The determination of 5 amino acid neurotransmitters in hippocampus of rats by RP-HPLC with fluorescence detector

Objective To establish a method of reverse phase high performance liquid chromatography( RP-HPLC) with fluorescence detector to determine 5 amino acid [aspartate( Asp),glutamate( Glu),asparagine( Aspa), glutamine( Gln) and glycine( Gly) ]neurotransmitters. Methods The amino acid neurotransmitters in hippocampus of rats were collected by methanol: water = 1 ∶ 1. The RP-HPLC with fluorescence detector with 6-aminoquinolyl-Nhydroxysuccinimidyl carbamate( AQC) pre-column derivatization was used to measure the 5 amino acid neurotransmitters in hippocampus of rats. Results The 5 amino acid neurotransmitters were well and completely separated using elution within 15 min. There was a good linear correlation in the range of 0. 5-100. 0 μg / μL,and the correlation coefficient( r) was≥0. 999 90. The detection limits of Asp,Glu,Aspa,Gln and Gly were 0. 05,0. 068, 0. 037,0. 05 and 0. 02 μg / μL. The precisions [relative standard deviation( RSD) ]were 0. 4%-2. 5%. The recoveries of added amino acids were 70. 27%-128. 20%( RSD 5%). The contents of Asp,Glu,Aspa,Gln and Gly in hippocampus of normal rats were 38. 98 ± 9. 66,126. 42 ± 34. 06,216. 00 ± 0. 75,90. 44 ± 33. 75 and( 12. 95 ± 4. 42) μg / g. Conclusions RP-HPLC with fluorescence detector is quick,reliable and sensitive to measure the amino acid neurotransmitters in hippocampus of rats.