Eco-synthesis, process optimisation, and therapeutic assessment of cow urine distillate derived copper nanoparticles

The present research successfully validates a sustainable, straightforward, cost-effective, and non-toxic biosynthesis of biologically active copper nanoparticles (CuNPs) using cow urine distillate (CUD), a traditional component of Vedic medicine. The study also investigates the effect of different process variables: reductant concentration, pH, reaction temperature, and reaction time, employing a factorial design approach (Box-Behnken design) on the synthesis of CUD-CuNPs. UV-Vis spectroscopy confirmed nanoparticle formation, showing surface plasmon resonance peaks between 269 and 275 nm. Morphological analysis using Transmission Electron Microscopy and Scanning Electron Microscopy-Energy Dispersive X-ray demonstrated the formation of spherical nanoparticles. XRD analysis showed a crystalline structure with prominent reflections at 43.4°, 50.5°, and 74.2°. The synthesized CUD-CuNPs had an average size of 232.76 nm, a polydispersity index of 0.421, and a zeta potential of -16.45 mV. Antimicrobial assessment of CUD-CuNPs using the agar well diffusion assay demonstrated their bactericidal potential against Staphylococcus epidermidis and Pseudomonas aeruginosa, with remarkable inhibitory zones of 25.66 ± 0.57 mm. Significant inhibition was also observed against E. coli (25.66 ± 0.57 mm) and B. subtilis (25.25 ± 1.00 mm). Furthermore, CUD-CuNPs significantly suppressed the activity of α-amylase and α-glucosidase enzymes, with IC₅₀ values of 30.88 ± 1.07 µg/mL and 24.94 ± 1.02 µg/mL, respectively. In DPPH, nitric oxide, and H₂O₂ scavenging assays, CUD-CuNPs exhibited considerably stronger radical scavenging activity than CUD. This enhanced potency indicates a promising avenue for utilizing biosynthesized CUD-CuNPs in developing therapeutic drugs.