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

doi:10.3969/j.issn.1673-8640.2014.01.017

Abstract

Abstract: 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-N-hydroxysuccinimidyl 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. Results RP-HPLC with fluorescence detector is quick, reliable and sensitive to measure the amino acid neurotransmitters in hippocampus of rats.

Key words: Amino acid, Neurotransmitter, High performance liquid chromatography, Pre-column derivatization

CLC Number:

Q517

BAI Yu, ZHANG Li, MA Xiaoli, KONG Bin, LI Xinxia, LI Xinyu.. The determination of 5 amino acid neurotransmitters in hippocampus of rats by RP-HPLC with fluorescence detector[J]. , 2014, 29(1): 69-72.

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