Determination of neurotransmitter amino acids in mouse central nervous system by CE–LIF

Abstract

An MEKC method with LIF detection has been developed for the determination of seven neurotransmitter amino acids (NAAs) using 1,3,5,7-tetramethyl-8-(N-hydroxysuccinimidyl butyric ester)difluoroboradiaza-S-indacene as the labeling reagent. After derivatization at room temperature for 30 min, the seven target NAAs including glycine, alanine, γ-aminobutyric acid, taurine, glutamine, glutamic acid, and aspartic acid were separated in running buffer, which consisted of 70 mM pH 4.00 H3 PO4 /Na3 PO4 buffer, 5.5 mM cetyltrimethyl ammonium bromide and 20% v/v acetonitrile within 17 min. The LODs were 2 ~ 14 × 10(-10) M without interference from other coexisting amino acids. The proposed method has been applied to the analysis of NAAs in the central nervous systems of healthy mice and those with Alzheimer's disease with recoveries of 92-104%.

Analytical Techniques in Neuroscience: Recent Advances in Imaging, Separation, and Electrochemical Methods

“…After derivatization with TMBB-Su, glycine, alanine, γ-aminobutyric acid, taurine, glutamine, glutamic acid, and aspartic acid were cleanly separated using a cationic surfactant with MEKC. 106 A primary advantage of CE separations is their speed, and CE-LIF separations can be performed on the 10 second time frame. For example, salamander retinas were prepared and dopamine, D-serine, L-serine, phosphoethanolamine, glutathione, 2-aminoadipic acid (internal standard), glutamate, and L-aspartate were separated using microdialysis with an online derivatization for CE-LIF.…”

“…One new fluorophore that helped combat this obstacle was 4-fluoro-7-nitrobenzo-2-oxa-1,3-diazole (NBD-F), which had a highly efficient reaction time of 5 min with primary and secondary amines . Other fluorophores were designed to improve the sensitivity of a specific analyte including 2,4,6-trinitrobenzenesulfonic acid (TNBS) for the derivatization of adenosine and inosine or 1,3,5,7-tetramethyul-8-( N -hydroxysuccinimidyl butyric ester) difluoroboradiaza- S -indicene (TMBB-Su) for neurotransmitter amino acids. , Micellar electrokinetic chromatography (MEKC) is a form of CE that is frequently used because a surfactant is added to separate neutral molecules. After derivatization with TMBB-Su, glycine, alanine, γ-aminobutyric acid, taurine, glutamine, glutamic acid, and aspartic acid were cleanly separated using a cationic surfactant with MEKC .…”

Recent advances in amino acid analysis by capillary electromigration methods, 2013–2015

“…The concentrations of Ala, Gly, Glu, and Asp are higher in the brain cortex of healthy mice. In the transgenic ones, concentrations of GABA, Gln, and Taurine are higher in the brain cortex, while they are all at the highest concentration in the lumbar spinal cord .…”