{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,2]],"date-time":"2026-10-02T19:54:30Z","timestamp":1790970870128,"version":"4.1.0"},"reference-count":53,"publisher":"MDPI AG","issue":"20","license":[{"start":{"date-parts":[[2025,10,20]],"date-time":"2025-10-20T00:00:00Z","timestamp":1760918400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100012166","name":"National Key R&D Program of China","doi-asserted-by":"publisher","award":["2022YFE0108300"],"award-info":[{"award-number":["2022YFE0108300"]}],"id":[{"id":"10.13039\/501100012166","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Basic Agricultural Research Project of Wenzhou","award":["N20220013"],"award-info":[{"award-number":["N20220013"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Plants"],"abstract":"<jats:p>Background: Broccoli contains diverse phytochemicals, including glucosinolates and their hydrolysis products, with potential nutritional and bioactive properties. Accurate metabolite profiling requires optimized sample preparation and comprehensive databases. Methods: A rapid enzymatic deactivation method with 70% methanol, implemented prior to cryogrinding, was evaluated for processing freeze-dried and fresh broccoli florets, which were compared as plant materials. A widely targeted, organ-resolved metabolite database was constructed by integrating over 612 reported phytochemicals with glucosinolate degradation products. LC-HRMS combined with MS-DIAL and GNPS was employed for metabolite detection and annotation. Results: Freeze-dried samples yielded nearly twice the number of glucosinolates, isothiocyanates, and nitriles compared with standard-processed fresh tissue. Methanol pre-treatment preserved metabolite integrity in fresh samples, achieving comparable sensitivity to freeze-dried material. Using the integrated database, 998 metabolites were identified or tentatively characterized, including amino acids, carboxylic acids, phenolics, alkaloids, terpenoids, and glucosinolate derivatives. Cross-platform reproducibility was improved and false positives reduced. Conclusions: Optimized sample preparation combined with a curated metabolite database enables high-confidence, comprehensive profiling of broccoli florets phytochemicals. The resulting dataset provides a valuable reference for studies on genotype\u2013environment interactions, nutritional quality, and functional bioactivity of cruciferous vegetables.<\/jats:p>","DOI":"10.3390\/plants14203223","type":"journal-article","created":{"date-parts":[[2025,10,20]],"date-time":"2025-10-20T13:54:41Z","timestamp":1760968481000},"page":"3223","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["An Integrated Approach for the Comprehensive Characterization of Metabolites in Broccoli (Brassica oleracea var. Italica) by Liquid Chromatography High-Resolution Tandem Mass Spectrometry"],"prefix":"10.3390","volume":"14","author":[{"given":"Zhiwei","family":"Hu","sequence":"first","affiliation":[{"name":"National and Local Joint Engineering Research Center of Ecological Treatment Technology for Urban Water Pollution, Zhejiang Provincial Key Laboratory of Water Ecological Environment Treatment and Resource Protection, College of Life and Environmental Science, Wenzhou University, Wenzhou 325035, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Meijia","family":"Yan","sequence":"additional","affiliation":[{"name":"National and Local Joint Engineering Research Center of Ecological Treatment Technology for Urban Water Pollution, Zhejiang Provincial Key Laboratory of Water Ecological Environment Treatment and Resource Protection, College of Life and Environmental Science, Wenzhou University, Wenzhou 325035, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chenxue","family":"Song","sequence":"additional","affiliation":[{"name":"National and Local Joint Engineering Research Center of Ecological Treatment Technology for Urban Water Pollution, Zhejiang Provincial Key Laboratory of Water Ecological Environment Treatment and Resource Protection, College of Life and Environmental Science, Wenzhou University, Wenzhou 325035, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Muneo","family":"Sato","sequence":"additional","affiliation":[{"name":"RIKEN Center for Sustainable Resource Science, Yokohama 230-0045, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shiwen","family":"Su","sequence":"additional","affiliation":[{"name":"Southern Zhejiang Key Laboratory of Crop Breeding, Wenzhou Academy of Agricultural Sciences, Wenzhou Vocational College of Science and Technology, Wenzhou 325006, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sue","family":"Lin","sequence":"additional","affiliation":[{"name":"National and Local Joint Engineering Research Center of Ecological Treatment Technology for Urban Water Pollution, Zhejiang Provincial Key Laboratory of Water Ecological Environment Treatment and Resource Protection, College of Life and Environmental Science, Wenzhou University, Wenzhou 325035, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Junliang","family":"Li","sequence":"additional","affiliation":[{"name":"National and Local Joint Engineering Research Center of Ecological Treatment Technology for Urban Water Pollution, Zhejiang Provincial Key Laboratory of Water Ecological Environment Treatment and Resource Protection, College of Life and Environmental Science, Wenzhou University, Wenzhou 325035, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8141-8059","authenticated-orcid":false,"given":"Huixi","family":"Zou","sequence":"additional","affiliation":[{"name":"National and Local Joint Engineering Research Center of Ecological Treatment Technology for Urban Water Pollution, Zhejiang Provincial Key Laboratory of Water Ecological Environment Treatment and Resource Protection, College of Life and Environmental Science, Wenzhou University, Wenzhou 325035, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zheng","family":"Tang","sequence":"additional","affiliation":[{"name":"Southern Zhejiang Key Laboratory of Crop Breeding, Wenzhou Academy of Agricultural Sciences, Wenzhou Vocational College of Science and Technology, Wenzhou 325006, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0802-6208","authenticated-orcid":false,"given":"Masami Yokota","family":"Hirai","sequence":"additional","affiliation":[{"name":"RIKEN Center for Sustainable Resource Science, Yokohama 230-0045, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5127-494X","authenticated-orcid":false,"given":"Xiufeng","family":"Yan","sequence":"additional","affiliation":[{"name":"National and Local Joint Engineering Research Center of Ecological Treatment Technology for Urban Water Pollution, Zhejiang Provincial Key Laboratory of Water Ecological Environment Treatment and Resource Protection, College of Life and Environmental Science, Wenzhou University, Wenzhou 325035, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,10,20]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1047","DOI":"10.1016\/j.tplants.2018.09.011","article-title":"Contribution of untargeted metabolomics for future assessment of biotech crops","volume":"23","author":"Christ","year":"2018","journal-title":"Trends Plant Sci."},{"key":"ref_2","doi-asserted-by":"crossref","unstructured":"Dabbousy, R., Rima, M., Roufayel, R., Rahal, M., Legros, C., Sabatier, J.-M., and Fajloun, Z. 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