Quantification of N-acetylaspartylglutamate, an N-terminal blocked dipeptide neurotransmitter candidate, in brain slice superfusates by gas chromatography-mass spectrometry.

N-Acetylaspartylglutamate is a highly promising neurotransmitter candidate. A method for its quantification has been developed that allows to investigate its stimulation-induced release from brain slices. The method is based on ion-exchange prepurification, derivatization with HCl in methanol, separation by capillary gas chromatography and quantification by ammonia chemical ionization mass spectrometry with selected-ion monitoring. Deuterium-labelled N-acetylaspartylglutamate is used as internal standard. The method has been validated, also with respect to possible interfering compounds. A limit of quantification in the low pmol to high fmol range has been achieved, which is clearly sufficient for the intended purpose. A detailed analytical procedure is given, and alternatives for some of the different steps are discussed. Derivatization with diazomethane instead of methanolic HCl turned out to be impracticable. The method may well be applicable to certain other N-terminal blocked di- and tripeptides and to acylated amino acids.