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Charting the Undiscovered Metabolome with Synthetic Multiplexing.

Title: Charting the Undiscovered Metabolome with Synthetic Multiplexing.
Authors: Patan A; Xing S; Charron-Lamoureux V; Hu Z; Deleray V; Agongo J; El Abiead Y; Mannochio-Russo H; Mohanty I; Gouda H; Zemlin J; Rajkumar P; Lee C; Leanos D; Weimann N; Tsuda W; Giddings S; Bui T; Kvitne KE; Zhao HN; Zuffa S; Nguyen V; Andrade A; Gonçalves Nunes WD; Caraballo-Rodríguez AM; Caetano David L; Carver J; Bandeira N; Wang M; Burnett LA; Siegel D; Dorrestein PC
Source: BioRxiv : the preprint server for biology [bioRxiv] 2025 Nov 19. Date of Electronic Publication: 2025 Nov 19.
Publication Type: Journal Article; Preprint
Language: English
Journal Info: Country of Publication: United States NLM ID: 101680187 Publication Model: Electronic Cited Medium: Internet ISSN: 2692-8205 (Electronic) Linking ISSN: 26928205 NLM ISO Abbreviation: bioRxiv Subsets: PubMed not MEDLINE
Abstract: In untargeted metabolomics, reference MS/MS libraries are essential for structural annotation, yet currently explain only 6.9% of the more than 1.7 billion MS/MS spectra in public repositories. We hypothesized that many unannotated features arise from simple, biologically plausible transformations of endogenous and exposure-derived compounds. To test this, we created a reference resource by synthesizing over 100,000 compounds using multiplexed reactions that mimic such biochemical transformations. 91% of the compounds synthesized are absent from existing structural databases. Through improvements in the construction of the computational infrastructure that enables pan repository-scale MS/MS comparisons, searching this biologically inspired MS/MS library increased the overall reference-based match rate by 17.4%, yielding over 60 million new matches and raising the global pan-repository MS/MS annotation rate to 8.1%. By facilitating structural hypotheses for previously uncharacterized MS/MS data, this framework expands the accessible detectable biochemical landscape across human, animal, plant, and microbial systems, revealing previously undescribed metabolites such as ibuprofen-carnitine and 5-ASA-phenylpropionic acid conjugates arising from drug-host and host-microbiome co-metabolism.
Entry Date(s): Date Created: 20251203 Date Completed: 20251203 Latest Revision: 20251203
Update Code: 20260130
PubMed Central ID: PMC12667992
DOI: 10.1101/2025.11.18.689170
PMID: 41332743
Database: MEDLINE

Journal Article; Preprint