Aim: The aim of this research study was to develop a simple, rapid, precise, accurate and economical RP-HPLC method, with a simple mobile phase for the identification and determination of the antimicrobial peptide α-defensin-2. Materials and Methods: Separation was carried out at 25°C, using column Luna 5U (C18, 250x4.6, 5 μm) with mobile phase consisting of acetonitrile: water (40: 60 v/v). The detector was set at 210 nm. The flow rate was 1.2 ml/min and injection volume was 20 μl. Results: The developed method was validated as per ICH guidelines and the kinetic behavior of α-defensin 2 was studied in respect of different pH. Conclusion: The obtained data can serve for creating an in vivo monitoring program and quality control for different studies and preparations.
Key words: Antimicrobial peptides, α-defensin 2, HPLC, Chemical stability, Analysis.