〔Abstract〕 Objective To investigate the extraction,purification,and identification methods of Polygonum cuspidatum(P.cuspidatum)-derived exosomes and to obtain high-purity exosomes,and thus providing a reliable material foundation for their functional studies.Methods Exosomes were isolated from fresh P.cuspidatum using ultracentrifugation combined with sucrose density gradient centrifugation.Morphological characteristics,size distribution,and surface charge properties were analyzed by transmission electron microscopy(TEM),nanoparticle tracking analysis(NTA),and Zeta potential measurements.Results The purified P.cuspidatum exosomes exhibited characteristic cup-shaped or spherical bilayer morphology with a predominant size distribution of 100-200 nm,consistent with established plant exosome characteristics.The surface of these exosomes exhibits a negative charge,with the Zeta potential absolute value of root-derived exosomes being slightly higher than that of stem/leaf-derived counterparts.Conclusion The study successfully isolates and characterizes P.cuspidatum exosomes,with comprehensive analysis confirming their distinctive structural and biophysical properties.It lays a good material foundation for further exploration of its active ingredients and biological functions.