Reverse Phase High Performance Liquid Chromatography for the Determination of Four Amino Acid Neurotransmitters in Rat Brain Tissues

Objective:To develop a high performance liquid chromatography(HPLC) method with fluorescence detection for simultaneous separation and rapid determination of four amino acid neurotransmitters in rat brain tissues,aspartate(Asp),glutamate(Glu),taurine(Tau) and γ-aminobutyric acid(γ-GABA).Methods:The HPLC column,Dikma Inertsil ODS(250 mm×4.6 mm,3.5 μm) was protected with a security guard cartridge(Dikma,C18,4.0 mm×3.0 mm).The precolumn derivatization reagent was o-phthalaldehyde(OPA),and homoserine was used as the internal standard.The mobile phase was a mixture of potassium dihydrogen phosphate buffer(0.1 mol/L,pH 6.0)∶methanol∶acetonitrile(6∶3∶1) with a flow rate of 1 mL/min.Results: Four amino acids were separated within 15 min.A good linear correlation was obtained in the range of 0.1-2 μmol /L amino acid concentration with good extracted recoveries.Conclusions: The established HPLC method is rapid,accurate and sensitive.It is suitable for the detection of neurotransmitter levels in rat brain tissues.