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Due to its increased bioavailability and lower toxicity, the green biosynthesis of metal nanoparticles (M-NPs) utilizing plant extract as a capping agent has been demonstrated over a number of years. Thus, the current study sought to examine the <jats:italic>in vitro<\/jats:italic> biological activity of copper oxide nanoparticles\u00a0 (CuO-NPs) and selenium nanoparticles (Se-NPs) that were biosynthesized using aqueous, methanolic, and petroleum ether extracts from <jats:italic>S. alba<\/jats:italic> seeds. Phytochemical and <jats:italic>in vitro <\/jats:italic>biological activities (antioxidant, scavenging, anti-diabetic, anti-acetylcholinesterase, anti-arthritic, anti-inflammatory, and cytotoxic activities) were assayed in all prepared extracts before and after being used for the biosynthesis of the M-NPs. It was found that the total methanolic extract possessed the highest biological activities compared to other native extracts. The LC-ESI-MS\/MS analysis of secondary metabolites showed that the total methanolic extract contained 7 phenolic acids and 9 flavonoid aglycones. This helped find the active ingredients. We characterized 8 phenolic acid derivatives, 7 flavonoid glycosides, 4 aliphatic glucosinolates, and 3 aromatic aryl glucosinolates in the aqueous extract. Furthermore, the methanolic extract contains the highest concentrations of total polyphenols, condensed tannins, and total flavonoid compounds. The biosynthesized Se-NPs using methanolic extract showed higher <jats:italic>in vitro<\/jats:italic> biological activities compared to those of the biosynthesized CuO-NPs. The median lethal dose (LD<jats:sub>50<\/jats:sub>) showed that the biosynthesized Se-NPs using the studied extracts appeared safer compared to those of the biosynthesized CuO-NPs. The findings of this study concluded that the total methanolic extract is the most suitable bioresource for biosynthesizing Se-NPs through green nanotechnology, with higher biological efficiency in relation to its metabolite fingerprint.<\/jats:p>","DOI":"10.1038\/s41598-025-95347-6","type":"journal-article","created":{"date-parts":[[2025,4,21]],"date-time":"2025-04-21T16:08:16Z","timestamp":1745251696000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":15,"title":["Phytochemical analysis and biological study on Sinapis alba L. seeds extract incorporated with metal nanoparticles, in vitro approach"],"prefix":"10.1038","volume":"15","author":[{"given":"Wael Mahmoud","family":"Aboulthana","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Amal M.","family":"El-Feky","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Noha El-Sayed","family":"Ibrahim","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ahmed A. 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