The production of crude oil is always accompanied by water production, which may create severe separation problems. It is important to understand the stabilization mechanism and parameters contributing to the formation of emulsion, specifically the synergy mixing of surfactants. These factors have not been studied primarily in previous studies. The main objective of the current work was to assess the influence of synergy mixing of nonionic surfactants, sorbitan monooleate (hexitol) and polysorbate 80 (glycol), which are mainly affecting the stability of oil-in-water emulsions. Several factors, such as the mixing rate, mixing time, and aging time of the studied emulsions were also investigated. Response surface methodology (RSM), and central composite design (CCD) were employed to design the experiments. Emulsion stability was measured through a static bottle test over a range of time (1-7 days) at a temperature of 60 °C. A model was established with a coefficient of determination value at 0.8814 and the highest emulsion stability achieved was 42.83. The least water separation was observed at 0.5 v/v hexitol, 1.5 v/v glycol, 15 000 rpm mixing rate in 5 minutes, and seven-day ageing time to achieve �41.56 emulsion stability. The minimum emulsion stability of �25.0 was observed using 0.5 v/v of sorbitan monooleate and polysorbate 80 at 5000 rpm of mixing rate in 15 min and under seven days of observation. The results also revealed that the mixing time and ageing time do not affect the stability of the prepared emulsions. Hexitol, mixing rate, synergy mixing of nonionic surfactants and polysorbate 80, and mixing speed significantly influence emulsion stability. The R2 value of 88.14 verified that the model is well-fitted and the optimal values for the input variables were successfully obtained using RSM. © 2022 The Royal Society of Chemistry.