Abstract
A method was developed for simultaneous determination of endogenous and newly synthesized neurotransmitter amino acids (4-aminobutyric acid, glutamate and aspartate) and glutamine in brain in vitro. Brain slices were incubated in artificial cerebrospinal fluid in the presence of 13C-labeled precursors (glucose, pyruvate or acetate). After the incubation, the slices were homogenized in cold 80% ethanol and the supernatants were evaporated to dryness. The resultant residues were derivatized with N-methyl-N-( tert.-butyldimethylsilyl)trifluoroacetamide and analyzed by capillary gas chromatography-mass spectrometry in the electron-impact mode. N(O)- tert.-Butyldimethylsilyl derivatives of the naturally occurring amino acids, their 13C-enriched counterparts and deuterated internal standards were detected as their [M—57] + fragments using selected-ion monitoring. The method was shown applicable to studying compartmentation of neurotransmitter amino acids.
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