Simultaneous Determination of 3,4‐Dihydroxyphenylalanine, 5‐Hydroxytryptophan, Dopamine, 4‐Hydroxy‐3‐Methoxyphenylalanine, Norepinephrine, 3,4‐Dihydroxyphenylacetic Acid, Homovanillic Acid, Serotonin, and 5‐Hydroxyindoleacetic Acid in Rat Cerebrospinal Fluid and Brain by High‐Performance Liquid Chromatography with Electrochemical Detection

Abstract

A method using reversed-phase ion-pair high-performance liquid chromatography with electrochemical detection for the simultaneous determination of tryptophan (TRP), 3,4-dihydroxyphenylalanine (DOPA), and their metabolites in whole brain, small-brain parts, and cerebrospinal fluid of rats has been developed. The sample preparation requires only homogenization in perchloric acid and centrifugation before injection onto the column. With a LiChrosorb RP-18 (10 micrometer) column and a mobile phase consisting of a phosphate (NaH2PO4, 0.1 M)-methanol mixture with octylsulfonate (2.6 x 10(-3) M) at pH 3.35 and 26 degrees C, the separation of DOPA, dopamine, norepinephrine, 3,4-dihydroxyphenylacetic acid, homovanillic acid, 4-hydroxy-3-methoxyphenylalanine, TRP, 5-hydroxytryptophan (5-HTP), serotonin, and 5-hydroxyindoleacetic acid was achieved. The method has been applied to study the effect of alpha-monofluoromethyldopa alone and in combination with L-DOPA or L-5-HTP, on the catechol and 5-OH indole levels in brain and CSF of the rat.