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.
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.
D015102:3,4-Dihydroxyphenylacetic Acid / Q000134:cerebrospinal fluid; D006916:5-Hydroxytryptophan / Q000134:cerebrospinal fluid; D000818:Animals; D001923:Brain Chemistry; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D004295:Dihydroxyphenylalanine / Q000134:cerebrospinal fluid*; D004298:Dopamine / Q000134:cerebrospinal fluid; D006719:Homovanillic Acid / Q000134:cerebrospinal fluid; D006897:Hydroxyindoleacetic Acid / Q000134:cerebrospinal fluid; D008297:Male; D008781:Methyltyrosines / Q000134:cerebrospinal fluid; D009638:Norepinephrine / Q000134:cerebrospinal fluid; D051381:Rats; D011919:Rats, Inbred Strains; D012701:Serotonin / Q000134:cerebrospinal fluid; D014364:Tryptophan / Q000134:cerebrospinal fluid*; D014443:Tyrosine / Q000031:analogs & derivatives*
D008781:Methyltyrosines; D015102:3,4-Dihydroxyphenylacetic Acid; D012701:Serotonin; D014443:Tyrosine; D006897:Hydroxyindoleacetic Acid; D004295:Dihydroxyphenylalanine; D014364:Tryptophan; D006916:5-Hydroxytryptophan; C008404:3-methoxytyrosine; D004298:Dopamine; D009638:Norepinephrine; D006719:Homovanillic Acid