Simple Sheath Flow Reactor for Post-column Fluorescence Derivatization in Capillary Electrophoresis

Abstract

A system for post-column fluorescence derivatization in capillary electrophoresis is described. The post-column reactor uses a sheath flow detection cell where the reagents, o-phthaldialdehyde and β-mercaptoethanol, are added to the sheath buffer and mix by diffusion with the analytes effusing from the separation capillary. Reaction progress is monitored and optimized by imaging a large portion of the sheath flow cuvette using an extended UV source and a CCD camera. Significantly, this design provides the ability to switch between the analysis of pre- and post-column derivatized amino acids and peptides easily and without sacrificing system performance. The lack of turbulent flow in this system minimizes post-separation band broadening. The limit of detection for glycine is 9.4 × 10^–8 M (110 amol) with a separation efficiency of 190000 theoretical plates, without stacking. The performance of the system for a series of amino acids was evaluated using post-column and pre-capillary derivatization.