Optimization of Setpoint Conditions for Enhanced Biofabrication of Silver Nanoparticles Using Helichrysum crispum...

This study investigated the optimization of setpoint conditions used for the enhanced biofabrication of silver nanoparticles (H.C-AgNPs) using Helichrysum crispum extracts. A Box-Behnken Design (BBD) model was used to evaluate the effects of reaction time, temperature, an H. crispum extraction volume, and a 0.1 M AgNO₃ solution volume. A second-order polynomial regression equation was developed with a high R² of 0.9629, indicating that the model explained 96.29% of the variability in the data. The statistical significance of the model was confirmed with an F-value of 25.92 and a p-value of less than 0.0001. The optimal biofabrication conditions were determined to be a reaction time of 60 min, a temperature of 50 °C, an H. crispum extract volume of 10 mL, and a silver nitrate volume of 90 mL, achieving a peak absorbance of 3.007 a.u. The optimized conditions were experimentally validated, resulting in an absorbance of 3.386 a.u., reflecting a 12.6% increase. UV-Vis spectroscopy showed a distinct surface plasmon resonance (SPR) peak at 433 nm. XRD analysis confirmed a crystalline face-centered cubic (FCC) structure with a primary diffraction peak at 2θ = 38.44° (111 plane). SEM and EDS results confirmed a uniform size and high purity, while FTIR spectra confirmed the involvement of phytochemicals in nanoparticle stabilization. TEM analysis revealed a uniform particle size distribution with a mean size of 19.46 nm and a dispersity of 0.16%, respectively. These results demonstrate the importance of statistical tools in optimizing the setpoint conditions used in the biofabrication of AgNPs, which have applications in various fields.