High-Performance Liquid Chromatographic Determination of Catecholamines, Serotonin and Their Metabolites with Three-Potential Electrochemical Detection

Abstract

A new method of high-performance liquid chromatographic three-potential electrochemical detection was developed and applied for determining DA, DOPA, NE, E, 3MT, DOPAC, HVA, MHPG, DEP, 5HT and 5HIAA in rat-brain tissue. The proposed method employed a coulometric electrochemical detector coupled with a conditioning cell and a dualelectrode analytic cell. Three individual potentials were applied to the electrodes in order to oxidize-reduce the analytes. This detection system could effectively avoid interfering substance peaks, and could accurately quantify eleven components of catecholamines and serotonin as well as metabolites. The present method showed normal linearity with quantity injection of the standards; the detection limit was 0.17 -1.22 pg for all analytes of interest. In a measurement of neuroamines in rat-brain, the recovery of all target components averaged 74.5%.

Stability of Catecholamines, Indolamines and Related Metabolites in the Process of Sample Preparation with Ultrasonication and an Analysis by High-Performance Liquid Chromatography

“…1,2 In conclusion, most monoamines and related compounds may be decomposed by an ultrasonic physicochemical effect during the process of sample preparation. A better condition for an ultrasonic treatment includes two points: take thermo-control at 3˚C and use a short time course, such as less than 10 min.…”

“…[1][2][3][4][5] During the work of analyzing neuroamines in brain tissue, urine or plasma using HPLC with electrochemical detection (ECD), we prepared these biological samples and noticed the influence of ultrasonication, that is, it badly decreased the recovery of compounds of interest. Until recently, however, the influence of ultrasonication on the stability of catecholamines, indolamines and related metabolites had not been investigated.…”

Simultaneous determination of 3,4-dihydroxymandelic acid, 4-hydroxy-3-methoxymandelic acid, 3,4-dihydroxyphenylglycol, 4-hydroxy-3-methoxyphenylglycol, and their precursors, in human urine by HPLC with electrochemical detection

“…The technique of coulometric detection with three potentials has been described elsewhere [7]. Applying this technique, we applied 0.4 V and -0.3 V, respectively, to the conditioning cell and the second working electrode of the analytical cell.…”

“…Several publications have reported the determination of CA, IA, and their metabolites by high-performance liquid chromatography (HPLC) with different detection techniques [3][4][5][6], and many data are available about the levels of CA, IA, and their major metabolites in biological samples such as urine, tissue, and plasma [7][8][9]. There are, however, few reports of levels of the metabolites DOMA, VMA, DHPG, and MHPG, and no method has been used for simultaneous determination of CA, IA, and catecholaminergic metabolites including DOMA, VMA, DHPG, and MHPG in human urine.…”

Determination of Catecholamines and Indoleamines in Human Urine Based on Intramolecular Excimer-forming Derivatization and Fluorescence Detection

“…Liquid chromatographic (LC) methods with native fluorescence detection, 5 electrochemical detection (ECD), 6 and mass spectrometry (MS) 7 have been used for the simultaneous determination of amines. However, the native fluorescence of catecholamines is very weak.…”