A method for concurrent measurement of picomole quantities of acetylcholine, choline, dopamine, norepinephrine, serotonin, 5-hydroxytryptophan, 5-hydroxyindoleacetic acid, tryptophan, tyrosine, glycine, aspartate, glutamate, alanine, and gamma-aminobutyric acid in single tissue samples from different areas of rat central nervous system

J. E. Smith, J. D. Lane, P. A. Shea, W. J. McBride, M. H. Aprison

Research output: Contribution to journalArticle

132 Citations (Scopus)

Abstract

A rapid and sensitive method for separation and concurrent assay of 14 compounds at the picomole level in individual rat brain parts is described. The putative amino acid neurotransmitters (aspartate, γ-aminobutyric acid, glutamate, and glycine) are extracted from a 20-30 mg portion of the tissue with 5% TCA and assayed as their respective DNP-amino acid methyl ester derivatives by glc. Four other putative neurotransmitters (acetylcholine, dopamine, norepinephrine, and serotonin) and some of their precursors and metabolites (choline, tryptophan, 5-hydroxytryptophan, 5-hydroxyindoleacetic acid, and tyrosine) are extracted in 1 n formic acid-acetone ( v v: 15 85) from the remaining tissue. The lipids are removed with a heptane-chloroform ( v v: 8 1) wash and the aqueous phase is dried at 37°C under N2. The dried extract is dissolved in water (pH 4). With one portion of this solution, acetylcholine and choline are assayed using a radioenzymatic method whereas with the rest, dopamine, norepinephrine, serotonin, 5-hydroxyindoleacetic acid, 5-hydroxytryptophan, tryptophan, and tyrosine are separated with three ion exchange resins arranged in tandem. These compounds are assayed fluorometrically with modified microadaptations of previously reported methods.

Original languageEnglish
Pages (from-to)149-169
Number of pages21
JournalAnalytical Biochemistry
Volume64
Issue number1
DOIs
StatePublished - 1975

Fingerprint

5-Hydroxytryptophan
Hydroxyindoleacetic Acid
formic acid
Neurology
Choline
Aspartic Acid
Tryptophan
Alanine
Glycine
gamma-Aminobutyric Acid
Acetylcholine
Neurotransmitter Agents
Tyrosine
Rats
Glutamic Acid
Dopamine
Serotonin
Norepinephrine
Central Nervous System
Tissue

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

@article{9e7268c84828415596e7145d86f04033,
title = "A method for concurrent measurement of picomole quantities of acetylcholine, choline, dopamine, norepinephrine, serotonin, 5-hydroxytryptophan, 5-hydroxyindoleacetic acid, tryptophan, tyrosine, glycine, aspartate, glutamate, alanine, and gamma-aminobutyric acid in single tissue samples from different areas of rat central nervous system",
abstract = "A rapid and sensitive method for separation and concurrent assay of 14 compounds at the picomole level in individual rat brain parts is described. The putative amino acid neurotransmitters (aspartate, γ-aminobutyric acid, glutamate, and glycine) are extracted from a 20-30 mg portion of the tissue with 5{\%} TCA and assayed as their respective DNP-amino acid methyl ester derivatives by glc. Four other putative neurotransmitters (acetylcholine, dopamine, norepinephrine, and serotonin) and some of their precursors and metabolites (choline, tryptophan, 5-hydroxytryptophan, 5-hydroxyindoleacetic acid, and tyrosine) are extracted in 1 n formic acid-acetone ( v v: 15 85) from the remaining tissue. The lipids are removed with a heptane-chloroform ( v v: 8 1) wash and the aqueous phase is dried at 37°C under N2. The dried extract is dissolved in water (pH 4). With one portion of this solution, acetylcholine and choline are assayed using a radioenzymatic method whereas with the rest, dopamine, norepinephrine, serotonin, 5-hydroxyindoleacetic acid, 5-hydroxytryptophan, tryptophan, and tyrosine are separated with three ion exchange resins arranged in tandem. These compounds are assayed fluorometrically with modified microadaptations of previously reported methods.",
author = "Smith, {J. E.} and Lane, {J. D.} and Shea, {P. A.} and McBride, {W. J.} and Aprison, {M. H.}",
year = "1975",
doi = "10.1016/0003-2697(75)90417-0",
language = "English",
volume = "64",
pages = "149--169",
journal = "Analytical Biochemistry",
issn = "0003-2697",
publisher = "Academic Press Inc.",
number = "1",

}

TY - JOUR

T1 - A method for concurrent measurement of picomole quantities of acetylcholine, choline, dopamine, norepinephrine, serotonin, 5-hydroxytryptophan, 5-hydroxyindoleacetic acid, tryptophan, tyrosine, glycine, aspartate, glutamate, alanine, and gamma-aminobutyric acid in single tissue samples from different areas of rat central nervous system

AU - Smith, J. E.

AU - Lane, J. D.

AU - Shea, P. A.

AU - McBride, W. J.

AU - Aprison, M. H.

PY - 1975

Y1 - 1975

N2 - A rapid and sensitive method for separation and concurrent assay of 14 compounds at the picomole level in individual rat brain parts is described. The putative amino acid neurotransmitters (aspartate, γ-aminobutyric acid, glutamate, and glycine) are extracted from a 20-30 mg portion of the tissue with 5% TCA and assayed as their respective DNP-amino acid methyl ester derivatives by glc. Four other putative neurotransmitters (acetylcholine, dopamine, norepinephrine, and serotonin) and some of their precursors and metabolites (choline, tryptophan, 5-hydroxytryptophan, 5-hydroxyindoleacetic acid, and tyrosine) are extracted in 1 n formic acid-acetone ( v v: 15 85) from the remaining tissue. The lipids are removed with a heptane-chloroform ( v v: 8 1) wash and the aqueous phase is dried at 37°C under N2. The dried extract is dissolved in water (pH 4). With one portion of this solution, acetylcholine and choline are assayed using a radioenzymatic method whereas with the rest, dopamine, norepinephrine, serotonin, 5-hydroxyindoleacetic acid, 5-hydroxytryptophan, tryptophan, and tyrosine are separated with three ion exchange resins arranged in tandem. These compounds are assayed fluorometrically with modified microadaptations of previously reported methods.

AB - A rapid and sensitive method for separation and concurrent assay of 14 compounds at the picomole level in individual rat brain parts is described. The putative amino acid neurotransmitters (aspartate, γ-aminobutyric acid, glutamate, and glycine) are extracted from a 20-30 mg portion of the tissue with 5% TCA and assayed as their respective DNP-amino acid methyl ester derivatives by glc. Four other putative neurotransmitters (acetylcholine, dopamine, norepinephrine, and serotonin) and some of their precursors and metabolites (choline, tryptophan, 5-hydroxytryptophan, 5-hydroxyindoleacetic acid, and tyrosine) are extracted in 1 n formic acid-acetone ( v v: 15 85) from the remaining tissue. The lipids are removed with a heptane-chloroform ( v v: 8 1) wash and the aqueous phase is dried at 37°C under N2. The dried extract is dissolved in water (pH 4). With one portion of this solution, acetylcholine and choline are assayed using a radioenzymatic method whereas with the rest, dopamine, norepinephrine, serotonin, 5-hydroxyindoleacetic acid, 5-hydroxytryptophan, tryptophan, and tyrosine are separated with three ion exchange resins arranged in tandem. These compounds are assayed fluorometrically with modified microadaptations of previously reported methods.

UR - http://www.scopus.com/inward/record.url?scp=0016664248&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0016664248&partnerID=8YFLogxK

U2 - 10.1016/0003-2697(75)90417-0

DO - 10.1016/0003-2697(75)90417-0

M3 - Article

C2 - 237425

AN - SCOPUS:0016664248

VL - 64

SP - 149

EP - 169

JO - Analytical Biochemistry

JF - Analytical Biochemistry

SN - 0003-2697

IS - 1

ER -