Workflow used for the UHPLC-PDA-CAD-ESI-HRMS 2 metabolite profiling and metabolite annotation of the Celastraceae set. a. Extract preparation: To obtain the crude extract, each sample underwent a three-step extraction process involving ethyl acetate maceration assisted by an ultrasound bath, followed by filtration and subsequent drying under nitrogen flux. b. SPE filtration: To remove highly lipophilic compounds, the crude extracts were dissolved in methanol and filtered through a C18-SPE cartridge. The filtrate was then dried under nitrogen flux to obtain an enriched extract. c. Sample preparation and UHPLC-MS 2 metabolite profiling: The enriched extracts were dissolved in DMSO to a concentration of 5 mg/mL and a 100 μL aliquot was transferred (without further dilution) to a 700 μL 96-well plate for UHPLC-PDA-CAD-HRMS 2 metabolite profiling. d. Data conversion and data processing: The .raw data files were converted to the .mzML open-source format. The converted data was analyzed using MZmine3 to generate a 'quantitative table' of features (peak with m/z@rt) present in the data set, as well as a spectral file (specs.mgf) that includes the MS 2 spectral information for each feature. e. Molecular Networking: Upon conversion, the .mzML files, along with the quantitative table and spectral .mgf files, were uploaded to the GNPS website. An FBMN was generated and an automated search for spectral matches against experimental DBs was performed. f. Spectral annotation with ISDB: The spectral specs.mgf file was subjected to a taxonomically informed metabolite annotation pipeline (ISDB annotation followed by taxonomical reweighting) using TimaR. g. Spectral annotation with Sirius: The specs_sirius.mgf file was processed with Sirius to obtain the chemical classes, molecular formulas, and structural annotations with CSI-FinguerID , and CANOPUS .