The increasing demand for sustainable biomedical solutions has driven ongoing research toward green nanomaterial synthesis. The present study reported an eco-friendly, cost-effective protocol for synthesizing silver nanoparticles (AgNPs) using the aqueous leaf extract of the ethnomedicinal plant Bryophyllum pinnatum (Lam.). The reduction of Ag⁺ ions to metallic silver was confirmed by a distinct color transition from pale yellow to brown. Fourier Transform Infrared Spectroscopy (FTIR) analysis has been performed, validating the dual role of plant phytoconstituents. Significant absorption bands at ~3400 cm⁻¹ (O–H stretching) and ~1635 cm⁻¹ (C=O stretching) indicated that phenolic compounds, flavonoids and proteins were present within the extract. These functioned as effective bio-reductants and stabilizing capping agents for the nanoparticles. Further, antibacterial potential of the synthesized AgNPs was assessed against Gram-negative (Pseudomonas aeruginosa MTCC 2295 and Escherichia coli MTCC 5704) and Gram-positive (Staphylococcus aureus MTCC 3160, and Bacillus subtilis MTCC 121) bacteria by using agar-well diffusion method. The AgNPs was exhibited significantly greater antibacterial activity than the crude leaf extract. B. subtilis MTCC 121 has shown highest susceptibility with an inhibition zone of 21 ±1.9 mm while E. coli showed a zone of 20 ±1.7 mm. The AgNPs was exhibited slightly lower inhibition zones than standard antibiotic Ciprofloxacin (25–28 mm). However, their broad-spectrum efficacy was attributed to the synergistic interplay between the nanoparticles specific surface area and bioactive capping agents. The two-way ANOVA showed a highly significant effect of treatment type (p < 0.001) was represent that the synthesised B. pinnatum (Lam.) extract-derived AgNPs have superior antibacterial activity compared with the B. pinnatum (Lam.) extract alone. There was also a significant difference among bacterial strains (p < 0.05) indicates wide-ranging susceptibility. The prospects of the present findings endorsed B. pinnatum (Lam.) extract-derived AgNPs as promising candidates for developing natural and effective antimicrobial agents. Keywords: Silver Nanoparticles (AgNPs), Bryophyllum pinnatum (Lam.), Green Synthesis, Antibacterial Activity, Eco-friendly
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