Encapsulator

The CBD and DCA formulations were encapsulated immediately after emulsification using the gelation technique with the vibrating Encapsulator B-390 (BUCHI Labortechnik, Switzerland). The encapsulation protocol was adapted from previous studies in our group [11 (link)], with minor adjustment of the capsulation conditions: frequency range of 2000 Hz and air pressure at 950 mbar through a 200 μm nozzle with a flow regulating valve set at 2 rotations from the tightest starting point.
Prepared formulations were projected into the 100 mM CaCl₂ hardening bath, which stirred with a mild vortex, at a flow rate of 5 mL/minute and formed spherical microcapsule beads. After 10 minutes in CaCl₂, microcapsules were sieved, rinsed with deionised water and dried with a paper towel patted under the strainer. They were placed on a petri dish, covered and dried completely at 37°C for 2.5 days. Microcapsules were analysed and used for experimentation within 48 hours of drying.

Nanoemulsions with 30% w/w of cinnamon in the oil mixture and 2.5% w/w of oil mixture were prepared using the PIT method. The final composition of nanoemulsions was set to 2.5% w/w of oil mixture, 10% w/w of surfactant mixture and 87.5% w/w of chitosan aqueous solution at 1% w/w as the continuous phase. Due to the high viscosity of the chitosan solution, the Buchi Encapsulator could not be used. Therefore, the beads were prepared by dropping the chitosan solution into the 5% w/w TPP solution with a hand pipette. All the chitosan beads were left in the TPP solution during 10 min to assure a proper gelation. Some of the beads were reserved as single Ch beads, and the rest of the beads continued the second coating process. These beads were incubated in an alginate 1% w/w solution for 10 more minutes. Some of the resulting beads were set apart and named Ch-Alg beads, and the rest were submerged into a new 1% w/v CaCl₂ bath for 10 min in order to gel the shell alginate coating (Ch-Alg-CaCl₂ beads).