IQUIFIB   02644
INSTITUTO DE QUIMICA Y FISICOQUIMICA BIOLOGICAS "PROF. ALEJANDRO C. PALADINI"
Unidad Ejecutora - UE
congresos y reuniones científicas
Título:
Polymeric nanoparticles as pseudostationary phase for the separation of synthetic peptides by capillary electrochromatography
Autor/es:
GRELA, DENISE A.; COLLAZO, DAIANA; VIZIOLI N
Lugar:
Lima
Reunión:
Simposio; 19th Latin-American Symposium on Biotechnology, Biomedical, Biopharmaceutical and Industrial Applications of Capillary Electrophoresis and Microchip Technology; 2013
Resumen:
Capillary electrochromatography (CEC) is habitually performed using packed, monolithic or open-tubular columns. However, the use of pseudostationary phases has turned an interesting alternative with the advantage of no requiring neither frits nor packing. The pseudostationary phase provides sites for analyte interaction and moves with or against the mobile phase. In this system, the pseudostationary phase is continuously replaced and a renewed column is used for each analysis, which avoids the contamination associated with complex matrix samples and minimizes the need of column change. [1] In this work, a technique that uses polymer nanoparticles as pseudostationary phase in CEC with UV absorbance detection is presented. The material was applied to the separation of synthetic peptides. A continuous full filling technique in which the nanoparticles were suspended in the entire electrolyte volume as well as a conventional partial filling technique was used. Nanoparticles were prepared from methacrylic acid and ethylenglycol dimethacrylate with benzoyl peroxide by utilizing a precipitation polymerization technique. The reaction was monitored by infrared spectroscopy. Polymer characterization was performed by dynamic light scattering and transmission electron microscopy. A mixture of five synthetic peptides was successfully separated using a background electrolyte constituted by acetonitrile: 25 mM phosphoric acid solution adjusted with triethanol amine to pH 3.3, 9:1, v/v. The repeatability for migration time was under 3% RSD (n= 5). The application of polymeric nanoparticles as pseudostationary phase showed to be a useful tool to improve the separation efficiency of peptides in CEC. Additional experiments are in progress to explore the technique performance in the analysis of complex protein samples.