Summary
For simultaneous assay of the five neurotransmitter amino acids, Asp, Glu, Gly, Tau, and GABA in brain tissues, a very rapid and simple chromatographic method using high-performance liquid chromatography with electrochemical detection in combination with o-phthalaldehyde derivatization is described. Because the present method permits the determination of these five amino acids within less than five minutes in one chromatographic run, up to 100 samples a working day can be analyzed using an autosampler. Withinrun coefficients of variation for these five amino acids were less than 2% (n=20). The quantitative detection limit was 2.5 pmol for the 5 amino acids. The present method has been applied to the measurement of the five amino acid neurotransmitter levels in several discrete brain regions of mice treated with and without electroconvulsive shock.
Similar content being viewed by others
References
Allison LA, Mayer GS, Shoup RE (1984) o-Phthalaldehyde derivatives of amines for high-speed liquid chromatography/electrochemistry. Anal Chem 56: 1089–1096
Balcom GJ, Lenox RH, Meyerhoff JL (1975) Regional γ-aminobutyric acid levels in rat brain determined after microwave fixation. J Neurochem 24: 609–613
Bowdler JM, Green AR (1982) Regional rat brain benzodiazepine receptor number and γ-aminobutyric acid concentration following a convulsion. Br J Pharmacol 76: 291–298
Chapman AG, Hart GP (1988) Anticonvulsant drug action and regional neurotransmitter amino acid changes. J Neural Transm 72: 201–212
Chapman AG, Cheetham SC, Hart GP, Meldrum BS, Westerberg E (1985) Effects of two convulsant β-carboline derivatives, DMCM and β-CCM, on regional neurotransmitter amino acid levels and on in vitro D-[3 H]-aspartate release in rodents. J Neurochem 45: 370–381
Chapman AG, Halsey MJ, Hart GP, Luff NP, Meldrum BS, Wardley-Smith B (1986) Regional amino acid concentration in the brains of rats exposed to high pressures. J Neurochem 47: 314–317
Cordoba F, Yusta B, Munoz-Blanco J (1984) Changes in neurotransmitter amino acids and protein in CNS areas of mice subjected to differential housing conditions. Pharmacol Biochem Behav 21: 349–352
Donzanti BA, Yamamoto BK (1988) An improved and rapid HPLC-EC method for the isocratic separation of amino acid neurotransmitters from brain tissue and microdialysis perfusates. Life Sci 43: 913–922
Fagg GE, Foster AC (1983) Amino acid neurotransmitters and their pathways in the mammalian central nervous system. Neuroscience 9: 701–719
Farrant M, Zia-Gharib F, Webster RA (1987) Automated pre-column derivatization with o-phthalaldehyde for the determination of neurotransmitter amino acids using reversedphase liquid chromatography. J Chromatogr 417: 385–390
Fletcher A, Fowler LJ (1982) A regional study of 4-aminobutyrate metabolism and amino acid levels in rat brain following chronic oral administration of ethanolamine o-sulfate. J Neurochem 38: 1022–1028
Glowinski J, Iversen LL (1966) Regional studies of catecholamines in the rat brain. I. The disposition of [3 H]-norepinephrine, [3 H]-dopamine and [3 H]-dopa in various regions of the brain. J Neurochem 13: 655–669
Green AR, Metz A, Minchin MCW, Vincent ND (1987) Inhibition of the rate of GABA synthesis in regions of rat brain following a convulsion. Br J Pharmacol 92: 5–11
Jakubovic A, Fu D, Fibiger HC (1987) Factors affecting the stability and separation of biogenic amines and their metabolites. J Pharmacol Methods 17: 1–14
Krstulovic AM (1982) Investigations of catecholamine metabolism using high-performance liquid chromatography; analytical methodology and clinical applications. J Chromatogr 229: 1–34
Murai S, Miyate H, Saito H, Nagahama H, Masuda Y, Itoh T (1988) Simple determination of acetylcholine and choline within 4 min by HPLC-ECD and immobilized enzyme column in mice brain areas. J Pharmacol Methods 21: 255–262
Murai S, Saito H, Masuda Y, Itoh T (1990) Very rapid and simple assay of taurine in the brain within two minutes by high-performance liquid chromatography with electrochemical detection. J Pharmacol Methods 23: 195–202
Peinado JM, McManus KT, Myers RD (1986) Rapid method for micro-analysis of endogenous amino acid neurotransmitters in brain perfusates in the rat by isocratic HPLC-EC. J Neurosci Methods 18: 269–276
Saller CF, Czupryna MJ (1989) γ-Aminobutyric acid, glutamate, glycine and taurine analysis using reversed-phase high-performance liquid chromatography and ultraviolet detection of dansyl chloride derivatives. J Chromatogr 487: 167–172
Westerberg E, Chapman AG, Meldrum BS (1983) Effect of 2-amino-7-phosphonoheptanoic acid on regional brain amino acid levels in fed and fasted rodents. J Neurochem 41: 1755–1760
Zecca L, Ferrario P (1985) Determination of neurotransmitter amino acids by high-performance liquid chromatography with fluorescence detection. J Chromatogr 337: 391–396
Zielke HR (1985) Determination of amino acids in the brain by high-performance liquid chromatography with isocratic elution and electrochemical detection. J Chromatogr 347: 320–324
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Murai, S., Saito, H., Abe, E. et al. A rapid assay for neurotransmitter amino acids, aspartate, glutamate, glycine, taurine and γ-aminobutyric acid in the brain by high-performance liquid chromatography with electrochemical detection. J. Neural Transmission 87, 145–153 (1992). https://doi.org/10.1007/BF01245016
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF01245016