Separation of peptides by open-tubular capillary electrochromatography using Fe(III)-deuteroporphyrin as a covalently attached stationary phase

YONE A; CARBALLO R; REZZANO IN; VIZIOLI NM

Sevilla, España

Simposio; 15th Symposium on Biomedical, Biopharmaceutical and Industrial Applications of Capillary Electrophoresis and Microchip Technology; 2009

Universidad CEU-San Pablo

Proteins and peptides belong, together with nucleic acid, to the most important biologically active compounds, since they play a fundamental role in control and regulation of life process of all living organisms. In the current era of proteomics, the structure and function of many proteins are identified by means of their peptide fragments. At the same time, and due to their essential role in biological functions, peptides are a group of biopharmaceuticals with growing demand as therapeutic agents. For all these reasons, the importance of peptides is really recognized and their qualitative and quantitative analyses are greatly valued and remain one of the most challenging application field of capillary electromigration methods. This presentation shows the separation of seven biologically active peptides by open-tubular capillary electrochromatography (OT-CEC) in fused-silica (FS) capillaries chemically modified with Fe(III)-deuteroporphyrin using UV-absorption detection. The effect of background electrolyte pH and content of organic solvent modifier was investigated. Considering the method sensitivity, lower concentration limits of detection were obtained for all peptides in their OT-CEC separation as compared to their capillary zone electrophoresis separation in bare FS capillary. The Fe(III)-deuteroporphyrin column proved to be highly stable over time and showed acceptable precision of migration times and corrected peak areas (relative standard deviation in the range 2 - 4 %).