[Trace analysis of biological substances by high-performance liquid chromatography with electrochemical detection]

High-performance liquid chromatography (HPLC) with electrochemical detection has been developed for the trace analysis of electrochemically active substances in biological fluids such as catecholamines. The recent development of this system, both instruments and application, has been reviewed. The principle of instrument and newly developed electrode for electrochemical detector have been described at the first section. Series electrodes, especially with coulometric detection, gave much higher selectivity to the analysis of biological substances. The application of HPLC with electrochemical detection to the determination of estrogens in biological fluids clarified the metabolic pathway of catechol estrogens. The applicability to the determination of vitamin D3 and related compounds has been also suggested. For the purpose of extending the applicability of HPLC with electrochemical detection to electro-inactive compounds, pre- and post-column labeling methods have been developed. Pre-column derivatization reagents possessing ferrocene as an electrophore provided a selective and sensitive response to the detector, because of their high-reversibility and low applied potential in the redox reaction. Post-column reaction by using immobilized enzyme reactor has been widely used for the determination of acetylcholine and choline in biological fluids.