Precolumn Reactions with Electrophoretic Analysis Integrated on a Microchip

Abstract

A glass microchip was constructed to perform chemical reactions and capillary electrophoresis sequentially. The channel manifold on the glass substrate was fabricated using standard photolithographic, etching, and deposition techniques. The microchip has a reaction chamber with a 1 nL reaction volume and a separation column with a 15.4 mm separation length. Electrical control of the buffer, analyte, and reagent streams made possible the precise manipulation of the fluids within the channel manifold. The microchip was operated under a continuous reaction mode with gated injections to introduce the reaction product onto the separation column with high reproducibility (<1.8% rsd in peak area). The reaction and separation performances were evaluated by reacting amino acids with o-phthaldialdehyde to generate a fluorescent product which was detected by laser-induced fluorescence. Control of the reaction and separation conditions was sufficient to measure reaction kinetics and variation of detection limits with reaction time. Half-times of reaction of 5.1 and 6.2 s and detection limits of 0.55 and 0.83 fmol were measured for arginine and glycine, respectively. 18 refs., 10 figs.