On-line sample stacking of peptides in capillary electrophoresis for the study of prebiotic reactions between alpha,alpha-dialkylated amino acids and amino acid N-carboxyanhydrides.
The reaction between alpha,alpha-dialkylated amino acids and amino acid N-carboxyanhydrides is slow leading to low concentrations of products (peptides). The detection by capillary electrophoresis of the analytes contained in such samples is therefore a challenging issue. In this work, on-line sample pre-concentration methods based on field-amplified sample stacking have been implemented and compared. Because of the high ionic strength present in the sample matrix, samples were diluted with an organic solvent prior to analysis to decrease the sample conductivity. Different modes of sample injection (field amplified sample injection (FASI), hydrodynamic normal sample stacking (NSS) or large volume sample stacking (LVSS)) were compared. Pre-concentration factors of 20 for FASI, about 30-40 for NSS and 60 for LVSS were obtained for the analysis of (l,l) dipeptide of valine in a large excess of isovaline and 0.2M of ionic strength. For LVSS application and resolution optimisation, a new non-covalent coating based on the partial modification of the capillary surface was used to tune the electroosmotic flow magnitude and to pump the sample matrix out of the capillary. This on-line sample pre-concentration step allowed confirming that oligopeptides including alpha,alpha-dialkylated amino acids are formed during the reaction between alpha,alpha-dialkylated amino acids and N-carboxyanhydride amino acids.
Danger
Grégoire
G
Institut des Biomolécules Max Mousseron, UMR 5247 CNRS - Université de Montpellier 1 - Université de Montpellier 2, Place Eugène Bataillon CC 1706, 34095 Montpellier Cedex 5, France. dangergregoire@yahoo.fr
Pascal
Robert
R
Cottet
Hervé
H
D000596:Amino Acids / Q000737:chemistry*; D053841:Electroosmosis; D019075:Electrophoresis, Capillary / Q000379:methods*; D009862:Online Systems; D010455:Peptides / Q000737:chemistry*
D000596:Amino Acids; D010455:Peptides