Introduction of fluorescence and chemiluminescence detection to capillary chromatography based on tube radial distribution of water–hydrophilic–hydrophobic organic mixture carrier solvents

Abstract

Fluorescence and chemiluminescence detection were introduced into a tube radial distribution chromatography (TRDC) system using an open fused-silica capillary tube and a water–acetonitrile–ethyl acetate mixture carrier solution. Model analyte mixture solutions, such as Eosin Y and perylene as well as dansyl methionine and perylene, were injected into the capillary tube by a gravity method. The analyte solution was subsequently delivered through the capillary tube with the carrier solution by a microsyringe pump; the system worked under laminar flow conditions. The analytes were separated through the tube and detected by on-capillary with fluorescence detection or by end-capillary with chemiluminescence detection taking advantage of peroxyoxalate chemiluminescence reaction. Eosin Y and perylene as well as dansyl methionine and perylene were detected in this order with a carrier solution of water–acetonitrile–ethyl acetate (15 : 3 : 2 volume ratio), while they were detected in the reverse order with a carrier solution of water–acetonitrile–ethyl acetate (3 : 8 : 4 or 2 : 7 : 4 volume ratio) with fluorescence or chemiluminescence detection. The elution times of the analytes were reversed by changing the component ratio of the solvents in the carrier solution. A fluorescein isothiocyanate-labeled bovine serum albumin was also analyzed and separated from the coexisting labeling reagent with the present system.