Determination of amino acid neurotransmitters in the hippocampal slices perfusate by RP HPLC

AIM To establish a method for simultaneous determination of trace amino acid neurotransmitters Glu, Asp, Tau and GABA by HPLC was improved. METHODS The sample prepared by rat hippocampal slices perfusation was lyophi lized and resoluted, 4 kinds of trace amino acids were determinated by HPLC which involved pre column derivatization with o phthalaldehyde, reversed phase gradient elution and fluorescence detection. RESULTS A good linear relationship existed between peak area and concentration of amino acids. The correlation coefficients ( r ) were: Asp 0.9920 , Glu 0.9667, Tau 0.9803 and GABA 0.9584 respectively. The precisions (%) were Glu 4.3, Asp 4.6, Tau 6.7 and GABA 7.1 for interassay CV. The detection limits ( pmol· L -1 ) were: Asp 0.17, Glu 0.35, Tau 0.68, GABA 1.12 and the recovery rates of 4 amino acids (%) were Asp 106±7, Glu 101±5, Tau 94±6 and GABA 92±11 respectively. CONCLUSION The operation of the derivatization is easy, the mobile phase is cheap and this method has sufficient sensitivity, specificity and less separation time. Also it is good for determination of amino acid in other biologic sample.