In the current research work, the flow behavior of a liquid–liquid extraction (LLE) process in a serpentine microchannel was analyzed. The simulation was performed using a 3D model and the results were found to be consistent with experimental data. The impact of the flow of chloroform and water on the flow model was also examined. The data indicate that once the aqua and organic phases flow rates are low and similar, a slug flow pattern is observed. However, as the overall flow rate raises, the slug flow transforms into parallel plug flow or droplet flow. An increment in the aqua flows while maintaining a constant organic phase flow rate results in a transition from slug flow to either droplet flow or plug flow. Finally, the patterns of flow rate in the serpentine micro-channel were characterized and depicted. The results of this study will provide valuable insights into the behavior of two-phase flow patterns in serpentine microfluidic devices. This information can be used to optimize the design of microfluidic devices for various applications. Furthermore, the study will demonstrate the applicability of CFD simulation in investigating the behavior of fluids in microfluidic devices, which can be a cost-effective and efficient alternative to experimental studies.