Non-targeted metabolite profiling reveals biochemical diversity of three East Asian mosses

Mosses are known for their rich diversity in bioactive compounds and their essential role in maintaining ecosystems. While genetic and metabolomics studies have been conducted on some model species, research on comprehensive metabolomics of common mosses in East Asia remains limited. In this study, we conducted an untargeted analysis of three East Asian moss species, Niphotrichum japonicum, Calohypnum plumiforme, and Polytrichum formosum, by ultrasonic-assisted extraction and liquid chromatography coupled with high-resolution mass spectrometry. This approach facilitated the detailed profiling and compound classification through reference databases. Tandem mass spectrometry with positive mode tentatively identified 188, 183, and 186 metabolites, respectively, with lipids and terpenoids as the predominant class. Among these, 25 potential biomarkers were identified to differentiate between species. Multivariate analysis, including principal component analysis, revealed distinct metabolic profiles for each species, representing taxonomical differences, confirming data reproducibility, and clear species differentiation. Enrichment analysis further highlighted the upregulation of unsaturated fatty acid biosynthesis across all moss species. This is the first report utilizing an untargeted metabolomics approach with chemometrics to differentiate moss species cultivated in the same location, offering a new foundation for studying the metabolic responses of East Asian mosses.