Overview of the Role of Chromatographic Modes in Pharmaceutical Peptide Analysis

Peptides represent promising candidates for medical treatments. They can be designed for high specificity, which minimizes the adverse effects. Ensuring their purity, identity, and quality is crucial to their therapeutic efficacity. Peptides exhibit different physical and chemical properties (such as charge, pI, hydrophobic nature, size, etc.). These characteristics complicate the separation of a mixture of peptides. Various analytical techniques were used for peptide purification, peptide mapping, or peptide identification. HPLC continues to be the preferred method for analyzing peptides. Different HPLC modes were applied to separate peptides from their impurities and related substances. Among these modes, ion exchange, reversed-phase, normal-phase, and HILIC were performed to achieve peptide separation. This review will discuss chromatographic techniques and their role in peptide analysis, including analyzing peptide mixtures, creating peptide mapping, or isolating the peptide of interest from associated compounds.