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.