Microplastics (MPs) have already spread across the globe and have been found in drinking water and human tissues. This may pose severe threats to human health and water environment. Therefore, this study accurately evaluated the removal effect of metal-modified biochar on polystyrene microplastics (PS-MPs) (1.0 μm) in the water environment using a high-throughput fluorescence quantification method. The results indicated that Fe-modified biochar (FeBC) and Fe/Zn-modified biochar (Fe/ZnBC) had good removal efficiencies for PS-MPs under the dosage of 3 g/L, which were 96.24% and 84.77%, respectively. Although pore effects were observed (such as “stuck”, “trapped”), the electrostatic interaction was considered the main mechanism for the adsorption of PS-MPs on metal-modified biochar, whereas the formation of metal-O-PS-MPs may also contribute to the adsorption process. The removal efficiency of PS-MPs by FeBC was significantly reduced under alkaline conditions (pH = 9 and 11) or in the presence of weak acid ions (PO₄³⁻, CO₃²⁻, HCO₃⁻). A removal efficiency of 72.39% and 78.33% of PS-MPs was achieved from tap water (TW) and lake water (LW) using FeBC when the initial concentration was 20 mg/L. However, FeBC had no removal effect on PS-MPs in biogas slurry (BS) and brewing wastewater (BW) due to the direct competitive adsorption of high concentrations of chemical oxygen demand (COD). The findings of this study highlighted that metal-modified biochar had a potential application in purifying tap water or lake water which contaminated by MPs.