HPLC Analysis of Extracellular Vesicles

HPLC analysis of the EV is performed following [57 (link)] with some modifications. The SCRE was analyzed using an Agilent 1260 series instrument and Eclipse C18 column (4.6 mm × 250 mm i.d., 5 µm). Separation was performed at a flow rate of 0.9 mL/min. The mobile phase consisted of water (reservoir A) and 0.5% trifluoroacetic acid in acetonitrile (reservoir B) at a concentration of 0.1%. The mobile phase was sequentially programmed with a linear gradient as follows: 0 min (82%A); 0–5 min (80%A); 5–8 min (60%A); 8–12 min (60%A); 12–15 min (82%A); 15–16 min (82%A); and 16–20 (82%A). A multi-wavelength UV detector was used for detection at 280 nm. The injection volume for each sample solution was 5 μL. The column temperature was retained at 40 °C.