4.3. Purification of Mastoparan Peptides

Twelve lyophilized venom sacs were extracted (5 × 1 mL) with 1: 1 acetonitrile-water containing 0.1% TFA (CH₃CN/H₂O/0.1% TFA), and the extracts were subjected to reverse-phase HPLC (Waters Associates, Milford, MA, USA) using CAPCELL PAK C₁₈, 10 × 250 mm (SHISEIDO Co., Ltd., Tokyo, Japan) with linear gradient from 5% to 95% CH₃CN/H₂O/0.1% TFA at a flow rate of 2.5 mL/min over 30 min. The peaks eluted at 21 and 22 min were manually collected (Figure 1B).