Blueberry exosome‐like nanoparticles extraction guidelines: Comparative methods and standardized operations

Abstract Blueberry‐derived exosome‐like nanoparticles (BENs) hold significant potential for functional foods and biomedical applications. Efficient extraction is crucial for exploiting their benefits. This guideline compares four principal methods based on key operational and performance parameters. Ultracentrifugation (100,000 g, 70 min) yields high‐purity BENs but requires costly equipment and may impair vesicle integrity. Polymer precipitation (8% PEG6000, overnight incubation) offers high yield and cost‐effectiveness, yet purity is compromised by polymer and co‐precipitant residues. Density gradient centrifugation (sucrose gradients, 100,000 g, 16 h) achieves the highest purity and subpopulation resolution but is time‐consuming, low‐throughput, and unsuitable for scale‐up. Size exclusion chromatography provides rapid, activity‐preserving separation with good reproducibility, though sample load is limited and initial setup costs are considerable. The standardized application of these methods ensures reproducible BENs quality, facilitating their use in research and industry.