This study focused on the formulation and characterization of silver nanoparticles (AgNP) functionalized with d-limonene. The nanoparticles were functionalized by phase inversion and the synthesis of the nanoparticles was performed in situ; particle size was determined by laser diffraction, zeta potential and optical colloidal stability using Multiscan 20 for a period of 24 hours at 37 °C; the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of the formulated material on Escherichia coli ATCC 25922, Staphylococcus aureus ATCC 29213, Klebsiella oxytoca ATCC 700324, Enterococcus casseliflavus ATCC 700327, Escherichia coli BLEE, carbapenem-resistant Pseudomona aeruginosa were determined. The nanoparticles showed colloidal stability at a d-limonene concentration of 3.93%, silver ions at 1.61 × 10⁻³%, non-ionic adjuvant at 24% and ascorbic acid at 5.88%; citric acid/citrate (1:1) 0.48M for a pH of 4.5 was used as a buffer system. The formulation was classified as a polydisperse system (PD = 0.0851), with a zeta potential of −11.6 mV and average particle size of 81.5 ± 0.9 nm. A particle migration velocity of −0.199 ± 0.006 mm∙h⁻¹, a constant transmission profile and backscattering profile with variations of 10% were evidenced, which represents a stable formulation. The nanoparticles presented an MIC and an MBC of 28 μg∙mL⁻¹ (5.6 × 10⁻²% d-limonene and 4.7 × 10⁻⁵% AgNP) against all tested bacteria.

This work shows the results of the biosynthesis of silver nanoparticles using the microalga Chlorella sp, using growth media with different concentrations of glycerol, between 5%–20%, and different light and temperature conditions. The synthesis of nanoparticles was studied using supernatants and pellets from autotrophic, heterotrophic and mixotrophic cultures of the microalga. The presence of nanoparticles was verified by ultraviolet-visible spectroscopy and the samples showing the highest concentration of nanoparticles were characterized by scanning electron microscopy. The mixotrophic growth conditions favored the excretion of exopolymers that enhanced the reduction of silver and thus the formation of nanoparticles. The nanoparticles obtained presented predominantly ellipsoidal shape with dimensions of 108 nm × 156 nm and 87 nm × 123 nm for the reductions carried out with the supernatants of the mixotrophic cultures with 5% and 10% glycerol, respectively.