Liquid chromatography with amperometric reaction detection involving electrogenerated reagents: applications with in-situ generated bromine.
We describe the use of electrogenerated reactants for continuous on-line reaction detection with thin-layer hydrodynamic amperometry. The reagent is introduced into the liquid-chromatographic column effluent at a constant rate by using controlled-current electrochemistry. After the effluent passes through a short reaction coil, the reagent concentration is monitored at the detector. Reaction of eluted compounds with bromine is signalled by changes in the current detected. The direct electrochemical control of the reagent concentration allows changes to be made, even during the course of obtaining a chromatogram. Depending on the specific reagent or reaction, the reagent is supplied either by addition of a second stream or by direct generation in the mobile phase. The latter configuration provides sufficient baseline stability to permit detection of the uptake of as little as 10 pmol of reagent bromine. The technique has been used to detect nanograms of underivatized fatty acids, prostaglandins, and phenols after separation by reversed-phase liquid chromatography.
D001966:Bromine; D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation / Q000379:methods*; D005227:Fatty Acids / Q000032:analysis*; D007202:Indicators and Reagents; D010636:Phenols / Q000032:analysis*; D011453:Prostaglandins / Q000032:analysis*; D013438:Sulfhydryl Compounds / Q000032:analysis; D014815:Vitamins / Q000032:analysis*
D005227:Fatty Acids; D007202:Indicators and Reagents; D010636:Phenols; D011453:Prostaglandins; D013438:Sulfhydryl Compounds; D014815:Vitamins; D001966:Bromine