Propose <p> Aerial parts from the herb <i>Anacyclus valentinus</i> L. are traditionally used by the local populations in Algeria to treat infections and several diseases such as diabetes, cholesterol disorders, abdominal pains, indigestion, digestive disorders, colic and pains, and commonly employed in culinary preparations. They are rich in bioactive compounds, such as terpenes and flavonoids. This work aimed to explore <i>A. valentinus</i> as a potential source of bioactive molecules with food and pharmaceutical applications.</p> Methods <p>Ethanol crude extract and fractions (hexane, chloroform, ethyl acetate, <i>n</i>-butanol and aqueous) were obtained from aerial parts of this herb and evaluated for radical scavenging activity (RSA) on 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2′-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) and OH radicals, ferric reducing antioxidant power (FRAP), silver ions (Ag⁺) reducing potential, and for ferric-ion-based total antioxidant capacity. Extracts were also studied for in vitro inhibition of enzymes implicated on the onset of neurological diseases (butyrylcholinesterase: BuChE) and type-2 diabetes mellitus (T2DM, α-amylase). The total contents of phenolics (TPC), flavonoids (TFC), and tannins (TTC) were determined by colorimetric methods. Qualitative analyses of this herb were performed by ultra-high performance liquid chromatography system coupled with electrospray ionization tandem mass spectrometry (UHPLC-ESI/MSMS).</p> Results <p>The results revealed that the polar extracts had significant RSA and capacity to reduce iron, and silver ions (Ag⁺). The extracts had a higher activity towards BuChE and α-amylase, especially the hexane fraction and crude ethanol extract. The <i>n</i>-butanol fraction had the highest TPC (214.64&#xa0;mg GAE/g DE), ethyl acetate had the highest flavonoid levels (145.44 CE/g DE), whereas the ethanol crude extract had the highest TTC (402.38&#xa0;mg CE/g DE). Isorhamnetin (1), scilliglaucoside (2), butein (3), gallocatechin (4), protocatechuic acid hexoside (5), myricetin derivative (6), ferulic acid derivative (7), quercetin (8), luteolin 3-<i>O</i>-glucoside (9), and caffeic acid derivative (10) were identified in ethyl acetate and <i>n</i>-butanol fractions.</p> Conclusion <p>The findings suggest <i>A. valentinus</i> as a promising source of biologically active molecules with food, pharmaceutical, and biomedical applications.</p> Graphical Abstract <p></p>