3.3. Peptide Purification by Preparative Reversed-Phase High-Performance Liquid Chromatography (RP-HPLC)

Crude peptide purification was performed by RP-HPLC using a preparative SunFire C₁₈ OBD, 5 μm, 19 × 250 mm column (Waters, Milford, MA, USA at room temperature. The HPLC system (Waters) was equipped with a 2702 autosampler, a 2998 photodiode array (PDA) detector and an automated fraction collector. The lipopeptides were purified using a linear gradient of eluent A (water with 0.1% TFA) and eluent B (acetonitrile with 0.1% TFA), ranging from 20–60% B, over 25 min, at a flow rate of 10 mL/min. Purified fractions were collected and freeze-dried prior to further characterization.