Preparation of KO Emulsions
The creation of KO emulsions followed a two-phase procedure. In the aqueous phase, a 2.5% wt% stabilizer consisting of 65% sodium caseinate, 6% gum arabic, and 29% Tween-20 was added to a pH 7 phosphate buffer solution. This mixture was stirred for 2 h to ensure complete dissolution. Simultaneously, the oil phase was formed by dispersing 0.3g of KO in a 20 mL solution comprising dichloromethane and acetone in a 1:2 v/v ratio. The two phases were combined at a 1:9 ratio and stirred at 40 ℃ for 1 hour, resulting in the formation of a coarse emulsion. This crude emulsion was further sheared at speeds ranging from 800 to 1000 r/min for 5 to 7 minutes. Subsequently, a high-pressure homogenization (HPH) process was implemented, involving three rounds of homogenization at 800 bar (equivalent to 80 MPa). To remove any residual organic solvent, vacuum processing was employed (Kaya et al., 2021).
In parallel, a separate set of KO emulsions was produced using a (Langfang General Machinery Manufacturing Co. Ltd., Langfang, China) microfluidizer by following established protocols detailed in prior research. Essentially, this involved homogenizing a 10% w/w oil phase with a 90% w/w water phase. The oil and water phases were individually introduced into the dual-channel microfluidized bed through glass reservoirs. Upon convergence, they underwent high-pressure conditions of 80 MPa and were subjected to three cycles of this process (Zhao et al., 2021).