To begin, prepare chitosin solution by dissolving 0.6 grams of chitosin flakes in 50 milliliters of 1%acetic acid. Agitate overnight at room temperature to get a homogenous emulsion. Add 0.5 grams of Tween 80 and stir at 60 degrees Celsius for two hours at 1, 000 RPM to get a homogenous solution.
After bringing the solution to room temperature, add 0.75 grams of bioactive compound carvacrol or thymol dropwise or gradually while stirring the mixture for 20 minutes. Add 50 milliliters of pentasodium tripolyphosphate dropwise to the mixture while stirring at room temperature. Continue stirring for 30 minutes to get a homogenous emulsion.
For purification, centrifuge the emulsion at 10, 000 G for 30 minutes at four degrees Celsius. Decant the supernatant and collect the formed nanoparticles. Wash the particles using aqueous Tween 80 with double the volume of pellets formed to remove the unbound or free bioactive compounds.
Then wash the particles with deionized water twice to get rid of impurities. Reconstitute the nanoparticles by resuspending the pellet in 30 milliliters of deionized water.
