Rapid and selective determination of neurotransmitter amino acids in rat and human brain is accomplished by pre-column derivatization with o-phthaldehyde-3-mercaptopropionic acid, reversed-phase high-performance liquid chromatographic separation, and fluorimetric detection. Aspartate, taurine, glutamate, and glycine are determined in less than 12 min with intra- and inter-assay precisions of 2.1-9.5% and 4.2-9.2%, respectively. gamma-Aminobutyric acid is determined in less than 8 min with intra- and inter-assay precisions of 1.6 and 11.7%, respectively. Both assays utilize internal standards and require a minimum of sample preparation.