RP-LC with Fluorescence Detection of Amino Acids in Rat Brain Synaptosomes

Abstract

For the first time, a simple and validated reversed-phase liquid chromatography (RP-LC) with fluorescence detection has been developed for the simultaneous analysis of glutamate (Glu), c-aminobutyric acid (GABA), glycine (Gly) and taurine (Tau) in Wistar and tremor rats brain synaptosomes. The samples were separated on a C18 analytical column with gradient elution of methanol and 0.1 mol L -1 potassium acetate at a flow rate of 1 mL min -1 . Total run time was approximately 25 min. All calibration curves exhibited good linearity (r 2 [ 0.999) within test ranges. The reproducibility was estimated by intra-and inter-day assays and RSD values were less than 2.48%. The recoveries were between 96.32 and 105.21%. The method was successfully applied to the quantification of amino acids in Wistar and tremor rats brain synaptosomes. Through this developed protocol, the levels of Glu in hippocampal and prefrontal cortical synaptosomes of tremor rats were both significantly elevated than those of adult Wistar rats whereas significantly decreased concentrations of GABA and Gly were observed in the hippocampal region of tremor rats without evident difference in the prefrontal cortex between experimental and control groups. In addition, our studies also showed a marked elevation of Tau in tremor rats hippocampal synaptosomes although there was no pronounced difference in the prefrontal cortical region of Wistar and tremor rats.

Aberrant changes of somatostatin and neuropeptide Y in brain of a genetic rat model for epilepsy: tremor rat

“…Moreover, this peptide displays enhanced expression in different brain areas from varied epileptic animal models (Goto et al 2010, Kokaia 2011, which are coincide with our results in this study, suggesting that the TRM epileptiform activity might not only make an effect upon SST system, but also influence NPY's expression or secretion. Due to the diminished GABA and elevated glutamine reported in our previous study (Mao et al 2011b), the increased amount of SST or NPY could be recognized as a self-neuroprotection against the aberrant neurotransmission circuitry in TRM brains. Although these neuropeptides are likely to take part in TRM self-defense process, the epileptogenesis in current epileptic animals do not dampened totally.…”

Study of the rapid detection of γ-aminobutyric acid in rice wine based on chemometrics using near infrared spectroscopy

“…In recent years, it has been reported that an amino acid analyser, GC or LC-MS, column chromatography fluorescence assay and high performance liquid chromato-graphy (HPLC) have all been applied to the detection of γ-aminobutyric acid (Constantinos et al 2004;Zhang and Alan 1997;Zhang and Sun 2005;Mao et al 2011;Osborne et al 1999;Silva et al 2009;Ryutaro et al 2012); however, the detection of GABA using the near infrared spectroscopy (NIR) method, employing chemometrics, has not previously been published.…”