A capillary electrophoresis (CE) method was developed to characterize protegrin IB-367, an antimicrobial peptide being developed for the treatment of oral mucositis and for other topical applications. The electrophoretic purity and levels of potential impurities/degradation products of IB-367 drug substance are determined by CE using area normalization. Electrophoresis parameters were optimized to allow optimal resolution, reproducibility and minimal analysis time. The separation and resolution between this polycationic peptide and truncated analogs determined by the CE method was much greater than those by the HPLC methods. In addition, the CE methods separates the potential impurities/degradation products from each other while the HPLC methods failed to resolve them. The CE method was validated in the aspects of accuracy, precision, linearity, range, limit of detection, limit of quantitation, specificity, system suitability and robustness. An internal standard was used for the quantitation purpose. The selection criteria of the internal standard as well as the method validation results are presented. The truncated peptide analogs were used to demonstrate the specificity of the method. These analogs were also used to evaluate the limit of quantitation of potential impurities. The relative response factors of these analogs were assessed to determine area normalization feasibility. System suitability tests were established.