Sensitive high-performance liquid chromatographic method for the determination of 5-S-cysteinyldopamine, 5-S-cysteinyl-3,4-dihydroxyphenylacetic acid and 5-S-cysteinyl-3,4-dihydroxyphenylalanine.
A new HPLC method for the determination of 5-S-cysteinylcatechols has been developed. The alumina adsorbed fraction of the supernatant of brain homogenate was injected onto a reversed-phase column and a citrate-phosphate buffer containing 1-nonyl sulphate was used as mobile phase (pH 2.1). Two dual-series working electrodes of a thin-layer cell were operating together, joined by a special coupler. The assay allows determination of the 5-S-cysteinylcatechols in the striatum, limbic system and mesencephalon of one guinea pig. Recoveries of the three 5-S-cysteinylcatechols were 59-76%, whereas the limit of quantitation was 0.04-0.10 pmol. The coefficient of variation was less than 0.76-1.10% and linearity was found up to a concentration of 500 pmol. By adding ascorbic acid to the samples, artifacts resulting in HPLC peaks were either reduced in size or deleted.
Fornstedt-Wallin
B
B
Department of Pharmacology, University of Göteborg, Sweden.
Bergh
I
I
D000818:Animals; D001205:Ascorbic Acid; D001923:Brain Chemistry; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003342:Corpus Striatum / Q000737:chemistry; D003548:Cysteinyldopa / Q000031:analogs & derivatives / Q000032:analysis; D004295:Dihydroxyphenylalanine / Q000031:analogs & derivatives / Q000032:analysis; D004229:Dithiothreitol / Q000494:pharmacology; D004298:Dopamine / Q000031:analogs & derivatives* / Q000032:analysis / Q000494:pharmacology; D005260:Female; D006168:Guinea Pigs; D006863:Hydrogen-Ion Concentration; D008032:Limbic System / Q000737:chemistry; D008636:Mesencephalon / Q000737:chemistry
C092922:5-S-cysteinyl-3,4-dihydroxyphenylalanine; C049277:5-S-cysteinyl-3,4-dihydroxyphenylacetic acid; D003548:Cysteinyldopa; D004295:Dihydroxyphenylalanine; C047408:5-S-cysteinyldopamine; D001205:Ascorbic Acid; D004229:Dithiothreitol; D004298:Dopamine