Determination of the catecholamines and serotonin, their precursors tyrosine and tryptophan, and their main metabolites in rat brain using reversed-phase high-performance liquid chromatography with fluorimetric and oxidative amperometric detection in series.
A high-performance liquid chromatographic method for the determination of catecholamines and serotonin, their precursors and their main metabolites was developed applied to rat cerebellum, hypothalamus, striatum and cortex. A fluorimetric and an oxidative amperometric detector were used in series. For both detectors, detection limits (25-520 pg) were useful for this application, linearity of standards was excellent (average r greater than 0.9997), between-run precision for sample analytes was generally acceptable (coefficient of variation less than 10% with appreciable concentrations present) and average recoveries of standard additions to sample analytes were better than 90%. Particular attention was paid to peak identification, including both a thorough treatment of retention time agreement of peaks in standards and sample analytes, and a comparison of results for the seven compounds amenable to quantitation by both detectors. Considerable attention was also given to determining the stability of standards and sample analytes under a wide variety of conditions, and practical recommendations were made.
Chin
J R
JR
Department of Psychology, University of Newcastle, N.S.W., Australia.
D000818:Animals; D001923:Brain Chemistry; D002395:Catecholamines / Q000032:analysis*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D008297:Male; D051381:Rats; D017208:Rats, Wistar; D012015:Reference Standards; D012701:Serotonin / Q000032:analysis*; D012996:Solutions; D013696:Temperature; D014021:Tissue Preservation; D014364:Tryptophan / Q000032:analysis*; D014443:Tyrosine / Q000032:analysis*
D002395:Catecholamines; D012996:Solutions; D012701:Serotonin; D014443:Tyrosine; D014364:Tryptophan