Display omitted •We developed experimental method for quantifying riverine macroplastic fragmentation.•Mass loss of a riverine macroplastic item used is as a proxy for its fragmentation rate.•This method give an opportunity to track macroplastic fragmentation across various rivers.•Our method can be adapted to study macroplastic fragmentation in other environments. Direct field measurements of macroplastic fragmentation during its transport in rivers are currently unavailable, and there is no established method to perform them. Previous studies have showed that macroplastic fragmentation results in the production of harmful microplastics, and river channels can be hotspots for this process. Therefore, obtaining information about this process is crucial for quantifying the production of secondary microplastics in rivers and assessing the related risks for riverine biota and human health. Here, we propose a simple low-cost methodology for quantifying riverine macroplastic fragmentation by conducting repeated measurements of the mass of tagged macroplastic items before and after their transport in the river. As a proof-of-concept for this method, we conducted a 52–65 day experiment that allowed us to measure a median fragmentation rate of 0.044 ± 0.012 g for 1-liter PET bottles during their transport at low to medium flow in the middle mountain Skawa River in the Polish Carpathians. Using the obtained data (n = 42), we extrapolated that during low to medium flows, the median yearly mass loss of PET bottles in the study section is 0.26 ± 0.012 g/year (0.78 ± 0.036 % of bottle mass), and the median rate of bottle surface degradation is 3.13 ± 0.14 μm/year. These estimates suggest a relatively high fragmentation rate for a PET bottle in a mountain river even under low to medium flow conditions without high-energy transport. We discuss how our simple and relatively low-cost methodology can be flexibly adapted and future optimized to quantify macroplastic fragmentation in various types of rivers and their compartments, informing future mitigation efforts about the rates of formation and dispersion of secondary microplastics.