Rapid method for micro-analysis of endogenous amino acid neurotransmitters in brain perfusates in the rat by isocratic HPLC-EC.
A simplified method is described for the isocratic analysis of endogenous amino acid neurotransmitters contained in brain perfusates by high performance liquid chromatography (HPLC) with electrochemical detection (EC). Pre-column o-phthalaldehyde (OPA) tert-butylthiol derivatives of the amino acids were injected into a C18 3 microns column. After linear concentration curves for standard solutions were obtained, the content of 6 amino acid neurotransmitters was analyzed in push-pull perfusates obtained from the hypothalamus and cerebral cortex of the unrestrained rat. Each analysis which included the simultaneous quantification of aspartic acid, glutamic acid, glutamine, glycine, taurine and gamma-aminobutyric acid (GABA), was completed in less than 15 min. The sensitivity of the assay ranged from 1.0 to 5.0 pmol of each amino acid contained within a 20 microliters aliquot of each perfusion sample.
D000596:Amino Acids / Q000032:analysis*; D000818:Animals; D001923:Brain Chemistry; D002540:Cerebral Cortex / Q000032:analysis; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004563:Electrochemistry / Q000379:methods*; D005971:Glutamates / Q000032:analysis; D018698:Glutamic Acid; D007031:Hypothalamus / Q000032:analysis; D018377:Neurotransmitter Agents / Q000032:analysis*; D051381:Rats; D013654:Taurine / Q000032:analysis; D005680:gamma-Aminobutyric Acid / Q000032:analysis
D000596:Amino Acids; D005971:Glutamates; D018377:Neurotransmitter Agents; D013654:Taurine; D018698:Glutamic Acid; D005680:gamma-Aminobutyric Acid