Role of HILIC Stationary Phases in Pharmaceutical Peptide Analysis
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Abstract
Hydrophilic interaction chromatography (HILIC) is characterized by a polar stationary phase and an organic (apolar) mobile phase containing a small proportion of water. The HILIC retention mechanism is based on solute partitioning between the mobile phase (rich in organic solvents) and the aqueous phase layer. This separation method had already been used in 1975 for the analysis of oligosaccharides. The term HILIC was proposed by Alpert in 1990 during a study on the separation of amino acids and peptides. Stationary phases specifically developed for HILIC approaches can be particulate (pure silica or polar groups grafted onto silica-based or polymeric supports) or monolithic. This chromatographic mode has proven useful, particularly for hydrophilic peptides exhibiting low retention on RP columns. This review aimed to present the role of HILIC particulate and monolithic phases in the analysis of pharmaceutical peptides. Detailed descriptions of different HILIC phases were presented. Examples of peptide separation by HILIC mode were shown.
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