<p>Environmental contamination by microplastics (MPs) is well documented, with potential for large-scale contamination. Public concern over the environmental impacts of MPs has led to a drastic increase in research on MP analysis; however, it is still necessary to expand existing datasets and to improve the accuracy and reliability of analytical methods. Here, we investigated a set of potential sequential step extraction methods, aiming to identify the most effective combination of density separation chemicals for maximizing MP recovery efficiency. Among these, a sequential method using water followed by sodium polytungstate was selected based on its higher performance in spiked samples, where it demonstrated higher recovery rates than a conventional single-step separation using only the densest solution. The optimized sequential method was then applied to environmental soil samples collected from a hazardous waste collection center in Gainesville, Florida. Results indicated that a greater mass of MPs was recovered using the sequential method, especially small-sized and high-density MPs that are often underestimated. This study demonstrated a sequential density separation approach as a potential effective alternative to traditional single-step methods.</p>