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 language | English |
|---|---|
| Pages (from-to) | 149-169 |
| Number of pages | 21 |
| Journal | Analytical Biochemistry |
| Volume | 64 |
| Issue number | 1 |
| DOIs | |
| State | Published - 1975 |
Fingerprint
ASJC Scopus subject areas
- Biochemistry
- Biophysics
- Molecular Biology
Cite this
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. / Smith, J. E.; Lane, J. D.; Shea, P. A.; McBride, W. J.; Aprison, M. H.
In: Analytical Biochemistry, Vol. 64, No. 1, 1975, p. 149-169.Research output: Contribution to journal › Article
}
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 -