Simultaneous determination of glutamate,aspartate,γ-aminobutyric acid and glycine in human serum by RP-HPLC

[Objective] To develop a high performance liquid chromatography(HPLC) for simultaneous separation and rapid determination of four amino acid [glutamate(Glu),aspartate(Asp),γ-aminobutyric acid(GABA),glycine(Gly) ] in human serum.[Methods] The precolumn derivazation reagent was OPA.Derivatives were separated on a Nova-Pak C18(3.9 mm×150 mm,4 μm).Phosphate buffer solution,tetrahydrofuran and methanol as mobile phases,and gradient elution lasted 20 min with fluorescence detection.[Results] The range of calibration curve was(4~400)nmol/mL.The calibrations were characterized by a coefficient of correlation higher than 0.997.The average extracted recovery was 90.0%~93.1%,RSD%9.87%.[Conclusion] Our results showed that the method was rapid,accurate and simple.