<p>This study aimed to compare the physicochemical characteristics and lipid composition of cabbage-derived extracellular vesicles (EVs) isolated using ultracentrifugation (UC) and ultrafiltration (UF) method. The UF process effectively removed non-vesicular impurities in a stepwise manner, eventually obtaining EVs isolated from cabbage with particle sizes, concentrations, and purities comparable to those isolated using UC. The particle size was measured to be 83.33&#xa0;nm for UC and 95.33&#xa0;nm for UF. The particle concentrations were 1.30 × 10¹¹ and 2.64 × 10¹¹ particles/mL, respectively. Both methods showed typical EV morphologies under a microscope. Additionally, protein marker analysis using a commercial mammalian exosome ELISA detected the surface tetraspanins CD9 and CD63 in EV fractions from both UC and UF. Regarding phospholipid composition, the proportion of phospholipids reached the highest levels of 90.10% in UF and 85.37% in UC. In conclusion, based on the analysis of physicochemical properties and phospholipid composition, UF can be considered an effective alternative method for isolating EVs with physicochemical characteristics comparable to those obtained using UC.</p>