Biogenic Silver Nanoparticles Mediated by Phytoconstituents of the Edible Spice Pimpinella anisum : Characterization and In Vitro, In Vivo, and In Silico Biological Evaluation

Abstract

ABSTRACT This study evaluated the aqueous extract of Pimpinella anisum seeds (AE‐PAS) and its green‐synthesized silver nanoparticles (PA‐AgNPs) using integrated in vitro, in vivo, and in silico approaches. LC–MS analysis identified 26 phytoconstituents, mainly sinapic acid (15.78%), hispidulin (15.35%), caffeic acid (11.09%), and rutin (8.86%). Total phenolic and flavonoid contents were 45.2 ± 2.1 mg GAE/g DE and 28.7 ± 1.5 mg QE/g DE, respectively. UV–Vis spectroscopy, scanning electron microscopy (SEM), Fourier‐transform infrared (FTIR) spectroscopy, and X‐ray diffraction (XRD) supported nanoparticle formation, indicating dry‐state particle or agglomerate features of approximately 60–90 nm, a surface plasmon resonance (SPR) peak at 421 nm, 50.81% crystallinity, and a crystallite‐domain size of 22.57 ± 1.2 nm. AE‐PAS showed measurable antioxidant and albumin‐denaturation‐inhibitory responses (DPPH IC 50 = 130.63 ± 1.3 µg/mL; albumin‐denaturation IC 50 = 721.79 ± 1.7 µg/mL). PA‐AgNPs produced measurable inhibition zones in agar well diffusion screening, but antibacterial potency could not be established because MIC and MBC were not determined. The PA‐AgNP DPPH result lacked a nanoparticle‐only blank and should be regarded as an apparent assay‐specific estimate. Hemolysis was concentration dependent, increasing from 5.20% at 50 µg/mL to 29.93% at 100 µg/mL. In behavioral models, PA‐AgNPs (0.1 mg/kg) produced measurable antidepressant‐like and analgesic responses; these findings represent preliminary behavioral evidence rather than confirmed efficacy. Molecular docking predicted favorable binding scores (−8.0 to −9.6 kcal/mol) for selected extract phytochemicals against MAO‐A/B, COX‐1/2, and TNF‐α but did not establish target inhibition or a biological mechanism. Overall, the findings are preliminary and require comprehensive physicochemical characterization, appropriate silver‐ion and material controls, quantitative antimicrobial testing, pharmacokinetic and mechanistic validation, and expanded safety assessment before biomedical translation.