Premium
Simultaneous measurement of dopamine, serotonin, their metabolites and tryptophan in mouse brain homogenates by high‐performance liquid chromatography with dual coulometric detection
Author(s) -
Alvarez JeanClaude,
Bothua Dominique,
Collig Isabelle,
Advenier Charles,
SpreuxVaroquaux Odile
Publication year - 1999
Publication title -
biomedical chromatography
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.4
H-Index - 65
eISSN - 1099-0801
pISSN - 0269-3879
DOI - 10.1002/(sici)1099-0801(199906)13:4<293::aid-bmc863>3.0.co;2-r
Subject(s) - chemistry , chromatography , ascorbic acid , homovanillic acid , high performance liquid chromatography , tryptophan , dopamine , analyte , serotonin , coulometry , detection limit , metabolite , biochemistry , amino acid , medicine , receptor , food science , electrode , electrochemistry
A new rapid and sensitive high‐performance liquid chromatography (HPLC) method for the simultaneous determination of dopamine, 3,4‐dihydroxyphenylacetic acid (DOPAC), 3‐methoxytyramine, 5‐hydroxytryptamine (serotonin), 5‐hydroxyindoleacetic acid (5‐HIAA), homovanillic acid and tryptophan has been developed and applied to mouse frontal cortex, caudate nucleus and dorsal raphe assays. A dual coulometric detector was used with detection at +0.25 and +0.55 V, which allowed the determination of tryptophan. Detection limits for all compounds (0.8–9.0 pg per injection, depending on the compounds) were useful for this application. Owing to great sensitivity of the method, the brain tissue samples can be very small, less than 2 mg. Linearity of standards was excellent ( r > 0.999 in all cases). Intraday and interday precisions for samples analytes were generally acceptable (intraday assay CV < 8.7% and interday assay CV < 7.0% except for DOPAC and 5‐HIAA, which was 11.4% for the low concentrations). Average recoveries of standard additions to sample analytes were >90%. Attention was paid to stability of standard and sample analytes when stored at +4°C or at −70°C with two different homogenizing agents (0.1 M HClO 4 with 10 −7 M ascorbic acid and 0.05 M HClO 4 without ascorbic acid). This simple, rapid and efficient method can be used as a basic research tool for modification of brain neurotransmitters in experimental pharmacological protocols for following psychotropic drug treatments in animals. Copyright © 1999 John Wiley & Sons, Ltd.