Health and security concerns have made it essential to develop integrated, continuous collection and sensing platforms that are compact and capable of real-time detection. In this study, we numerically investigate the flow physics associated with the single-step collection and enrichment of aerosolized polystyrene microparticles into a flowing liquid using a stratified air-water flow in a U-shaped microchannel. We validate our simulation results by comparing them to experimental data from the literature. Additionally, we fabricate an identical microfluidic device using PDMS-based soft lithography and test it to corroborate the previously published experimental data. Diversion and entrapment efficiencies are used as evaluation metrics, both of which increase with increasing particle diameter and superficial air inlet velocity. Overall, our ANSYS Fluent two-dimensional (2D) and three-dimensional (3D) multiphase flow simulations exhibit a good agreement with our experimental data and data in the literature (average deviation of ∼11%) in terms of diversion efficiency. Simulations also found the entrapment efficiency to be lower than the diversion efficiency, indicating discrepancies in the literature in terms of captured particles. The effect of the Dean force on the flow physics was also investigated using 3D simulations. We found that the effect of the Dean flow was more dominant relative to the centrifugal force on the smaller particles (e.g., 0.65 μm) compared to the larger particles (e.g., 2.1 μm). Increasing the superficial air inlet velocity also increases the effect of the centrifugal forces relative to the Dean forces. Overall, this experimentally validated multiphase model decouples and investigates the multiple and simultaneous forces on aerosolized particles flowing through a curved microchannel, which is crucial for designing more efficient capture devices. Once integrated with a microfluidic-based biosensor, this stratified flow-based microfluidic biothreat capture platform should deliver continuous sensor-ready enriched biosamples for real-time sensing.