Ultrasonic-Assisted Extraction of Bee Pollen as Antioxidant Activity for Develop to Nanoemulsion in Topical Formulation by Box-Behnken Design
DOI:
https://doi.org/10.48048/tis.2026.13729Keywords:
Bee pollen extract, Antioxidant activity, Cytotoxicity, Nanoemulsion, Box-Behnken design, Response Surface MethodologyAbstract
Bee pollen is a natural supplement composed of flower pollen, nectar and the secretions of bees. This research analyzed the composition of bee pollen using FTIR-ATR, which showed that bee pollen contains components that enhance antioxidant activity such as protein. The antioxidant potential of bee pollen extract was evaluated. Analyses on bee-pollen from ultrasonic extraction exhibited impressive radical scavenging properties in DPPH, ABTS and NO assay, without being a cell-toxic, and own potential for the investigations into gene expression and inflammatory cytokine modulation. For developing nanoemulsion in formulation, Box-Behnken Design using Response Surface Methodology (RSM) was used to determine the significant stability variables of particle size (PS), viscosity, and type of surfactant Tween. Phase separation was observed in 8 of 17 experimental trials, where higher pressure (7,000 psi) increased particle size and higher surfactant concentrations slightly enlarged particles but reduced zeta potential. Time exhibited a negligible impact. A combination of 2% surfactant and 5,000 psi pressure applied for 30 min was the optimal process condition, producing nanoparticles of 140.46 nm with a corresponding ZP of −24.46 mV, as corroborated by triplicate measurements. Formulations 1 - 3, 6, 9, 12 - 15, and 17 passed all stability tests. Conversely, formulations 4 - 5, 7 - 8, 10 - 11, and 16 showed phase separation and need further modification. These results are in accordance with known stiffness stabilization mechanisms in nanoemulsion systems, evidencing a surfactant assisted particle dispersion enhancement. This 2-dimensional strategy further promotes the functional possibility of bee pollen through powerful antioxidant profiling and rationally designed delivery systems.
HIGHLIGHTS
- Antioxidant Properties and Safety: The ultrafiltration-extracted bee pollen expressed significant antioxidant capacity in DPPH, ABTS, and NO scavenging assays as well as no cytotoxicity, providing scope for investigating a cellular level for gene expression and inflammatory cytokines.
- Factors influencing stability of nanoemulsions: Box-Behnken design (BBD) optimization identified that the particle size, viscosity, and surfactant type (i.e., Tween) were associated with the phase separation among 8 of the 17 nanoemulsions formulated. The higher surfactant concentration slightly increased the particle size and decreased the zeta potential, and pressure (3,000 - 7,000 psi) changes inversely affected particle size.
- Optimal Formulation Parameters: RSM study resulted in surfactant level of 2%, pressure of 5,000 psi and processing time of 30 min for enhanced predicted and validated particle size (140.46 nm) and zeta potential (−24.46 mV) The combined optimized formulation parameters were validated experimentally with least deviation.
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