caffeic acid, rosmarinic acid, globoidnans A and B, rabdosiin), seven pyrrolizidine alkaloids (e.g., intermedine/lycopsamine, acetylintemedine/acetyllycopsamine, symphytine, and their corresponding N-oxides), and three organic acids. Accordingly, this study highlights S. officinale roots collected from various regions of Croatia as a valuable source of bioactive compounds, with potential applicability in the development of topical formulations for both human and veterinary use. Publication History Article published online: 05 March 2026 © 2026. Thieme. All rights reserved. Georg Thieme Verlag KG Oswald-Hesse-Straße 50, 70469 Stuttgart, Germany