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Subcritical water was used to provide propolis oil in water (O/W) nanoemulsions. To monitor and detect the main bioactive compounds of the prepared propolis extract, gas chromatography demonstrated that there were 47 bioactive materials in the propolis extract, among which pinostrobin chalcone and pinocembrin were the two key components. Effectiveness of two processing parameters such as the amount of saponin (0.5–2.0 g) and propolis extract (0.1–0.6 g), on particle size, polydispersity index (PDI), zeta potential, and antioxidant activity of the provided nanoemulsions, was evaluated. Results demonstrated that more desirable propolis O/W nanoemulsion, with minimum particle size (144.06 nm) and PDI (0.286), and maximum zeta potential (−21.71 mV) and antioxidant activity (90.86%) were made using 0.50 g of saponin and 0.53 g of propolis extract. Further analysis revealed that the prepared nanoemulsion based on optimum processing conditions had spherical shaped propolis nanodroplets in the colloidal solution with turbidity and maximum broad absorption peak of 0.08 a.u. and 292 nm, respectively. The prepared nanoemulsion had high antibacterial activity against both selected bacteria strains namely,  Staphylococcus aureus and Escherichia coli.

New approach in process intensification based on subcritical water, as green solvent, in propolis oil in water nanoemulsion preparation