Phytochemical content, antioxidant, antimicrobial, antibiofilm, and enzyme inhibition activities of different extracts of Hyoscyamus aureus L. (Solanaceae) species

In this study, the phytochemical composition and biological activities of different solvent extracts (chloroform, ethanol and water) from the aerial parts of Hyoscyamus aureus L. (Solanaceae) were investigated. The analyses included the total content of phenolics and flavonoids, the antioxidant potential by the radical scavenging methods 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2,2'-azino-bis-(3-ethylbenzotrazoline-6-sulfonate) (ABTS) as well as the copper(II) ion reducing antioxidant capacity (CUPRAC), antimicrobial and antibiofilm activities by the minimum inhibitory concentration (MIC) method (against Gram-positive, Gram-negative bacteria and yeast strains) and enzyme inhibition tests for acetylcholinesterase, butyrylcholinesterase, elastase and hyaluronidase. The phytochemical profile was established using liquid chromatography-mass spectrometry/mass spectrometry (LC-MS/MS). The highest yield was obtained with HAWE (Hyoscyamus aureus water extract; 16.14 %) and the lowest with HAEE (Hyoscyamus aureus ethanol extract; 5.34 %). Among the extracts tested, HAEE had the highest total content of phenolics (25.48 μg PEs/mg extract) and flavonoids (26.66 μg QEs/mg extract) and the strongest antioxidant activity (IC₅₀ for DPPH: 46.12 µg/mL; IC₅₀ for ABTS: 33.88 µg/mL; A₀.₅: 44.26 µg/mL). In the antimicrobial experiments, the chloroform extract (HACE) showed the strongest inhibitory effect, while HAEE showed the most effective biofilm activity against Escherichia coli ATCC 25922 and Staphylococcus aureus ATCC 25923 with 11.8 % and 20.2 % inhibition, respectively. While no extract showed anti-elastase activity, HACE exhibited anti-hyaluronidase activity comparable to the standard. LC-MS/MS analysis revealed chlorogenic acid (HACE: 683.9282 ng/mL, HAEE: 15859.1327 ng/mL, HAWE: 3267.1777 ng/mL) and hesperidin (HAEE: 3974.9121 ng/mL) as major constituents of the extracts (HACE: 28.4126 ng/mL, HAEE: 7796.0074 ng/mL and HAWE: 7722.1826 ng/mL). Multivariate analyses such as heat map, Pearson correlation, principal component analysis (PCA) and hierarchical component analysis (HCA) were applied to investigate the relationships between the phytochemical constituents and biological activities. This is the first comprehensive study to investigate the antioxidant, antimicrobial, antibiofilm and enzyme inhibitory properties of H. aureus extracts in correlation with their phytochemical profiles. These results suggest that H. aureus could be a promising natural source of bioactive compounds with potential applications in the pharmaceutical, food and cosmetics industries.