Selective trace enrichment by immunoaffinity capillary electrochromatography on-line with capillary zone electrophoresis - laser-induced fluorescence

Limited by the lack of a sensitive, universal detector, many capillary-base d liquid-phase separation techniques might benefit from techniques that ove rcome modest concentration sensitivity by preconcentrating large injection volumes. The work presented employs selective solid-phase extraction by imm unoaffinity capillary electrochromatography (IACEC) to enhance detection li mits. A model analyte, fluorescein isothiocyanate (FITC) biotin, is electro kinetically applied to a capillary column packed with an immobilized anti-b iotin-IgG support. After selective extraction by the immunoaffinity capilla ry, the bound analyte is eluted, migrates by capillary zone electrophoresis (CZE), and is detected by laser-induced fluorescence. The column is regene rated and reused many times. We evaluate the performance of IACEC for selec tive trace enrichment of analytes prior to CZE. The calibration curve for F ITC-biotin bound versus application time is linear from 10 to 300 seconds. Recovery of FITC-biotin spiked into a diluted urinary metabolites solution was 89.4% Versus spiked buffer, with a precision of 1.8% relative standard deviation (RSD).