Determination of catecholamines by flow-injection analysis and high-performance liquid chromatography with chemiluminescence detection

Abstract

A chemiluminescence (CL) detection of catecholamines [norepinephrine (NE), epinephrine (E), dopamine (DA) and L-dopa (LD)] is described for the flow-injection (FI) and high-performance liquid chromatographic (HPLC) determination of these compounds. The detection method is based on the inhibition effect of catecholamines (CAs) on the CL reaction of luminol with iodine in the alkaline medium. The proposed FI method allows the determination of CAs in pharmaceutical preparations for the purpose of drug quality control. The calibration curves show good linearity in the concentration range of: 1.1-20.0 microg l(-1) (NE), 0.5-5.0 microg l(-1) (E), 0.6-9.0 microg l(-1) (DA) and 0.6-10.0 microg l(-1) (LD). The limits of detection (defined as a signal-to-noise ratio of 3) are: 0.34 microg l(-1) (NE), 0.15 microg l(-1) (E) and 0.18 microg l(-1) (DA, LD). The HPLC procedure was successfully applied for the determination of catecholamines (NE, E, DA) in human urine after solid-phase extraction (SPE). In a simple run time CAs can be determined in 20 min. The chromatographic linear ranges are: 5.0-72.0 microg l(-1) (NE), 5.0-48.0 microg l(-1) (E) and 5.0-96.0 microg l(-1) (DA). The limits of detection for three urinary CAs are: 0.71 microg l(-1) (NE), 0.26 microg l(-1) (E) and 0.73 microg l(-1) (DA).