Determination of peptides by capillary electrophoresis-electrochemical detection using on-column Cu(II) complexation.

A Cu(II)-coated capillary has been developed for the determination of peptides by capillary electrophoresis with electrochemical detection. Capillaries were prepared by forcing a solution 48 microM in CuSO4, 120 microM in tartaric acid, 2.4 mM in NaOH and 120 microM in KI through them for 25 min; the resulting capillaries are stable for at least 12 h. Under alkaline conditions, peptides complex with Cu(II) present on the walls of the capillary to form Cu(II)-peptide complexes which can be detected oxidatively at a carbon fiber electrode. Di-, tri-, tetra- and pentaglycine were determined with a detection limit of 7 x 10(-7) M for triglycine. N-Terminal-blocked peptides can also be determined via this method. This system is more sensitive than direct detection of peptides by UV at 210 nm and exhibits higher selectivity than commonly employed derivatization procedures based on reactions with a primary amine functionality.